comparative evaluation of the 3m petrifilm aqua plate ... · page 1 an independent laboratory...

An independent laboratory comparative evaluation of the 3M™ Petrifilm™ Aqua Plate method (3M, St. Paul, Minnesota) for quantitation

of heterotrophic bacteria, coliforms, Enterobacteriaceae, Escherichia coli, and yeast and mold in four types of bottled water (Purified,

Regional Spring, Natural Spring, and Carbonated Natural Spring) was conducted at Q Laboratories, Inc., Cincinnati, OH. The 3M

Petrifilm Aqua Heterotrophic Count Plate (AQHC) was compared to the Standard Methods for the Examination of Water and Wastewater

(SMEWW) and the ISO reference method for the quantitation of heterotrophic bacteria through direct plating and membrane

filtration. The 3M Petrifilm Aqua Coliform Count Plate (AQCC) and 3M Petrifilm E. Coli/Coliform Count Plate (EC) were compared to

the SMEWW and the ISO method for the enumeration of coliforms and E. coli through membrane filtration. The 3M Petrifilm Aqua

Enterobacteriaceae Count Plate (AQEB) was compared to a customer method for the enumeration of Enterobacteriaceae through

membrane filtration. The 3M Petrifilm Aqua Yeast and Mold Count Plate (AQYM) was compared to the SMEWW and a customer

method for the enumeration of yeast and mold through membrane filtration. Analysis of variance showed that there was no statistical

difference between the 3M Petrifilm Aqua Plate methods at a low and a high inoculation level and their respective reference methods,

except the mean log count per filter for the 3M Petrifilm Aqua AQCC Plate method was greater in value when compared to the SMEWW

reference agar method at the high inoculation level and the day 5 mean log yeast and mold count per filter was greater in value

between the 3M Petrifilm Aqua AQYM Plate method and a customer method at the high inoculation level.

This report presents the analytical results for comparison of the 3M™ Petrifilm™ Aqua Plate methods to reference methods and

customer methods for the quantitation of heterotrophic bacteria, coliforms, Enterobacteriaceae, Escherichia coli, and yeast and

molds in bottled water. All analyses were conducted at Q Laboratories, Inc. (Cincinnati, OH). All 3M Petrifilm Plates were

provided by 3M.

Materials and Methods

The methodology for this study was followed as outlined in 3M’s Protocol: 3M Petrifilm Plate Project Claims Verification Study

Protocol Version 5.0 (October 8, 2010). The study consisted of quantifying heterotrophic bacteria, coliforms, Enterobacteriaceae,

Escherichia coli, and yeast and mold on various 3M Petrifilm Aqua Plates and reference and customer method agars. Two separate

lots of four types of bottled water (Purified, Regional Spring, Natural Spring, and Carbonated Natural Spring) were analyzed

by each plate method. All Natural Spring and Carbonated Natural Spring waters were analyzed for the presence of naturally

occurring bacteria. Due to the low level of naturally occurring bacteria found in those samples, two lots of each of the four types

of bottled water were filter sterilized and inoculated with one of three organisms specified for each plating method as outlined

Q Laboratories, Inc. is an independent company and is not affiliated with, nor a subsidiary of 3M. Q Laboratories, Inc. was contracted to perform an independent study by and for 3M and the data generated by Q Laboratories, Inc. during the execution of the study does not represent an endorsement of any product(s) by Q Laboratories, Inc.

Comparative Evaluation of the 3M™ Petrifilm™ Aqua Plate Method vs. Multiple Reference Methods in Bottled Water —

Claims Verification Study

Erin Crowley, Patrick Bird, Kiel Fisher, Katherine Goetz, Megan Boyle, M. Joseph Benzinger Jr., Mark Juenger, Shana Gibbs, James R. Agin, David G. Goins

Q Laboratories, Inc. 1400 Harrison Avenue Cincinnati, OH 45214

in Table 1 in Appendix 1. Water samples inoculated with organisms for heterotrophic bacteria, coliforms, Enterobacteriaceae,

and E. coli were allowed to equilibrate for two weeks at room temperature (24 ± 2°C) and analyzed following the 3M Petrifilm

Aqua Plate, reference and customer methods. Water samples inoculated with yeast and molds were held for 24 hours at room

temperature (24 ± 2°C) and then analyzed following the 3M Petrifilm Aqua Plate, reference and customer methods. All products

for this assay were purchased prior to study initiation from various local supermarkets.

Table A: Waters Used In Evaluation

Water Type Brand Country of Manufacture

Purified

Nestle Pure Life United States

Aquafina United States

Dasani United States

Regional Spring

Trauth Dairy United States

Ice Mountain United States

Kroger United States

Natural Spring

Evian France

Jana Croatia

Fiji Fiji

Ty Nant Wales

Voss Flat Norway

Natural Spring (Carbonated)

Voss Sparkling Norway

Gerolsteiner Germany

Apollinaris Germany

Perrier France

Heterotrophic Plate Count For the analysis of heterotrophic bacteria, two lots each of the Natural Spring Waters and Carbonated Natural Spring Waters

listed in Table A were screened for natural occurring bacteria in duplicate by direct plate count. The background screen revealed

no natural contamination in the Carbonated Natural Spring waters and very low levels of natural contamination in the Natural

Spring Waters. Therefore, in addition to testing uninoculated Natural Spring Waters by the methods outlined below, both the

Natural Spring Waters and the Carbonated Natural Spring Waters were filter-sterilized with an 0.22μm filter and inoculated,

along with the Regional Spring and Purified waters listed in Table A, with one of three organisms (Escherichia coli ATCC

#25922, Enterococcus faecium ATCC #19434, and Pseudomonas aeruginosa ATCC #15442) as specified in Table 1 of

Appendix 1.

A. Direct Plating Comparisons.

For uninoculated samples, Natural Spring Waters were analyzed by the SMEWW method and the ISO method outlined below.

In order to achieve a countable range, ten-fold serial dilutions were made using filter sterilized (22 μm) dilution blanks of each

brand of water.

For inoculated waters, separate samples were spiked at a low level inoculum targeting 25 CFU/mL and a high level inoculum

targeting 75 CFU/mL. After a two-week equilibration, waters were analyzed by the SMEWW method and the ISO method

described below. In order to achieve a countable range, ten-fold serial dilutions were made using filter sterilized (22 μm)

dilution blanks of each brand of water.

Duplicate 1mL samples were plated on the 3M Petrifilm Aqua AQHC Plate and on reference method agar pour plates. One set

of test portions was incubated at 36 ± 1°C for 45–48 hours. The test portions at this incubation condition included Standard

Methods Plate Count Agar (PCA; SMEWW method), Yeast Extract Agar (YEA; ISO method), and 3M Petrifilm Aqua AQHC

Plates. Additionally, another set of test portions was prepared and incubated at 22 ± 2°C for 68 ± 4 hours. Test portions at this

incubation condition included YEA and 3M Petrifilm Aqua AQHC Plates. After incubation, all plates were counted. On the 3M

Petrifilm Aqua AQHC Plate, all red colonies were counted. On PCA and YEA, all colonies in and on the agar were counted.

B. Filtered Sample Comparison.

For uninoculated samples, Natural Spring Waters were analyzed by the SMEWW method outlined below. In order to achieve a

countable range, ten-fold serial dilutions were made using filter sterilized (22μm) dilution blanks of each brand of water.

For inoculated samples, both lots of each of the four water types were inoculated with one of the three organisms outlined

above. Separate samples were spiked at a low level inoculum targeting 25 CFU/100mL and a high level inoculum targeting

75 CFU/100mL. After a two-week equilibration, waters were analyzed by the SMEWW method described below. The 3M

Petrifilm Aqua AQHC Plates used in this portion of the evaluation were pre-hydrated with 1 ml of sterile deionized water, held

at refrigerated temperature for no longer than a week, and allowed to come to room temperature prior to analysis. In order to

achieve a countable range, ten-fold serial dilutions were made using filter sterilized (22μm) dilution blanks of each brand of

water. Samples were analyzed in duplicate by filtering 100mL of sample through Mixed Cellulose Ester (MCE) membrane

filters and placed onto PCA plates and onto 3M Petrifilm Aqua AQHC Plates. All plates were incubated at 35 ± 1°C for 48 ± 3

hours. On the 3M Petrifilm Aqua AQHC Plate, all red colonies were counted. On PCA, all colonies on the filter were counted.

Detailed results are presented in Table 2 of Appendix 1.

Coliform Plate Count

Two lots of all four water types were inoculated with one of three organisms (Escherichia coli ATCC #25922, Klebsiella

pneumonia ATCC #4352, and Enterobacter aerogenes ATCC #35029) as specified in Table 1 of Appendix 1. Samples were

analyzed by membrane filtration by both SMEWW and ISO 9308-1:2000: Water quality – Detection and enumeration of

Escherichia coli and coliform bacteria methods and compared to the 3M Petrifilm Aqua Coliform Count Plate (AQCC) method.

For the SMEWW method comparison, separate samples were spiked at a low level inoculum targeting 25 CFU/100mL and a

high level inoculum targeting 75 CFU/100mL. For the ISO method comparison, separate samples were spiked at a low level

inoculum targeting 25 CFU/250mL and a high level inoculum targeting 75 CFU/250mL. After a two-week equilibration, waters

were analyzed by the methods described below. For both comparisons, the 3M Petrifilm Aqua AQCC Plates were pre-hydrated

with 1 ml of sterile deionized water, held at refrigerated temperature for no longer than a week, and allowed to come to room

temperature prior to analysis. In order to achieve a countable range, ten-fold serial dilutions were made using filter sterilized

(22μm) dilution blanks of each brand of water.

A. SMEWW Method comparison.

For the comparison with the SMEWW method, 100mL portions of inoculated samples were filtered in duplicate through

MCE membrane filters and placed onto 3M Petrifilm Aqua AQCC Plates and endo LES agar plates. 3M Petrifilm Aqua

AQCC Plates were incubated for 24 ± 2 hours at 35 ± 0.5°C and endo LES agar plates were incubated for 22–24 hours at 35

± 0.5°C. After incubation, 3M Petrifilm Aqua AQCC Plates were examined for typical colonies (red colony with associated

gas bubbles) and atypical colonies (red without gas bubbles). The endo LES agar plates were observed for typical colonies that

were pink to dark red with a green metallic surface sheen. One typical colony (both endo LES and 3M Petrifilm Aqua AQCC

Plate) and two atypical colonies (3M Petrifilm Aqua AQCC Plate only) per sample were picked to Lauryl Tryptose Broth

(LTB) tubes and incubated at 35 ± 0.5°C for 48 ± 2 hours. After incubation, LTB tubes were examined for gas production,

and a loopful of broth from positive tubes was transferred to Brilliant Green Lactose Bile (BGLB) tubes and incubated at 35

± 0.5°C for 48 ± 2 hours. After incubation, tubes were observed for gas production. Samples producing gas were recorded as

positive. Atypical colonies that produced reactions typical of coliforms were recorded as coliforms.

B. ISO Method comparison.

Duplicate 250mL portions of inoculated samples were filtered through MCE filters and placed on 3M Petrifilm Aqua AQCC

Plates or through Cellulose Nitrate (CN) membrane filters and placed onto Lactose TTC agar with Tergitol 7 plates. 3M

Petrifilm Aqua AQCC Plates were incubated at 36 ± 2°C for 24 ± 2 hours and Lactose TTC plates were incubated at 36 ±

2°C for 21 ± 3 hours. After incubation 3M Petrifilm Aqua AQCC Plates were examined for typical colonies (red colony with

associated gas bubbles) and atypical colonies (red without gas bubbles). Lactose TTC plates were observed for typical yellow

to yellow-orange colonies with yellow acid zones. One typical colony (both Lactose TTC and 3M Petrifilm Aqua AQCC

Plate) and two atypical colonies (3M Petrifilm Aqua AQCC Plate only) per sample were picked to TSA and incubated at

36 ± 2°C for 24 ± 2 hours. Representative colonies on the TSA were confirmed according to the ISO reference method by

conducting the spot Oxidase test. Additionally, colonies were picked and inoculated into 3mL tryptophan broth. Broth was

incubated at 44 ± 0.5°C for 21 h ± 3 hours, then 0.2–0.3mL of Kovak’s reagent was added to test for the presence of indole.

Samples that developed a cherry-red color at the surface of the broth were recorded as positive for indole production. Those

colonies that produced oxidase negative and indole negative results were recorded as coliforms. Colonies that produced

oxidase negative and indole positive results were recorded as E. coli.

All confirmed (typical and confirmed atypical) counts were used in the data analysis. Detailed results are presented in Table 3

of Appendix 1.

Enterobacteriaceae Plate Count

Two lots of all four types of water samples were inoculated with one of three organisms (Escherichia coli ATCC #25922,

Klebsiella pneumonia ATCC #4352, and Enterobacter aerogenes ATCC #35029) as specified in Table 1 of Appendix 1. For the

comparison, separate samples were spiked at a low level inoculum targeting 25 CFU/100mL and a high level inoculum targeting

75 CFU/100mL. After a two-week equilibration, waters were analyzed by the method described below. The 3M Petrifilm Aqua

AQEB Plates were pre-hydrated with 1mL of sterile deionized water, held at refrigerated temperature for no longer than a week,

and allowed to come to room temperature prior to analysis. In order to achieve a countable range, ten-fold serial dilutions were

made using filter sterilized (22 μm) dilution blanks of each brand of water.

Samples were analyzed by membrane filtration by both a customer method and the 3M Petrifilm Aqua Enterobacteriaceae

Count Plate (AQEB) method. 100mL portions of inoculated samples were filtered through MCE membrane filters and placed

onto 3M Petrifilm Aqua AQEB Plates in duplicate and onto Violet Red Bile Glucose (VRBG) agar in duplicate. 3M Petrifilm

Aqua AQEB Plates were incubated at 36 ± 1°C for 24 ± 2 hours and VRBG plates were incubated at 36 ± 1°C for 24 hours.

After incubation, 3M Petrifilm Aqua AQEB Plates were observed for typical colonies (red colony with gas and/or a yellow acid

zone) and VRBG plates were observed for typical pink to dark purple colonies with a halo precipitation in the surrounding agar.

Detailed results are presented in Table 4 of Appendix 1.

Escherichia coli Plate Count

Two lots of all four types of water samples were inoculated with one of three organisms (Escherichia coli ATCC #25922,

Klebsiella pneumonia ATCC #4352, and Enterobacter aerogenes ATCC #35029) as specified in Table 1 of Appendix 1 and

analyzed by membrane filtration by both the SMEWW and the ISO 9308-1:2000: Water quality – Detection and enumeration

of Escherichia coli and coliform bacteria methods and the 3M Petrifilm E. Coli/Coliform Count Plate (EC) method. For the

SMEWW method comparison, separate samples were spiked at a low level inoculum targeting 25 CFU/100mL and a high level

inoculum targeting 75 CFU/100mL. For the ISO method comparison, separate samples were spiked at a low level inoculum

targeting 25 CFU/250mL and a high level inoculum targeting 75 CFU/250mL. After a two-week equilibration, waters were

analyzed by the methods described below. For both comparisons, the 3M Petrifilm EC Plates were pre-hydrated with 1mL of

sterile deionized water, held at refrigerated temperature for no longer than a week, and allowed to come to room temperature

prior to analysis. In order to achieve a countable range, ten-fold serial dilutions were made using filter sterilized (22μm) dilution

blanks of each brand of water.


For the SMEWW method comparison, 100mL portions of inoculated samples were filtered in duplicate through MCE

membrane filters and placed onto 3M Petrfilm EC Plates and endo LES agar plates. 3M Petrfilm EC Plates were incubated

for 24 ± 2 hours at 35 ± 0.5°C and endo LES agar plates were incubated for 22–24 hours at 35 ± 0.5°C. After incubation, 3M

Petrfilm EC Plates were examined for typical colonies (blue or red, producing gas bubbles) and atypical colonies (blue or red,

without gas production), and endo LES agar plates were observed for colonies that were pink to dark red with a green metallic

surface sheen. One typical colony (for endo LES agar and 3M Petrfilm EC Plates) and two atypical colonies (3M Petrfilm

EC Plates only) per sample were picked to Lauryl Tryptose Broth (LTB) tubes and incubated at 35 ± 1°C for 48 ± 2 hours.

After incubation, LTB tubes were examined for gas production, and a loopful of broth from positive tubes were transferred

to Brilliant Green Lactose Bile (BGLB) tubes (for the analysis of the coliform bacteria) and incubated at 35 ± 1°C for 48 ±

2 hours and to EC broth (for the analysis of E. coli) and incubated at 45.5 ± 0.2°C for 48 ± 2 hours. After incubation, tubes

were observed for gas production. Samples producing gas in BGLB tubes were recorded as positive. Samples producing

gas in EC broth tubes were streaked to Levine’s Eosin-Methylene Blue agar (l-EMB) and incubated for 18–24 hours at 35 ±

1°C. Plates were examined for typical colonies (red or pink with green metallic sheen). Typical colonies were transferred to

Tryptone broth, MR-VP broth, Kosers’ Citrate broth, and LTB and incubated at 35 ± 1°C. After 24 hours, a drop of Kovacs’

reagent was added to the Tryptone broth to test for the presence of indole. Appearance of a distinct red color in the upper layer

was recorded as positive. After 48 ± 2 hours of incubation, a 1mL aliquot was removed from the MR-VP tubes and the tubes

were reincubated for an additional 48 ± 2 hours. 0.6mL of α-naphthol, 0.2mL of 40% KOH, and a few crystals of creatine

were added to the 1mL aliquot, allowed to incubate at room temperature for 2 hours. A positive reaction was recorded if eosin

pink color developed. After the additional 48 hour incubation, five drops of methyl red solution were added to each MR-VP

tube and a distinct color change to red was indicative of a positive reaction. After a 96 ± 4 hour incubation the Kosers’ Citrate

tubes were removed and observed for turbidity. An abscess of turbidity indicated a negative reaction. Colonies that produced

reactions typical of E. coli and coliforms were recorded as such and statistical analysis was conducted.

B. ISO Method comparison.

250mL portions of inoculated samples were filtered in duplicate through Mixed Cellulose Ester (MCE) membrane filters

and placed onto 3M Petrfilm EC Plates or through Cellulose Nitrate (CN) membrane filters and placed onto Lactose TTC

agar with Tergitol 7 plates. 3M Petrfilm EC Plates were incubated at 36 ± 2°C for 24 ± 2 hours and Lactose TTC plates were

incubated at 36 ± 2°C for 21 ± 3 hours. After incubation 3M Petrfilm EC Plates were examined for typical colonies (blue or

red, producing gas bubbles) and atypical colonies (blue or red, without gas production) and Lactose TTC plates were observed

for yellow to yellow-orange colonies with yellow acid zones. One typical colony (Lactose TTC and 3M Petrfilm EC Plate) and

two atypical colonies (3M Petrfilm EC Plate only) per sample were picked to TSA and incubated at 36 ± 2°C for 24 ± 2 hours.

Representative colonies on the TSA were confirmed according to the ISO reference method by conducting the spot Oxidase

test. Additionally, a typical colony was picked and inoculated into 3 mL tryptophan broth. Broth was incubated at 44 ± 0.5°C

for 21 ± 3 hours, then 0.2–0.3mL of Kovak’s reagent was added to test for the presence of indole. Samples that developed

a cherry-red color at the surface of the broth were recorded as positive for indole production. Those colonies that produced

oxidase negative and indole negative results were recorded as coliforms. Colonies that produced oxidase negative and indole

positive results were recorded as E. coli.

All confirmed (typical and confirmed atypical) counts were used in the data analysis. Detailed Results are presented in Table 5

of Appendix 1.

Yeast and Mold Plate Count

Two lots of all four types of water samples were inoculated with one of three organisms (Penicillium sp. ATCC #18307,

Paecilomyces sp. ATCC #1114, and Candida albicans ATCC #10231) as specified in Table 1 of Appendix 1. Separate samples

were spiked at a low level inoculum targeting 25 CFU/100mL and a high level inoculum targeting 75 CFU/100mL and were

allowed to equilibrate overnight at room temperature. In order to achieve a countable range, ten-fold serial dilutions were made

using filter sterilized (22μm) dilution blanks of each brand of water. Samples were analyzed by membrane filtration for the

analysis of yeast and molds by both the SMEWW method and a customer method using acidified Potato Dextrose agar (aPDA).


100mL portions of inoculated samples were filtered in duplicate through MCE membrane filters and placed onto 3M Petrifilm

Aqua AQYM Plates and Cooke Rose Bengal agar (CRB) plates. After filtration, membrane filters were placed onto the 3M

Petrifilm Aqua AQYM Plates and 1mL of sterile deionized water was placed on top of the filter and the hydration fluid was

spread on the plate. 3M Petrifilm Aqua AQYM Plates and CRB plates were incubated for 5 days at 20 ± 1°C. Plates with

yeast and mold colonies in the countable range were read and the data recorded at both 3 and 5 days.

B. Customer Method comparison.

100mL portions of inoculated samples were filtered in duplicate through MCE membrane filters and placed onto 3M Petrifilm

Aqua AQYM Plates and aPDA plates. After filtration, membrane filters were placed onto the 3M Petrifilm Aqua AQYM Plates,

1mL of sterile deionized water was placed on top of the filter, and the hydration fluid was spread on the plate. 3M Petrifilm

Aqua AQYM Plates and aPDA plates were incubated at 25 ± 1°C for 5 days. Plates with yeast and mold colonies in the

countable range were read and the data recorded at both 3 and 5 days. Detailed results are presented in Table 6 of Appendix 1.

Statistical Analysis

Plates with 1–250 colonies were considered for use in the analysis on the heterotrophic plates, 1–150 on the coliform plates/agars,

1–100 on the EB plates/agars, and 1–150 for the yeast and mold plates/agars. If none of the plates had the minimum number of

colonies, the exact count on the least dilute test was selected for analysis. Procedures described by the various standards were

used to calculate colony counts. Specifically, a weighted average was reported for each heterotrophic test portion when the ISO

method was used as the reference. These counts were converted to log10

counts to more nearly match the underlying assumption of

a normal distribution. Indefinite values (<1 colony forming unit/sample) were treated as missing values in the analysis. Analysis of

variance was used to calculate the method repeatability. Repeatability variances were then compared using an F-ratio test. A paired

t test (by inoculation level) was used to compare the differences in counts between the Petrifilm Plate methods and the reference

methods. In all statistical tests, a resulting value of p<0.05 was taken to indicate a significant difference.

Results

Two separate lots of four types of bottled water (Purified, Regional Spring, Natural Spring, and Carbonated Natural Spring) were

evaluated for heterotrophic bacteria, coliforms, Enterobacteriaceae and yeast and mold by the 3M Petrifilm Aqua Plate methods

and the corresponding reference method.

Heterotrophic plate method

Heterotrophic counts were obtained using the 3M Petrifilm Aqua AQHC Plate method and the PCA method (SMEWW). At

the uninoculated level, the mean log heterotrophic count from the 3M Petrifilm Aqua AQHC Plate method was not statistically

different from the mean log count from the PCA method (p-value = 0.488). At the low and high inoculation levels, the two

methods were also similar (p-values equal to 0.985 and 0.917, respectively) (see Table 7).

Heterotrophic counts were also obtained using the YEA method (ISO). At the low inoculation level, the mean log heterotrophic

count per 1mL direct on the 3M Petrifilm Aqua AQHC Plate was not significantly different from the mean log heterotrophic

count using the YEA method (p-value = 0.637). At the high inoculation level, there was no statistical difference between the two

methods (p-value = 0.204) (see Table 7).

When comparing the 3M Petrifilm Aqua AQHC Plate method with an incubation temperature of 22°C and the YEA method

with an incubation temperature of 22°C, there was no statistical difference at any of the inoculation levels – uninoculated, low,

and high (p-values equal to 0.463, 0.637, and 0.202, respectively) (see Table 7).

Heterotrophic counts were obtained using the 3M Petrifilm Aqua AQHC Plate method and the PCA method (SMEWW) with

membrane filtration. The mean log heterotrophic counts per filter were not significantly different between the 3M Petrifilm Aqua

AQHC Plate method and the PCA method at either the uninoculated level (p-value = 0.212), the low inoculation level (p-value =

0.833), or at the high inoculation level (p-value = 0.603) (see Table 7).

The repeatability variances of the 3M Petrifilm Aqua AQHC Plate method were not significantly different from that of the

reference methods, except in the case of the comparison between the 3M Petrifilm Aqua AQHC Plate method and the PCA

method with direct plating and an incubation period of 36°C. In that comparison the repeatability variance of the 3M Petrifilm

Aqua AQHC Plate method was better than that of the PCA method (see Table 7).

Coliform plate method

Coliform counts were obtained using the 3M Petrifilm Aqua AQCC Plate method and the endo LES agar method (SMEWW).

The mean log coliform count per filter was not significantly different between the 3M Petrifilm Aqua AQCC Plate method and

the endo LES agar method at the low inoculation level (p-value = 0.403). At the high inoculation level, the mean log coliform

count per filter was significantly different between that obtained from the 3M Petrifilm Aqua AQCC Plate method and the mean

log count from the endo LES agar method (p = 0.016). On average, the log counts from the 3M Petrifilm Aqua AQCC Plate

were greater in value than those for the endo LES agar method (see Table 7).

The repeatability variance of the 3M Petrifilm Aqua AQCC Plate method was significantly better from that of the endo LES agar

method for the low inoculation level. For the high inoculation level, the repeatability variance of the 3M Petrifilm Aqua AQCC

Plate method was not significantly different from that of the endo LES agar method (see Table 7).

Coliform counts were obtained using the 3M Petrifilm Aqua AQCC Plate method and the Lactose TTC with Tergitol 7 agar

method (ISO). The mean log coliform counts per filter were not significantly different between the 3M Petrifilm Aqua AQCC

Plate method and the Lactose TTC with Tergitol 7 agar method at the low inoculation level (p-value = 0.495) and the high

inoculation level (p-value = 0.638) (see Table 7).

As was the case with the comparison between the 3M Petrifilm Aqua AQCC Plate method and the endo LES agar method, the

repeatability variances were better and not different (low level and high level, respectively) between the 3M Petrifilm Aqua

AQCC Plate method and the Lactose TTC with Tergitol 7 agar method (see Table 7).

Enterobacteriaceae plate method

Enterobacteriaceae counts were obtained using the 3M Petrifilm Aqua AQEB Plate method and VRBG agar method, a customer

method. The mean log Enterobacteriaceae counts per filter were not significantly different between the 3M Petrifilm Aqua

AQEB Plate method and the VRBG agar method at the low inoculation level (p-value = 0.844) and the high inoculation level

(p-value = 0.47) (see Table 7).

The repeatability variances of the 3M Petrifilm Aqua AQEB Plate method were not significantly different from those of the

VRBG method for the high level of inoculation. The repeatability variance of the VRBG agar method was better than that of the

3M Petrifilm Aqua AQEB Plate method for the low inoculation level (see Table 7).

Yeast and Mold plate method

Yeast and Mold counts were obtained using the 3M Petrifilm Aqua AQYM Plate method and Cooke Rose Bengal agar (CRB)

– SMEWW reference agar method – and acidified Potato Dextrose agar (aPDA), a customer method. The mean log yeast and

mold counts per filter were not significantly different between the 3M Petrifilm Aqua AQYM Plate method and the CRB agar

method at the low inoculation level at day 3 (p-value = 0.745) and the high inoculation level at day 3 (p-value = 0.063). The

mean log yeast and mold counts per filter were not significantly different between the 3M Petrifilm Aqua AQYM Plate method

and the CRB agar method at the low inoculation level at day 5 (p-value = 0.121) and the high inoculation level at day 5 (p-value

= 0.052) (see Table 7).

The mean log yeast and mold counts per filter were not significantly different between the 3M Petrifilm Aqua AQYM Plate

method and the aPDA agar method at the low inoculation level at day 3 (p-value = 0.844) and the high inoculation level at day 3

(p-value = 0.723). The mean log yeast and mold counts per filter were not significantly different between the 3M Petrifilm Aqua

AQYM Plate method and the aPDA agar method at the low inoculation level at day 5 (p-value = 0.506). At the high inoculation

level at day 5, the mean log yeast and mold counts per filter were significantly different between the 3M Petrifilm Aqua AQYM

Plate method and the aPDA method (p = 0.036) with the log counts per filter for the 3M Petrifilm Aqua AQYM Plate method, on

average, greater in value than those from the aPDA method (see Table 7).

The repeatability variances of the 3M Petrifilm Aqua AQYM Plate method were not significantly different from those of the

reference methods for the low and high levels of inoculation when the plates were enumerated at three days. When the plates

were enumerated after five days, the repeatability variances were also not different between the 3M Petrifilm Aqua AQYM Plate

method and the reference methods at the high level of inoculation. The repeatability variances of the 3M Petrifilm Aqua AQYM

Plate method were significantly better than those from the reference methods at the low level of inoculation when the plates were

enumerated after five days (see Table 7).

Discussion

The results of this independent evaluation demonstrate the reliability of the 3M Petrifilm Plate methods as an easy-to-use

alternative to the reference methods for the analysis of bacteria, yeast and molds in bottled water by direct plate method or

membrane filtration. All five types of 3M Petrifilm Plates reduce the cost of sample preparation by eliminating the need for

producing and sterilizing reference method agars. 3M Petrifilm Aqua AQHC Plates, 3M Petrifilm Aqua AQCC Plates, and 3M

Petrifilm Aqua AQEB Plates increase sample productivity with the ability to rehydrate the plates and store them at refrigeration

temperature for up to a week. Additionally, the 3M Petrifilm Aqua AQCC Plate method may eliminate the need for lengthy

confirmation of coliforms by producing colored colonies with gas production compared to the reference agars. Similarly, the

3M Petrifilm E. coli/Coliform Plate method may eliminate the need for lengthy confirmation of coliforms and of E. coli by

producing distinctly colored colonies with gas production compared to the reference agars.

3M and Petrifilm are trademarks of 3M.

APPENDIX 1

Tabl

e 1:

Org

anis

m a

nd W

ater

Inoc

ulat

ion

Guid

elin

e

Wat

er T

ype

Sam

ple

Hete

rotr

ophi

c Pl

ate

(AQH

C)Co

lifor

m P

late

(AQC

C)Es

cher

ichi

a co

li Pl

ate

(EC)

Ente

roba

cter

iace

ae P

late

(AQE

B)Ye

ast a

nd M

old

Plat

e (A

QYM

)

Purifi

ed

Dasa

ni, L

ot 1

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Pe

nici

llium

sp.

ATCC

183

07

Dasa

ni, L

ot 2

Ente

roco

ccus

faec

ium

ATCC

194

34Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22En

tero

bact

er a

erog

enes

ATCC

#35

029

Paec

ilom

yces

sp.

ATCC

111

4

Nest

le P

ure

Life

, Lot

1Ps

eudo

mon

as a

erug

inos

aAT

CC 1

5442

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Paec

ilom

yces

sp.

ATCC

111

4

Nest

le P

ure

Life

, Lot

2Es

cher

ichi

a co

liAT

CC 2

5922

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Esch

eric

hia

coli

ATCC

259

22Pe

nici

llium

sp.

ATCC

183

07

Aqua

fina,

Lot

1Es

cher

ichi

a co

liAT

CC 2

5922

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Pa

ecilo

myc

es s

p.AT

CC 1

114

Aqua

fina,

Lot

2En

tero

cocc

us fa

eciu

mAT

CC 1

9434

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Cand

ida

albi

cans

ATCC

102

31

Regi

onal

Spr

ing

#1Ic

e M

ount

ain,

Lot

1Es

cher

ichi

a co

liAT

CC 2

5922

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9En

tero

bact

er a

erog

enes

ATCC

#35

029

Esch

eric

hia

coli

ATCC

259

22Ca

ndid

a al

bica

nsAT

CC 1

0231

Ice

Mou

ntai

n, L

ot 2

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2eum

onia

ePe

nici

llium

sp.

ATCC

183

07

Regi

onal

Spr

ing

#2Kr

oger

Tre

ated

, Lot

1Es

cher

ichi

a co

liAT

CC 2

5922

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Esch

eric

hia

coli

ATCC

259

22Pa

ecilo

myc

es s

p.AT

CC 1

114

Krog

er T

reat

ed, L

ot 2

Ente

roco

ccus

faec

ium

ATCC

194

34En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Ca

ndid

a al

bica

nsAT

CC 1

0231

Regi

onal

Spr

ing

#3Tr

auth

, Lot

1Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Pe

nici

llium

sp.

ATCC

183

07

Trau

th, L

ot 2

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Es

cher

ichi

a co

liAT

CC 2

5922

Paec

ilom

yces

sp.

ATCC

111

4

Natu

ral S

prin

g

Evia

n, L

ot 1

Ente

roco

ccus

faec

ium

ATCC

194

34En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Pe

nici

llium

sp.

ATCC

183

07

Evia

n, L

ot 2

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22En

tero

bact

er a

erog

enes

ATCC

#35

029

Cand

ida

albi

cans

ATCC

102

31

Jana

, Lot

1Es

cher

ichi

a co

liAT

CC 2

5922

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9En

tero

bact

er a

erog

enes

ATCC

#35

029

Esch

eric

hia

coli

ATCC

259

22Pa

ecilo

myc

es s

p.AT

CC 1

114

Jana

, Lot

2Ps

eudo

mon

as a

erug

inos

aAT

CC 1

5442

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Peni

cilliu

m s

p.AT

CC 1

8307

Fiji,

Lot

1En

tero

cocc

us fa

eciu

mAT

CC 1

9434

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22Pa

ecilo

myc

es s

p.AT

CC 1

114

Fiji,

Lot

2Es

cher

ichi

a co

liAT

CC 2

5922

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Pe

nici

llium

sp.

ATCC

183

07

Voss

, Lot

1En

tero

cocc

us fa

eciu

mAT

CC 1

9434

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22Ca

ndid

a al

bica

nsAT

CC 1

0231

Voss

, Lot

2Ps

eudo

mon

as a

erug

inos

aAT

CC 1

5442

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Pe

nici

llium

sp.

ATCC

183

07

Ty N

ant S

prin

g, L

ot 1

Ente

roco

ccus

faec

ium

ATCC

194

34Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2En

tero

bact

er a

erog

enes

ATCC

#35

029

Cand

ida

albi

cans

ATCC

102

31

Ty N

ant S

prin

g, L

ot 2

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Es

cher

ichi

a co

liAT

CC 2

5922

Peni

cilliu

m s

p.AT

CC 1

8307

Tabl

e 1

(con

tinue

d):

Orga

nism

and

Wat

er In

ocul

atio

n Gu

idel

ine

Wat

er T

ype

Sam

ple

Hete

rotr

ophi

c Pl

ate

(AQH

C)Co

lifor

m P

late

(AQC

C)Es

cher

ichi

a co

li Pl

ate

(EC)

Ente

roba

cter

iace

ae P

late

(AQE

B)Ye

ast a

nd M

old

Plat

e (A

QYM

)

Natu

ral S

prin

g,ca

rbon

ated

Gero

lste

iner

, Lot

1Es

cher

ichi

a co

liAT

CC 2

5922

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9En

tero

bact

er a

erog

enes

ATCC

#35

029

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Peni

cilliu

m s

p.AT

CC 1

8307

Gero

lste

iner

, Lot

2En

tero

cocc

us fa

eciu

mAT

CC 1

9434

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Kleb

siel

la p

neum

onia

ATCC

# 4

352

Esch

eric

hia

coli

ATCC

259

22Ca

ndid

a al

bica

nsAT

CC 1

0231

Apol

linar

is, L

ot 1

Ente

roco

ccus

faec

ium

ATCC

194

34En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Pa

ecilo

myc

es s

p.AT

CC 1

114

Apol

linar

is, L

ot 2

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2En

tero

bact

er a

erog

enes

ATCC

#35

029

Cand

ida

albi

cans

ATCC

102

31

Perri

er, L

ot 1

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Ca

ndid

a al

bica

nsAT

CC 1

0231

Perri

er, L

ot 2

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42En

tero

bact

er a

erog

enes

ATCC

#35

029

Ente

roba

cter

aer

ogen

esAT

CC #

3502

9En

tero

bact

er a

erog

enes

ATCC

#35

029

Paec

ilom

yces

sp.

ATCC

111

4

Voss

Spa

rklin

g, L

ot 1

Ente

roco

ccus

faec

ium

ATCC

194

34Es

cher

ichi

a co

liAT

CC 2

5922

Esch

eric

hia

coli

ATCC

259

22Es

cher

ichi

a co

liAT

CC 2

5922

Paec

ilom

yces

sp.

ATCC

111

4

Voss

Spa

rklin

g, L

ot 2

Pseu

dom

onas

aer

ugin

osa

ATCC

154

42Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2Kl

ebsi

ella

pne

umon

iaAT

CC #

435

2En

tero

bact

er a

erog

enes

ATCC

#35

029

Cand

ida

albi

cans

ATCC

102

31

Tabl

e 2:

Res

ults

for A

QHC

vs. S

MEW

W (P

CA) a

nd IS

O (Y

EA) b

y Di

rect

Pla

ting

(CFU

/mL)

and

Mem

bran

e Fi

ltrat

ion

(CFU

/100

mL)

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Dire

ct P

latin

gM

embr

ane

Filtr

atio

n

Avg.

AQ

HC

(36° C

)

Log

Avg.

AQ

HC

(36° C

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

Avg.

YEA

(3

6° C)

Log

Avg.

YE

A (3

6° C)

Avg.

AQ

HC

(22° C

)

Log

Avg.

AQ

HC

(22° C

)Av

g. Y

EA

(22° C

)

Log

Avg.

YE

A (2

2° C)

Avg.

AQH

C (3

6° C)

(SM

EWW

)

Log

Avg.

AQH

C (3

6° C)

(SM

EWW

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

(SM

EWW

)

P. ae

rugi

nosa

1Da

sani

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Da

sani

Low

171.

2297

231.

3613

211.

3202

211.

3217

221.

3314

281.

4460

271.

4169

1Da

sani

High

521.

7147

601.

7776

581.

7593

571.

7519

591.

7708

791.

8975

881.

9404

E. fa

eciu

m

2Da

sani

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Da

sani

Low

501.

6919

481.

6809

561.

7471

541.

7282

531.

7192

521.

7093

601.

7720

2Da

sani

High

122

2.08

4312

72.

1033

114

2.05

6511

52.

0586

110

2.03

9311

22.

0491

119

2.07

20

P. ae

rugi

nosa

1Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Ne

stle

Pur

elife

Low

211.

3217

231.

3469

271.

4311

261.

4137

271.

4302

261.

4098

221.

3295

1Ne

stle

Pur

elife

High

731.

8613

631.

7955

651.

8116

641.

8028

641.

8049

731.

8627

751.

8741

E. c

oli

2Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Ne

stle

Pur

elife

Low

401.

5957

471.

6674

461.

6559

441.

6424

461.

6618

451.

6470

461.

6611

2Ne

stle

Pur

elife

High

881.

9427

921.

9634

901.

9517

841.

9216

881.

9440

861.

9344

861.

9334

E. c

oli

1Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

1Aq

uafin

aLo

w53

1.72

2342

1.62

2844

1.64

2449

1.68

8745

1.64

9344

1.63

6241

1.60

74

1Aq

uafin

aHi

gh83

1.91

5594

1.97

0886

1.92

7885

1.92

9191

1.95

5882

1.91

0574

1.86

86

Tabl

e 2

(con

tinue

d):

Resu

lts fo

r AQH

C vs

. SM

EWW

(PCA

) and

ISO

(YEA

) by

Dire

ct P

latin

g (C

FU/m

L) a

nd M

embr

ane

Filtr

atio

n (C

FU/1

00m

L)

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Dire

ct P

latin

gM

embr

ane

Filtr

atio

n

Avg.

AQ

HC

(36° C

)

Log

Avg.

AQ

HC

(36° C

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

Avg.

YEA

(3

6° C)

Log

Avg.

YE

A (3

6° C)

Avg.

AQ

HC

(22° C

)

Log

Avg.

AQ

HC

(22° C

)Av

g. Y

EA

(22° C

)

Log

Avg.

YE

A (2

2° C)

Avg.

AQH

C (3

6° C)

(SM

EWW

)

Log

Avg.

AQH

C (3

6° C)

(SM

EWW

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

(SM

EWW

)

E. fa

eciu

m

2Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

2Aq

uafin

aLo

w59

1.76

9952

1.71

1364

1.79

2749

1.68

5555

1.73

8659

1.76

9954

1.73

05

2Aq

uafin

aHi

gh11

42.

0545

121

2.07

8611

92.

0755

116

2.06

1910

72.

0293

128

2.10

6713

42.

1254

E. c

oli

1Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Ic

e M

ount

ain

Low

461.

6623

401.

5986

391.

5854

461.

6567

461.

6602

521.

7093

451.

6528

1Ic

e M

ount

ain

High

101

2.00

4195

1.97

5194

1.97

0890

1.95

4099

1.99

4297

1.98

4491

1.95

61

P. ae

rugi

nosa

2Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Ic

e M

ount

ain

Low

251.

3779

301.

4667

291.

4614

251.

3869

251.

3891

251.

3869

321.

4978

2Ic

e M

ount

ain

High

551.

7342

601.

7719

561.

7434

581.

7628

551.

7349

741.

8660

811.

9054

E. c

oli

1Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Kr

oger

Tre

ated

Low

441.

6417

491.

6898

461.

6535

551.

7401

591.

7671

451.

6515

511.

7022

1Kr

oger

Tre

ated

High

831.

9155

851.

9262

831.

9190

931.

9684

871.

9395

831.

9186

771.

8816

P. ae

rugi

nosa

2Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Kr

oger

Tre

ated

Low

631.

7988

531.

7223

551.

7359

641.

8043

601.

7766

531.

7197

561.

7471

2Kr

oger

Tre

ated

High

103

2.00

9611

12.

0453

114

2.05

6810

42.

0169

111

2.04

1810

42.

0165

108

2.03

31

E. c

oli

1Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Tr

auth

Tre

ated

Low

471.

6696

451.

6470

451.

6461

521.

7140

441.

6430

451.

6481

471.

6712

1Tr

auth

Tre

ated

High

104

2.01

6993

1.96

8498

1.98

9095

1.97

5310

22.

0062

921.

9637

881.

9422

P. ae

rugi

nosa

2Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Tr

auth

Tre

ated

Low

251.

3948

221.

3295

251.

3891

251.

3779

211.

3106

351.

5396

311.

4828

2Tr

auth

Tre

ated

High

731.

8613

651.

8080

611.

7800

631.

7955

571.

7504

831.

9188

931.

9685

Natu

ral

1Ev

ian

10^

04

0.53

964

0.58

804

0.53

963

0.34

954

0.60

2132

1.49

3839

1.58

51

1Ev

ian

10^

-10

0.00

000

0.00

000

0.00

000

0.00

000

0.00

004

0.53

965

0.65

05

1Ev

ian

10^

-20

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

Natu

ral

2Ev

ian

10^

02

0.30

102

0.30

102

0.23

863

0.38

911

0.00

0013

1.09

6610

0.97

71

2Ev

ian

10^

-10

0.00

000

0.00

000

0.00

000

0.00

000

0.00

001

0.00

002

0.15

05

2Ev

ian

10^

-20

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

E. fa

eciu

m

1Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Ev

ian

Low

581.

7593

631.

7988

581.

7549

551.

7397

571.

7491

421.

6173

461.

6611

1Ev

ian

High

106

2.02

4811

82.

0716

116

2.06

2411

32.

0528

113

2.05

1711

72.

0658

111

2.04

33

E. c

oli

2Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Ev

ian

Low

441.

6424

461.

6623

451.

6505

491.

6839

411.

6123

471.

6720

541.

7321

2Ev

ian

High

891.

9469

871.

9393

911.

9581

921.

9609

871.

9394

871.

9370

911.

9566

Natu

ral

1Ja

na10

^0

141.

1276

151.

1722

151.

1761

100.

9978

131.

1127

113

2.05

0410

22.

0076

1Ja

na10

^-1

10.

0000

20.

1505

10.

0000

20.

1505

10.

0000

121.

0570

111.

0396

1Ja

na10

^-2

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

10.

0000

20.

1505

Tabl

e 2

(con

tinue

d):

Resu

lts fo

r AQH

C vs

. SM

EWW

(PCA

) and

ISO

(YEA

) by

Dire

ct P

latin

g (C

FU/m

L) a

nd M

embr

ane

Filtr

atio

n (C

FU/1

00m

L)

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Dire

ct P

latin

gM

embr

ane

Filtr

atio

n

Avg.

AQ

HC

(36° C

)

Log

Avg.

AQ

HC

(36° C

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

Avg.

YEA

(3

6° C)

Log

Avg.

YE

A (3

6° C)

Avg.

AQ

HC

(22° C

)

Log

Avg.

AQ

HC

(22° C

)Av

g. Y

EA

(22° C

)

Log

Avg.

YE

A (2

2° C)

Avg.

AQH

C (3

6° C)

(SM

EWW

)

Log

Avg.

AQH

C (3

6° C)

(SM

EWW

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

(SM

EWW

)

Natu

ral

2Ja

na10

^0

139

2.14

0812

32.

0880

136

2.13

1912

52.

0949

130

2.11

08TN

TC*

NATN

TCNA

2Ja

na10

^-1

131.

0880

131.

0938

131.

0792

121.

0777

141.

1276

129

2.10

9914

32.

1552

2Ja

na10

^-2

10.

0000

10.

0000

20.

1505

20.

1505

10.

0000

131.

1127

131.

0880

E. c

oli

1Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

1Ja

naLo

w41

1.60

2150

1.68

8144

1.63

3645

1.65

2245

1.64

5044

1.63

8242

1.61

73

1Ja

naHi

gh86

1.93

4276

1.88

0885

1.92

8388

1.93

9984

1.92

3884

1.92

3581

1.90

54

P. ae

rugi

nosa

2Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

2Ja

naLo

w21

1.32

0225

1.38

6926

1.40

5822

1.34

0623

1.34

6935

1.53

9641

1.60

95

2Ja

naHi

gh62

1.79

1557

1.75

5661

1.78

1061

1.77

6355

1.73

5582

1.91

0574

1.86

82

Natu

ral

1Fi

ji10

^0

90.

9225

60.

7526

90.

9287

80.

8891

70.

8010

731.

8613

681.

8287

1Fi

ji10

^-1

10.

0000

00.

0000

10.

0000

00.

0000

00.

0000

80.

8741

80.

8495

1Fi

ji10

^-2

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

Natu

ral

2Fi

ji10

^0

200

2.29

9821

32.

3284

209

2.32

0119

62.

2919

189

2.27

59TN

TCNA

TNTC

NA

2Fi

ji10

^-1

231.

3512

231.

3469

191.

2788

181.

2546

211.

3106

TNTC

NATN

TCNA

2Fi

ji10

^-2

30.

3891

20.

3010

40.

3891

20.

3010

30.

3891

231.

3613

231.

3469

E. fa

eciu

m

1Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

1Fi

jiLo

w53

1.72

4049

1.69

0255

1.73

4256

1.74

2162

1.79

1557

1.75

5653

1.71

86

1Fi

jiHi

gh11

42.

0545

112

2.04

8110

92.

0374

114

2.05

3511

42.

0568

115

2.05

9910

22.

0063

E. c

oli

2Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

2Fi

jiLo

w44

1.64

0637

1.56

1945

1.64

7046

1.66

2747

1.66

6252

1.71

5343

1.62

69

2Fi

jiHi

gh96

1.98

1794

1.97

0587

1.93

9585

1.92

6894

1.97

2977

1.88

3486

1.93

18

Natu

ral

1Vo

ss10

^0

531.

7230

611.

7848

511.

7022

581.

7593

551.

7404

TNTC

NATN

TCNA

1Vo

ss10

^-1

60.

7386

80.

8741

50.

6276

60.

7236

70.

8010

571.

7553

471.

6657

1Vo

ss10

^-2

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

70.

8010

60.

7236

Natu

ral

2Vo

ss10

^0

141.

1276

131.

1021

131.

0966

121.

0731

121.

0792

192

2.28

0916

62.

2188

2Vo

ss10

^-1

20.

1505

10.

0000

00.

0000

10.

0000

10.

0000

161.

1963

131.

1087

2Vo

ss10

^-2

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

20.

3010

20.

1505

E. fa

eciu

m

1Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

1Vo

ssLo

w63

1.79

8861

1.78

4856

1.74

3456

1.74

2959

1.76

9353

1.71

6852

1.71

08

1Vo

ssHi

gh8

0.87

4112

1.06

038

0.88

9111

52.

0601

107

2.02

7311

32.

0504

124

2.09

16

P. ae

rugi

nosa

2Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

2Vo

ssLo

w27

1.42

8725

1.38

6923

1.34

6922

1.33

8323

1.35

2122

1.32

9528

1.44

69

2Vo

ssHi

gh68

1.82

9261

1.78

1458

1.75

6955

1.73

5555

1.73

5982

1.91

3592

1.96

37

Natu

ral

1Vo

ss S

park

ling

10^

00

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

**-

NT-

1Vo

ss S

park

ling

10^

-10

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

1Vo

ss S

park

ling

10^

-20

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

Tabl

e 2

(con

tinue

d):

Resu

lts fo

r AQH

C vs

. SM

EWW

(PCA

) and

ISO

(YEA

) by

Dire

ct P

latin

g (C

FU/m

L) a

nd M

embr

ane

Filtr

atio

n (C

FU/1

00m

L)

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Dire

ct P

latin

gM

embr

ane

Filtr

atio

n

Avg.

AQ

HC

(36° C

)

Log

Avg.

AQ

HC

(36° C

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

Avg.

YEA

(3

6° C)

Log

Avg.

YE

A (3

6° C)

Avg.

AQ

HC

(22° C

)

Log

Avg.

AQ

HC

(22° C

)Av

g. Y

EA

(22° C

)

Log

Avg.

YE

A (2

2° C)

Avg.

AQH

C (3

6° C)

(SM

EWW

)

Log

Avg.

AQH

C (3

6° C)

(SM

EWW

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

(SM

EWW

)

Natu

ral

2Vo

ss S

park

ling

10^

00

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

2Vo

ss S

park

ling

10^

-10

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

2Vo

ss S

park

ling

10^

-20

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

E. fa

eciu

m

1Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Vo

ss S

park

ling

Low

471.

6668

451.

6450

541.

7324

541.

7312

491.

6902

591.

7699

511.

7031

1Vo

ss S

park

ling

High

126

2.09

8511

52.

0604

117

2.06

6611

52.

0586

111

2.04

4912

82.

1051

134

2.12

70

P. ae

rugi

nosa

2Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Vo

ss S

park

ling

Low

251.

3869

291.

4621

311.

4842

281.

4447

271.

4232

471.

6712

431.

6316

2Vo

ss S

park

ling

High

591.

7664

651.

8116

641.

8049

581.

7593

551.

7359

991.

9933

104

2.01

69

Natu

ral

1Ty

Nan

t10

^0

164

2.21

4315

22.

1817

172

2.23

3116

62.

2186

157

2.19

50TN

TCNA

TNTC

NA

1Ty

Nan

t10

^-1

201.

2899

151.

1548

171.

2304

191.

2733

181.

2546

TNTC

NATN

TCNA

1Ty

Nan

t10

^-2

30.

3891

20.

1505

20.

3010

20.

3010

30.

3891

191.

2657

171.

2274

Natu

ral

2Ty

Nan

t10

^0

371.

5668

321.

5051

411.

6095

411.

6127

441.

6378

TNTC

NATN

TCNA

2Ty

Nan

t10

^-1

50.

6505

50.

6276

60.

7236

50.

6505

40.

6021

281.

4387

261.

4098

2Ty

Nan

t10

^-2

00.

0000

10.

0000

00.

0000

00.

0000

00.

0000

40.

5396

40.

5396

E. fa

eciu

m

1Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

1Ty

Nan

tLo

w63

1.79

2754

1.72

9750

1.69

8654

1.72

6060

1.77

4557

1.75

5956

1.74

34

1Ty

Nan

tHi

gh12

22.

0843

107

2.02

7210

62.

0234

113

2.05

2510

72.

0263

991.

9933

108

2.03

33

P. ae

rugi

nosa

2Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

2Ty

Nan

tLo

w22

1.32

9526

1.40

9831

1.48

4229

1.45

4231

1.48

4245

1.65

1549

1.68

55

2Ty

Nan

tHi

gh61

1.78

0062

1.78

8555

1.73

9259

1.76

9955

1.73

5970

1.84

4466

1.81

41

Natu

ral

1Ge

rols

tein

er10

^0

00.

000

00.

000

00.

000

00.

000

00.

000

NT-

NT-

1Ge

rols

tein

er10

^-1

00.

000

00.

000

00.

000

00.

000

00.

000

NT-

NT-

1Ge

rols

tein

er10

^-2

00.

000

00.

000

00.

000

00.

000

00.

000

NT-

NT-

Natu

ral

2Ge

rols

tein

er10

^0

00.

000

00.

000

00.

000

00.

000

00.

000

NT-

NT-

2Ge

rols

tein

er10

^-1

00.

000

00.

000

00.

000

00.

000

00.

000

NT-

NT-

2Ge

rols

tein

er10

^-2

00.

000

00.

000

00.

000

00.

000

00.

000

NT-

NT-

E. c

oli

1Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

1Ge

rols

tein

erLo

w44

1.64

1741

1.64

1744

1.60

7442

1.63

8248

1.61

5544

1.64

0652

1.71

08

1Ge

rols

tein

erHi

gh90

1.95

4078

1.95

4083

1.89

2194

1.91

8882

1.97

2910

01.

9997

961.

9797

E. fa

eciu

m

2Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

000

0.00

00

2Ge

rols

tein

erLo

w61

1.78

0055

1.78

0055

1.73

5556

1.73

9253

1.74

2146

1.66

1152

1.71

08

2Ge

rols

tein

erHi

gh11

02.

0394

101

2.03

9410

82.

0041

104

2.03

1410

42.

0145

126

2.10

0411

02.

0394

Natu

ral

1Ap

ollin

aris

10^

00

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

1Ap

ollin

aris

10^

-10

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

1Ap

ollin

aris

10^

-20

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

Tabl

e 2

(con

tinue

d):

Resu

lts fo

r AQH

C vs

. SM

EWW

(PCA

) and

ISO

(YEA

) by

Dire

ct P

latin

g (C

FU/m

L) a

nd M

embr

ane

Filtr

atio

n (C

FU/1

00m

L)

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Dire

ct P

latin

gM

embr

ane

Filtr

atio

n

Avg.

AQ

HC

(36° C

)

Log

Avg.

AQ

HC

(36° C

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

Avg.

YEA

(3

6° C)

Log

Avg.

YE

A (3

6° C)

Avg.

AQ

HC

(22° C

)

Log

Avg.

AQ

HC

(22° C

)Av

g. Y

EA

(22° C

)

Log

Avg.

YE

A (2

2° C)

Avg.

AQH

C (3

6° C)

(SM

EWW

)

Log

Avg.

AQH

C (3

6° C)

(SM

EWW

)Av

g. P

CA

(36° C

)

Log

Avg.

PC

A (3

6° C)

(SM

EWW

)

Natu

ral

2Ap

ollin

aris

10^

00

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

2Ap

ollin

aris

10^

-10

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

2Ap

ollin

aris

10^

-20

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

E. fa

eciu

m

1Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Ap

ollin

aris

Low

491.

6879

481.

6812

551.

7342

441.

6430

531.

7240

581.

7593

571.

7519

1Ap

ollin

aris

High

120

2.07

8011

62.

0643

119

2.07

4011

22.

0472

114

2.05

4911

62.

0637

125

2.09

51

P. ae

rugi

nosa

2Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Ap

ollin

aris

Low

271.

4287

241.

3662

261.

3920

221.

3424

271.

4287

331.

5085

231.

3495

2Ap

ollin

aris

High

691.

8354

621.

7887

641.

8017

621.

7887

631.

7927

741.

8658

821.

9108

Natu

ral

1Pe

rrier

10^

00

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

1Pe

rrier

10^

-10

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

1Pe

rrier

10^

-20

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

Natu

ral

2Pe

rrier

10^

00

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

2Pe

rrier

10^

-10

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

2Pe

rrier

10^

-20

0.00

00

0.00

00

0.00

00

0.00

00

0.00

0NT

-NT

-

E. c

oli

1Pe

rrier

Unin

ocul

ated

00

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

1Pe

rrier

Low

4747

491.

6855

431.

6305

491.

6869

421.

6232

461.

6574

471.

6668

1Pe

rrier

High

9090

821.

9137

901.

9542

851.

9287

831.

9190

791.

8944

731.

8630

P. ae

rugi

nosa

2Pe

rrier

Unin

ocul

ated

00

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

00.

0000

2Pe

rrier

Low

3030

331.

5106

221.

3295

261.

4024

261.

4058

351.

5322

391.

5898

2Pe

rrier

High

6464

741.

8658

611.

7851

551.

7355

561.

7479

861.

9319

831.

9186

Tabl

e 3:

Res

ults

of A

QCC

vs. S

MEW

W (C

FU/1

00m

L) a

nd IS

O (C

FU/2

50m

L) b

y M

embr

ane

Filtr

atio

n

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Avg.

AQC

C*(S

MEW

W)

Log

Avg.

AQC

C(S

MEW

W)

Avg.

end

o LE

S(S

MEW

W)

Log

Avg.

end

o LE

S(S

MEW

W)

Avg.

AQC

C *

(ISO)

Log

Avg.

AQC

C(IS

O)Av

g. L

acto

se T

TC(IS

O)

Log

Avg.

Lac

tose

TTC

(ISO)

K. p

neum

onia

1Da

sani

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Da

sani

Low

331.

0385

391.

5851

401.

3164

431.

6276

1Da

sani

High

861.

2027

781.

8918

771.

4082

821.

9130

E. c

oli

2Da

sani

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Da

sani

Low

240.

7953

221.

3383

240.

8593

201.

2864

2Da

sani

High

480.

9089

551.

7342

541.

3415

451.

6522

E. c

oli

1Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ne

stle

Pur

elife

Low

191.

2593

161.

1846

181.

2414

221.

3351

1Ne

stle

Pur

elife

High

551.

7349

521.

7118

511.

7016

471.

6668

K. p

neum

onia

2Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ne

stle

Pur

elife

Low

401.

2976

451.

6493

441.

0308

451.

6522

2Ne

stle

Pur

elife

High

761.

5154

831.

9122

731.

2661

791.

8975

E. a

erog

enes

1Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Aq

uafin

aLo

w16

0.72

7911

1.01

6716

0.65

5813

1.06

68

1Aq

uafin

aHi

gh58

1.39

5850

1.69

8651

0.99

5656

1.74

34

E. c

oli

2Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Aq

uafin

aLo

w20

1.04

8723

1.34

6913

0.78

2118

1.24

92

2Aq

uafin

aHi

gh60

1.15

8553

1.71

9268

1.37

0062

1.79

15

E. a

erog

enes

1Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ic

e M

ount

ain

Low

210.

8866

241.

3706

220.

9283

261.

4024

1Ic

e M

ount

ain

High

611.

2829

641.

8049

581.

5377

561.

7465

K. p

neum

onia

2Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ic

e M

ount

ain

Low

411.

2789

451.

6470

481.

6755

361.

5467

2Ic

e M

ount

ain

High

821.

9110

841.

9182

901.

9507

951.

9751

K. p

neum

onia

1Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Kr

oger

Tre

ated

Low

511.

4385

451.

6515

461.

6602

511.

7031

1Kr

oger

Tre

ated

High

861.

3936

821.

9105

841.

1609

931.

9679

E. a

erog

enes

2Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Kr

oger

Tre

ated

Low

260.

9372

301.

4515

310.

9169

261.

4137

2Kr

oger

Tre

ated

High

671.

8256

621.

7910

641.

8017

581.

7634

E. c

oli

1Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Tr

auth

Tre

ated

Low

271.

0372

271.

4169

161.

1963

131.

1021

1Tr

auth

Tre

ated

High

621.

2838

571.

7553

521.

0175

471.

6657

E. a

erog

enes

2Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Tr

auth

Tre

ated

Low

211.

0059

191.

2657

261.

1078

341.

5238

2Tr

auth

Tre

ated

High

701.

3613

651.

8080

541.

2068

691.

8354

*Sum

of T

ypic

al a

nd A

typi

cal c

olon

ies

Tabl

e 3

(con

tinue

d): R

esul

ts o

f AQC

C vs

. SM

EWW

(CFU

/100

mL)

and

ISO

(CFU

/250

mL)

by

Mem

bran

e Fi

ltrat

ion

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Avg.

AQC

C*(S

MEW

W)

Log

Avg.

AQC

C(S

MEW

W)

Avg.

end

o LE

S(S

MEW

W)

Log

Avg.

end

o LE

S(S

MEW

W)

Avg.

AQC

C *

(ISO)

Log

Avg.

AQC

C(IS

O)Av

g. L

acto

se T

TC(IS

O)

Log

Avg.

Lac

tose

TTC

(ISO)

E. a

erog

enes

1Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ev

ian

Low

311.

1405

301.

4749

260.

9234

251.

3869

1Ev

ian

High

661.

1200

611.

7814

671.

5149

621.

7910

E. c

oli

2Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ev

ian

Low

231.

3469

261.

3869

251.

2124

181.

2492

2Ev

ian

High

491.

3066

561.

7441

541.

7260

581.

7549

E. a

erog

enes

1Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Ja

naLo

w22

0.82

5417

1.22

7426

1.11

9823

1.35

12

1Ja

naHi

gh61

1.44

5458

1.75

8960

1.37

3750

1.69

82

E. c

oli

2Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Ja

naLo

w29

1.14

5324

1.37

6824

1.37

8725

1.38

17

2Ja

naHi

gh55

1.31

0254

1.73

2453

1.72

2351

1.70

07

E. c

oli

1Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Fi

jiLo

w18

0.92

8722

1.32

9521

0.87

3516

1.19

63

1Fi

jiHi

gh58

1.36

3354

1.72

7452

1.36

3956

1.74

65

K. p

neum

onia

2Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Fi

jiLo

w50

1.46

7444

1.63

6242

1.39

6446

1.66

18

2Fi

jiHi

gh85

1.46

6385

1.92

6882

1.46

9187

1.93

91

E. c

oli

1Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Vo

ssLo

w28

1.19

7226

1.39

9729

1.06

4429

1.46

14

1Vo

ssHi

gh65

1.50

5858

1.75

8964

1.80

1759

1.76

45

K. p

neum

onia

2Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Vo

ssLo

w44

1.17

2949

1.68

5745

1.61

0848

1.67

89

2Vo

ssHi

gh87

1.25

6378

1.88

9189

1.45

1184

1.92

11

E. c

oli

1Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Vo

ss S

park

ling

Low

321.

1388

281.

4460

321.

0112

281.

4393

1Vo

ss S

park

ling

High

531.

7230

621.

7877

571.

7558

561.

7465

K. p

neum

onia

2Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Vo

ss S

park

ling

Low

431.

1924

471.

6657

421.

2990

361.

5467

2Vo

ss S

park

ling

High

861.

5374

801.

9003

911.

9588

831.

9179

K. p

neum

onia

1Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Ty

Nan

tLo

w49

1.68

9848

1.66

9642

1.31

9748

1.67

55

1Ty

Nan

tHi

gh85

1.71

8777

1.88

3496

1.59

1790

1.95

33

E. a

erog

enes

2Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Ty

Nan

tLo

w34

1.16

4230

1.47

6927

1.22

1523

1.35

51

2Ty

Nan

tHi

gh63

1.16

7460

1.77

4466

1.12

3670

1.84

19

*Sum

of T

ypic

al a

nd A

typi

cal c

olon

ies

Tabl

e 3

(con

tinue

d): R

esul

ts o

f AQC

C vs

. SM

EWW

(CFU

/100

mL)

and

ISO

(CFU

/250

mL)

by

Mem

bran

e Fi

ltrat

ion

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Avg.

AQC

C*(S

MEW

W)

Log

Avg.

AQC

C(S

MEW

W)

Avg.

end

o LE

S(S

MEW

W)

Log

Avg.

end

o LE

S(S

MEW

W)

Avg.

AQC

C *

(ISO)

Log

Avg.

AQC

C(IS

O)Av

g. L

acto

se T

TC(IS

O)

Log

Avg.

Lac

tose

TTC

(ISO)

E. a

erog

enes

1Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Ge

rols

tein

erLo

w30

0.98

2736

1.54

5023

0.90

0828

1.44

47

1Ge

rols

tein

erHi

gh64

1.56

4255

1.73

9263

1.79

8872

1.85

40

K. p

neum

onia

2Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Ge

rols

tein

erLo

w53

1.24

5047

1.66

7251

1.07

8951

1.70

68

2Ge

rols

tein

erHi

gh88

1.26

0093

1.96

8382

1.91

1087

1.93

94

E. a

erog

enes

1Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ap

ollin

aris

Low

271.

0872

221.

3295

220.

9213

261.

4024

1Ap

ollin

aris

High

671.

1841

621.

7915

581.

3779

611.

7839

K. p

neum

onia

2Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ap

ollin

aris

Low

441.

2530

411.

6123

460.

8965

531.

7223

2Ap

ollin

aris

High

911.

5575

841.

9213

911.

3535

751.

8719

E. c

oli

1Pe

rrier

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Pe

rrier

Low

290.

9631

301.

4553

221.

0328

231.

3433

1Pe

rrier

High

541.

7305

561.

7471

671.

8218

631.

7949

E. a

erog

enes

2Pe

rrier

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Pe

rrier

Low

190.

9793

221.

3295

281.

4358

231.

3512

2Pe

rrier

High

560.

9855

491.

6901

531.

7186

541.

7317

*Sum

of T

ypic

al a

nd A

typi

cal c

olon

ies

Tabl

e 4:

Res

ults

of A

QEB

vs. C

usto

mer

Met

hod

(CFU

/100

mL)

by

Mem

bran

e Fi

ltrat

ion

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

AQEB

Ave

rage

Log

AQEB

Aver

age

VRBG

Aver

age

Log

VRBG

Aver

age

K. p

neum

onia

1Da

sani

Unin

ocul

ated

00.

0000

00.

0000

1Da

sani

Low

80.

8891

121.

0603

1Da

sani

High

581.

7628

511.

7031

E. a

erog

enes

2Da

sani

Unin

ocul

ated

00.

0000

00.

0000

2Da

sani

Low

251.

3869

221.

3295

2Da

sani

High

731.

8619

781.

8908

K. p

neum

onia

1Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

1Ne

stle

Pur

elife

Low

50.

6901

30.

4771

1Ne

stle

Pur

elife

High

561.

7413

621.

7885

E. c

oli

2Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

2Ne

stle

Pur

elife

Low

341.

5298

391.

5875

2Ne

stle

Pur

elife

High

100

1.99

6210

62.

0248

Tabl

e 4

(con

tinue

d):

Resu

lts o

f AQE

B vs

. Cus

tom

er M

etho

d (C

FU/1

00m

L) b

y M

embr

ane

Filtr

atio

n

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

AQEB

Ave

rage

Log

AQEB

Aver

age

VRBG

Aver

age

Log

VRBG

Aver

age

E. a

erog

enes

1Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

00

1Aq

uafin

aLo

w40

1.59

9934

1.53

07

1Aq

uafin

aHi

gh95

1.97

7387

1.93

70

K. p

neum

onia

2Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

00

2Aq

uafin

aLo

w13

1.09

6611

1.01

06

2Aq

uafin

aHi

gh54

1.73

1763

1.79

55

E. c

oli

1Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

1Ic

e M

ount

ain

Low

331.

5094

241.

3630

1Ic

e M

ount

ain

High

921.

9634

851.

9259

K. p

neum

onia

2Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

2Ic

e M

ount

ain

Low

60.

7236

40.

5000

2Ic

e M

ount

ain

High

641.

8011

561.

7465

E. c

oli

1Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

1Kr

oger

Tre

ated

Low

521.

7101

481.

6765

1Kr

oger

Tre

ated

High

831.

9179

871.

9393

K. p

neum

onia

2Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

2Kr

oger

Tre

ated

Low

131.

0923

111.

0396

2Kr

oger

Tre

ated

High

651.

8073

551.

7342

E. a

erog

enes

1Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

1Tr

auth

Tre

ated

Low

100.

9911

131.

1021

1Tr

auth

Tre

ated

High

531.

7186

571.

7507

E. c

oli

2Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

2Tr

auth

Tre

ated

Low

311.

4856

361.

5502

2Tr

auth

Tre

ated

High

701.

8444

761.

8772

K. p

neum

onia

1Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

1Ev

ian

Low

40.

3891

60.

7526

1Ev

ian

High

531.

7186

571.

7516

E. a

erog

enes

2Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

2Ev

ian

Low

211.

3095

261.

4098

2Ev

ian

High

771.

8852

741.

8655

E. c

oli

1Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

00

1Ja

naLo

w53

1.72

4053

1.72

23

1Ja

naHi

gh98

1.98

7488

1.94

40

K. p

neum

onia

2Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

00

2Ja

naLo

w7

0.75

935

0.69

01

2Ja

naHi

gh60

1.77

6653

1.72

00

Tabl

e 4

(con

tinue

d):

Resu

lts o

f AQE

B vs

. Cus

tom

er M

etho

d (C

FU/1

00m

L) b

y M

embr

ane

Filtr

atio

n

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

AQEB

Ave

rage

Log

AQEB

Aver

age

VRBG

Aver

age

Log

VRBG

Aver

age

E. c

oli

1Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

00

1Fi

jiLo

w44

1.63

7941

1.60

72

1Fi

jiHi

gh91

1.95

5881

1.90

54

E. a

erog

enes

2Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

00

2Fi

jiLo

w28

1.44

2726

1.41

50

2Fi

jiHi

gh98

1.98

8791

1.95

84

E. c

oli

1Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

00

1Vo

ssLo

w30

1.46

2125

1.38

91

1Vo

ssHi

gh75

1.86

8679

1.89

48

E. a

erog

enes

2Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

00

2Vo

ssLo

w22

1.32

9526

1.40

24

2Vo

ssHi

gh78

1.88

8969

1.83

81

E. c

oli

1Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

1Vo

ss S

park

ling

Low

311.

4905

341.

5298

1Vo

ss S

park

ling

High

731.

8627

801.

9019

E. a

erog

enes

2Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

2Vo

ss S

park

ling

Low

291.

4532

221.

3406

2Vo

ss S

park

ling

High

921.

9634

821.

9137

E. a

erog

enes

1Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

00

1Ty

Nan

tLo

w24

1.36

8618

1.25

46

1Ty

Nan

tHi

gh62

1.79

1569

1.83

56

E. c

oli

2Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

00

2Ty

Nan

tLo

w52

1.70

9342

1.62

28

2Ty

Nan

tHi

gh99

1.99

2489

1.94

87

K. p

neum

onia

1Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

00

1Ge

rols

tein

erLo

w12

1.05

029

0.92

25

1Ge

rols

tein

erHi

gh57

1.75

1663

1.79

18

E. c

oli

2Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

00

2Ge

rols

tein

erLo

w38

1.57

3744

1.64

24

2Ge

rols

tein

erHi

gh91

1.95

8889

1.94

91

K. p

neum

onia

1Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

1Ap

ollin

aris

Low

20.

1505

40.

6021

1Ap

ollin

aris

High

681.

8313

621.

7885

E. a

erog

enes

2Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

2Ap

ollin

aris

Low

221.

3227

281.

4358

2Ap

ollin

aris

High

711.

8491

661.

8147

Tabl

e 4

(con

tinue

d):

Resu

lts o

f AQE

B vs

. Cus

tom

er M

etho

d (C

FU/1

00m

L) b

y M

embr

ane

Filtr

atio

n

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

AQEB

Ave

rage

Log

AQEB

Aver

age

VRBG

Aver

age

Log

VRBG

Aver

age

K.

pneu

mon

ia

1Pe

rrie

rUn

inoc

ulat

ed0

0.00

000

0.00

00

1Pe

rrie

rLo

w13

1.11

2714

1.13

59

1Pe

rrie

rHi

gh53

1.71

9261

1.78

32

E. a

erog

enes

2Pe

rrie

rUn

inoc

ulat

ed0

0.00

000

0.00

00

2Pe

rrie

rLo

w22

1.32

9524

1.37

68

2Pe

rrie

rHi

gh82

1.91

0286

1.93

11

Tabl

e 5:

Res

ults

of E

C vs

. SM

EWW

(CFU

/100

mL)

and

ISO

(CFU

/250

mL)

by

Mem

bran

e Fi

ltrat

ion:

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Avg.

EC*

(SM

EWW

)

Log

Avg.

EC

(SM

EWW

)Av

g. e

ndo

LES

(SM

EWW

)

Log

Avg.

end

o LE

S(S

MEW

W)

Avg.

EC*

(ISO)

Log

Avg.

EC

(ISO)

Avg.

Lac

tose

TTC

(ISO)

Log

Avg.

Lac

tose

TTC

(ISO)

K. p

neum

onia

1Da

sani

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Da

sani

Low

401.

6334

481.

6809

330.

2401

470.

9358

1Da

sani

High

821.

9345

881.

9417

730.

2853

821.

1105

E. c

oli

2Da

sani

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Da

sani

Low

301.

5185

221.

3406

250.

2232

340.

8325

2Da

sani

High

681.

8513

621.

7885

820.

2911

861.

1255

E. c

oli

1Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ne

stle

Pur

elife

Low

281.

3010

221.

3295

230.

2165

280.

7601

1Ne

stle

Pur

elife

High

721.

8195

561.

7434

750.

2863

751.

0879

K. p

neum

onia

2Ne

stle

Pur

elife

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ne

stle

Pur

elife

Low

451.

4776

381.

5792

350.

2438

260.

8137

2Ne

stle

Pur

elife

High

821.

9345

711.

8473

700.

2831

781.

0776

E. a

erog

enes

1Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Aq

uafin

aLo

w18

1.34

2413

1.08

8021

0.20

9221

0.66

03

1Aq

uafin

aHi

gh60

1.80

6253

1.72

3053

0.26

8048

0.94

18

E. c

oli

2Aq

uafin

aUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Aq

uafin

aLo

w36

1.50

5132

1.49

7830

0.23

5220

0.69

28

2Aq

uafin

aHi

gh57

1.81

2964

1.80

1782

0.29

1474

1.05

17

E. a

erog

enes

1Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ic

e M

ount

ain

Low

201.

2304

201.

2870

250.

2232

320.

8233

1Ic

e M

ount

ain

High

611.

7559

701.

8419

630.

2779

671.

0315

K. p

neum

onia

2Ic

e M

ount

ain

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ic

e M

ount

ain

Low

541.

7403

461.

6623

400.

2527

350.

8742

2Ic

e M

ount

ain

High

901.

9345

951.

9749

740.

2859

831.

1105

*Sum

of T

ypic

al a

nd A

typi

cal c

olon

ies

Tabl

e 5

(con

tinue

d):

Resu

lts o

f EC

vs. S

MEW

W (C

FU/1

00m

L) a

nd IS

O (C

FU/2

50m

L) b

y M

embr

ane

Filtr

atio

n:

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Avg.

EC*

(SM

EWW

)

Log

Avg.

EC

(SM

EWW

)Av

g. e

ndo

LES

(SM

EWW

)

Log

Avg.

end

o LE

S(S

MEW

W)

Avg.

EC*

(ISO)

Log

Avg.

EC

(ISO)

Avg.

Lac

tose

TTC

(ISO)

Log

Avg.

Lac

tose

TTC

(ISO)

K. p

neum

onia

1Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Kr

oger

Tre

ated

Low

441.

5412

381.

5709

420.

2550

530.

9954

1Kr

oger

Tre

ated

High

831.

8741

771.

8816

710.

2842

651.

0272

E. a

erog

enes

2Kr

oger

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Kr

oger

Tre

ated

Low

231.

4311

171.

2274

290.

2307

240.

7218

2Kr

oger

Tre

ated

High

521.

7559

641.

8024

640.

2784

721.

0594

E. c

oli

1Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Tr

auth

Tre

ated

Low

231.

4314

271.

4302

300.

2341

290.

7601

1Tr

auth

Tre

ated

High

821.

8976

701.

8440

690.

2828

651.

0517

E. a

erog

enes

2Tr

auth

Tre

ated

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Tr

auth

Tre

ated

Low

251.

3424

201.

3005

290.

2311

230.

7829

2Tr

auth

Tre

ated

High

791.

9345

601.

7766

890.

2951

831.

1105

E. a

erog

enes

1Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ev

ian

Low

261.

5315

231.

3512

380.

2488

360.

8742

1Ev

ian

High

671.

8808

651.

8085

830.

2916

881.

1255

E. c

oli

2Ev

ian

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ev

ian

Low

351.

5911

261.

4147

270.

2272

300.

8325

2Ev

ian

High

751.

8062

851.

9265

740.

2863

861.

0946

E. a

erog

enes

1Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Ja

naLo

w25

1.39

7920

1.28

6424

0.21

9931

0.80

38

1Ja

naHi

gh67

1.87

5161

1.78

4876

0.28

7368

1.07

04

E. c

oli

2Ja

naUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Ja

naLo

w25

1.34

2431

1.48

5630

0.23

4322

0.72

18

2Ja

naHi

gh63

1.85

1368

1.82

9276

0.28

7678

1.05

94

E. c

oli

1Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Fi

jiLo

w34

1.55

6332

1.50

3227

0.22

8216

0.69

28

1Fi

jiHi

gh80

1.86

9265

1.80

8572

0.28

4579

1.07

04

K. p

neum

onia

2Fi

jiUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Fi

jiLo

w36

1.55

6346

1.66

1839

0.25

0637

0.89

64

2Fi

jiHi

gh86

1.95

9083

1.91

6182

0.29

1378

1.07

40

E. c

oli

1Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Vo

ssLo

w38

1.61

2832

1.49

6925

0.22

2433

0.85

83

1Vo

ssHi

gh63

1.77

8271

1.85

0972

0.28

4983

1.11

05

K. p

neum

onia

2Vo

ssUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Vo

ssLo

w47

1.43

1137

1.22

7448

0.23

0743

0.72

18

2Vo

ssHi

gh65

1.75

5980

1.80

2485

0.27

8480

1.05

94

*Sum

of T

ypic

al a

nd A

typi

cal c

olon

ies

Tabl

e 5

(con

tinue

d):

Resu

lts o

f EC

vs. S

MEW

W (C

FU/1

00m

L) a

nd IS

O (C

FU/2

50m

L) b

y M

embr

ane

Filtr

atio

n:

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um L

evel

Avg.

EC*

(SM

EWW

)

Log

Avg.

EC

(SM

EWW

)Av

g. e

ndo

LES

(SM

EWW

)

Log

Avg.

end

o LE

S(S

MEW

W)

Avg.

EC*

(ISO)

Log

Avg.

EC

(ISO)

Avg.

Lac

tose

TTC

(ISO)

Log

Avg.

Lac

tose

TTC

(ISO)

E. c

oli

1Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Vo

ss S

park

ling

Low

351.

5911

351.

5680

330.

2627

280.

9102

1Vo

ss S

park

ling

High

741.

7853

841.

8993

800.

2931

711.

1105

K. p

neum

onia

2Vo

ss S

park

ling

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Vo

ss S

park

ling

Low

381.

5185

361.

5376

400.

2401

480.

8137

2Vo

ss S

park

ling

High

731.

8976

561.

9213

820.

2902

801.

0478

K. p

neum

onia

1Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Ty

Nan

tLo

w41

1.55

5644

1.55

2138

0.25

2846

0.97

00

1Ty

Nan

tHi

gh83

1.77

6674

1.74

1366

0.29

1480

1.10

74

E. a

erog

enes

2Ty

Nan

tUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Ty

Nan

tLo

w31

1.66

2833

1.63

6215

0.24

9829

0.92

33

2Ty

Nan

tHi

gh62

1.90

3172

1.86

6074

0.28

0474

1.07

76

E. a

erog

enes

1Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Ge

rols

tein

erLo

w26

1.54

4126

1.50

8522

0.18

4122

0.78

29

1Ge

rols

tein

erHi

gh65

1.82

6154

1.85

3873

0.28

5666

1.08

11

K. p

neum

onia

2Ge

rols

tein

erUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Ge

rols

tein

erLo

w41

1.43

0253

1.40

2433

0.21

4225

0.66

03

2Ge

rols

tein

erHi

gh83

1.77

8294

1.73

0563

0.28

5353

1.03

15

E. a

erog

enes

1Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

1Ap

ollin

aris

Low

201.

6180

131.

7240

240.

2399

220.

7352

1Ap

ollin

aris

High

611.

8692

641.

9727

810.

2780

730.

9954

K. p

neum

onia

2Ap

ollin

aris

Unin

ocul

ated

00.

0000

00.

0000

00.

0000

00.

0000

2Ap

ollin

aris

Low

501.

2041

411.

0880

460.

2199

490.

7601

2Ap

ollin

aris

High

811.

7076

761.

8021

740.

2904

751.

0478

E. c

oli

1Pe

rrie

rUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

1Pe

rrie

rLo

w24

1.66

2830

1.60

5834

0.26

0234

0.94

18

1Pe

rrie

rHi

gh80

1.92

9467

1.87

7579

0.28

5975

1.09

46

E. a

erog

enes

2Pe

rrie

rUn

inoc

ulat

ed0

0.00

000

0.00

000

0.00

000

0.00

00

2Pe

rrie

rLo

w24

1.44

7229

1.47

6934

0.24

2040

0.86

64

2Pe

rrie

rHi

gh68

1.86

9263

1.82

5960

0.28

9270

1.05

94

*Sum

of T

ypic

al a

nd A

typi

cal c

olon

ies

Tabl

e 6:

Res

ults

of A

QYM

vs.

SM

EWW

(CRB

) and

Cus

tom

er M

etho

d (a

PDA)

by

Mem

bran

e Fi

ltrat

ion

(CFU

/100

mL)

:

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um

Leve

l

3 Da

y Co

unts

(CFU

/100

mL)

5 Da

y Co

unts

(CFU

/100

mL)

Avg.

AQ

YM

(25° C

)

Log

Avg.

AQ

YM

(25° C

)

Avg.

aP

DA

(25° C

)

Log

Avg.

aP

DA

(25° C

)

Avg.

AQ

YM

(20° C

)

Log

Avg.

AQ

YM(2

0° C)

Avg.

CR

B (2

0° C)

Log

Avg.

CR

B (2

0° C)

Avg.

AQ

YM

(25° C

)

Log

Avg.

AQ

YM

(25° C

)

Avg.

aP

DA

(25° C

)

Log

Avg.

aP

DA

(25° C

)

Avg.

AQ

YM

(20° C

)

Log

Avg.

AQ

YM(2

0° C)

Avg.

CR

B (2

0° C)

Log

Avg.

CR

B (2

0° C)

Peni

cilli

um

1Da

sani

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Da

sani

Low

211.

3106

221.

3406

211.

3106

221.

3266

211.

3106

221.

3406

221.

3424

221.

3266

1Da

sani

High

661.

8161

631.

7952

661.

8187

621.

7915

661.

8161

631.

7952

671.

8256

661.

8162

Paec

ilom

yces

2Da

sani

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Da

sani

Low

271.

4265

281.

4460

301.

4749

301.

4647

311.

4893

301.

4732

311.

4814

311.

4893

2Da

sani

High

831.

9183

851.

9287

771.

8862

841.

9204

841.

9211

851.

9287

801.

9002

851.

9287

Paec

ilom

yces

1Ne

stle

Pur

elife

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Ne

stle

Pur

elife

Low

371.

5603

341.

5298

361.

5563

361.

5536

391.

5909

371.

5656

381.

5740

371.

5603

1Ne

stle

Pur

elife

High

881.

9444

821.

9130

881.

9427

841.

9232

921.

9634

861.

9314

901.

9533

861.

9314

Peni

cilli

um

2Ne

stle

Pur

elife

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Ne

stle

Pur

elife

Low

271.

4302

281.

4401

251.

3869

241.

3706

271.

4302

281.

4401

251.

3869

251.

3923

2Ne

stle

Pur

elife

High

591.

7664

621.

7887

701.

8394

641.

8049

591.

7703

621.

7887

721.

8563

661.

8171

Paec

ilom

yces

1Aq

uafin

aUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Aq

uafin

aLo

w42

1.62

2834

1.52

6736

1.55

5635

1.53

2243

1.62

7636

1.54

6738

1.57

9637

1.56

03

1Aq

uafin

aHi

gh81

1.90

8086

1.93

1481

1.90

8278

1.89

1582

1.91

3388

1.94

3583

1.91

8680

1.90

28

C. a

lbic

ans

2Aq

uafin

aUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Aq

uafin

aLo

w26

1.40

5826

1.40

2424

1.37

9824

1.36

8626

1.40

5826

1.41

2125

1.39

7924

1.37

87

2Aq

uafin

aHi

gh93

1.96

8391

1.95

6592

1.96

1389

1.94

9394

1.97

3092

1.96

1493

1.96

6189

1.94

93

C. a

lbic

ans

1Ic

e M

ount

ain

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Ic

e M

ount

ain

Low

281.

4460

281.

4358

291.

4548

301.

4667

291.

4542

281.

4358

291.

4548

301.

4749

1Ic

e M

ount

ain

High

841.

9213

871.

9393

821.

9137

841.

9242

871.

9364

881.

9418

831.

9164

851.

9268

Peni

cilli

um

2Ic

e M

ount

ain

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Ic

e M

ount

ain

Low

221.

3323

221.

3295

211.

3106

231.

3512

221.

3424

221.

3295

231.

3521

231.

3512

2Ic

e M

ount

ain

High

521.

7153

541.

7312

561.

7471

521.

7108

541.

7323

541.

7312

591.

7708

561.

7481

Paec

ilom

yces

1Kr

oger

Tre

ated

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Kr

oger

Tre

ated

Low

331.

5094

331.

5185

291.

4558

331.

5167

351.

5211

351.

5439

311.

4814

331.

5167

1Kr

oger

Tre

ated

High

751.

8741

781.

8889

791.

8968

761.

8768

771.

9054

771.

8859

811.

9080

811.

9085

C. a

lbic

ans

2Kr

oger

Tre

ated

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Kr

oger

Tre

ated

Low

251.

3869

231.

3613

281.

4375

241.

3787

231.

3965

231.

3613

281.

4460

251.

3884

2Kr

oger

Tre

ated

High

761.

8805

801.

9025

831.

9164

781.

8882

801.

8859

801.

9025

841.

9243

811.

9084

Peni

cilli

um

1Tr

auth

Tre

ated

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Tr

auth

Tre

ated

Low

211.

3177

241.

3702

221.

3424

221.

3420

221.

3406

251.

3891

231.

3521

221.

3420

1Tr

auth

Tre

ated

High

531.

7240

511.

7072

551.

7386

521.

7147

541.

7279

521.

7159

561.

7471

561.

7443

Paec

ilom

yces

2Tr

auth

Tre

ated

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Tr

auth

Tre

ated

Low

311.

4877

321.

4978

361.

5536

341.

5238

371.

5603

351.

5434

381.

5731

341.

5307

2Tr

auth

Tre

ated

High

881.

9417

811.

9058

831.

9188

861.

9311

901.

9517

881.

9440

841.

9215

861.

9311

Tabl

e 6

(con

tinue

d):

Resu

lts o

f AQY

M v

s. S

MEW

W (C

RB) a

nd C

usto

mer

Met

hod

(aPD

A) b

y M

embr

ane

Filtr

atio

n (C

FU/1

00m

L):

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um

Leve

l

3 Da

y Co

unts

(CFU

/100

mL)

5 Da

y Co

unts

(CFU

/100

mL)

Avg.

AQ

YM

(25° C

)

Log

Avg.

AQ

YM

(25° C

)

Avg.

aP

DA

(25° C

)

Log

Avg.

aP

DA

(25° C

)

Avg.

AQ

YM

(20° C

)

Log

Avg.

AQ

YM(2

0° C)

Avg.

CR

B (2

0° C)

Log

Avg.

CR

B (2

0° C)

Avg.

AQ

YM

(25° C

)

Log

Avg.

AQ

YM

(25° C

)

Avg.

aP

DA

(25° C

)

Log

Avg.

aP

DA

(25° C

)

Avg.

AQ

YM

(20° C

)

Log

Avg.

AQ

YM(2

0° C)

Avg.

CR

B (2

0° C)

Log

Avg.

CR

B (2

0° C)

Peni

cilli

um

1Ev

ian

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Ev

ian

Low

291.

4621

261.

4058

271.

4232

281.

4375

291.

4621

271.

4314

271.

4232

281.

4375

1Ev

ian

High

691.

8384

721.

8572

671.

8227

651.

8116

711.

8481

721.

8572

671.

8227

651.

8116

C. a

lbic

ans

2Ev

ian

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Ev

ian

Low

231.

3551

241.

3787

241.

3802

261.

4044

251.

3869

251.

3884

251.

3976

261.

4044

2Ev

ian

High

841.

9204

851.

9265

821.

9138

831.

9188

851.

9259

851.

9265

841.

9242

831.

9188

Paec

ilom

yces

1Ja

naUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Ja

naLo

w32

1.49

5633

1.51

5333

1.51

6732

1.50

4335

1.53

7434

1.52

8534

1.53

0733

1.51

67

1Ja

naHi

gh74

1.86

5582

1.91

3779

1.89

4480

1.90

2881

1.90

5683

1.91

9080

1.90

3081

1.90

56

Peni

cilli

um

2Ja

naUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Ja

naLo

w20

1.29

6119

1.27

8823

1.36

1324

1.37

8722

1.34

0620

1.30

0523

1.36

1324

1.37

87

2Ja

naHi

gh73

1.85

9573

1.86

0165

1.80

9472

1.85

7275

1.87

1973

1.86

0172

1.85

3873

1.86

01

Paec

ilom

yces

1Fi

jiUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Fi

jiLo

w36

1.55

4835

1.53

9634

1.52

6732

1.50

4337

1.56

1339

1.57

7136

1.54

8134

1.52

46

1Fi

jiHi

gh88

1.94

1784

1.92

3888

1.93

9982

1.91

3790

1.95

3386

1.93

1490

1.95

3389

1.94

94

Peni

cilli

um

2Fi

jiUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Fi

jiLo

w22

1.33

1423

1.34

6924

1.37

8726

1.41

3722

1.33

1423

1.35

8026

1.40

6526

1.41

37

2Fi

jiHi

gh79

1.89

4875

1.87

2173

1.86

3271

1.84

7979

1.89

4875

1.87

2174

1.86

6272

1.85

42

C. a

lbic

ans

1Vo

ssUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Vo

ssLo

w24

1.36

8627

1.42

8726

1.40

9823

1.34

6924

1.37

8727

1.42

8727

1.42

8724

1.36

30

1Vo

ssHi

gh82

1.91

3084

1.92

1387

1.93

9382

1.91

3883

1.91

5880

1.90

0387

1.93

9384

1.92

17

Peni

cilli

um

2Vo

ssUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Vo

ssLo

w20

1.29

8820

1.28

2925

1.38

8425

1.38

6920

1.29

8820

1.28

2926

1.40

6525

1.38

69

2Vo

ssHi

gh70

1.84

4466

1.81

4168

1.82

9069

1.83

5471

1.85

0968

1.82

8368

1.83

2369

1.83

54

Paec

ilom

yces

1Vo

ss S

park

ling

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Vo

ss S

park

ling

Low

361.

5561

351.

5374

351.

5396

321.

5049

391.

5845

371.

5613

371.

5668

341.

5246

1Vo

ss S

park

ling

High

821.

9110

841.

9238

831.

9163

821.

9130

841.

9242

841.

9213

841.

9242

851.

9289

C. a

lbic

ans

2Vo

ss S

park

ling

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Vo

ss S

park

ling

Low

321.

4978

331.

5094

311.

4795

331.

5153

321.

4978

331.

5094

321.

4956

331.

5153

2Vo

ss S

park

ling

High

891.

9464

851.

9259

831.

9186

801.

9028

911.

9589

861.

9314

841.

9211

811.

9054

C. a

lbic

ans

1Ty

Nan

tUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Ty

Nan

tLo

w29

1.46

1424

1.37

8726

1.40

6525

1.38

9130

1.47

6925

1.38

8426

1.41

5026

1.40

98

1Ty

Nan

tHi

gh91

1.95

5884

1.92

3887

1.93

6781

1.90

8091

1.95

5884

1.92

3887

1.93

6782

1.91

35

Peni

cilli

um

2Ty

Nan

tUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Ty

Nan

tLo

w24

1.37

8726

1.41

3723

1.35

5123

1.35

8025

1.39

7626

1.41

3725

1.38

6924

1.36

86

2Ty

Nan

tHi

gh70

1.84

4472

1.85

3868

1.83

2571

1.84

8170

1.84

4472

1.85

3870

1.84

2071

1.84

81

Tabl

e 6

(con

tinue

d):

Resu

lts o

f AQY

M v

s. S

MEW

W (C

RB) a

nd C

usto

mer

Met

hod

(aPD

A) b

y M

embr

ane

Filtr

atio

n (C

FU/1

00m

L):

Stra

inLo

tM

atrix

Bra

ndIn

ocul

um

Leve

l

3 Da

y Co

unts

(CFU

/100

mL)

5 Da

y Co

unts

(CFU

/100

mL)

Avg.

AQ

YM

(25° C

)

Log

Avg.

AQ

YM

(25° C

)

Avg.

aP

DA

(25° C

)

Log

Avg.

aP

DA

(25° C

)

Avg.

AQ

YM

(20° C

)

Log

Avg.

AQ

YM(2

0° C)

Avg.

CR

B (2

0° C)

Log

Avg.

CR

B (2

0° C)

Avg.

AQ

YM

(25° C

)

Log

Avg.

AQ

YM

(25° C

)

Avg.

aP

DA

(25° C

)

Log

Avg.

aP

DA

(25° C

)

Avg.

AQ

YM

(20° C

)

Log

Avg.

AQ

YM(2

0° C)

Avg.

CR

B (2

0° C)

Log

Avg.

CR

B (2

0° C)

Peni

cilli

um

1Ge

rols

tein

erUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Ge

rols

tein

erLo

w21

1.32

0223

1.35

2126

1.40

2425

1.39

7623

1.35

2123

1.35

2127

1.42

1326

1.41

50

1Ge

rols

tein

erHi

gh69

1.83

8871

1.84

7975

1.87

1969

1.83

8169

1.83

8871

1.84

7975

1.87

4971

1.85

09

C. a

lbic

ans

2Ge

rols

tein

erUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Ge

rols

tein

erLo

w23

1.36

1325

1.38

8424

1.37

6828

1.44

6024

1.37

1025

1.38

8425

1.38

6928

1.44

60

2Ge

rols

tein

erHi

gh81

1.90

8078

1.89

2086

1.93

1191

1.95

6382

1.91

3579

1.89

4687

1.93

6492

1.96

37

Paec

ilom

yces

1Ap

ollin

aris

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

1Ap

ollin

aris

Low

361.

5548

361.

5498

321.

4969

311.

4828

371.

5676

361.

5556

331.

5183

321.

5049

1Ap

ollin

aris

High

831.

9186

861.

9314

881.

9444

861.

9344

881.

9445

861.

9340

891.

9468

881.

9445

C. a

lbic

ans

2Ap

ollin

aris

Unin

ocul

ated

0NA

0NA

0NA

0NA

0NA

0NA

0NA

0NA

2Ap

ollin

aris

Low

261.

4147

261.

4098

271.

4287

231.

3521

271.

4225

271.

4287

291.

4542

281.

4387

2Ap

ollin

aris

High

801.

9025

831.

9155

841.

9235

771.

8837

811.

9051

841.

9204

851.

9289

791.

8976

C. a

lbic

ans

1Pe

rrie

rUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

1Pe

rrie

rLo

w26

1.41

3728

1.43

7522

1.33

1424

1.37

0627

1.43

0228

1.43

7524

1.37

1025

1.38

17

1Pe

rrie

rHi

gh91

1.95

6385

1.92

5983

1.91

6185

1.92

6892

1.96

1386

1.93

4085

1.92

6885

1.92

68

Paec

ilom

yces

2Pe

rrie

rUn

inoc

ulat

ed0

NA0

NA0

NA0

NA0

NA0

NA0

NA0

NA

2Pe

rrie

rLo

w35

1.54

1239

1.58

5136

1.55

6334

1.52

6736

1.54

8139

1.58

5138

1.57

9236

1.54

81

2Pe

rrie

rHi

gh91

1.95

6386

1.93

4091

1.95

8694

1.97

2792

1.96

1187

1.93

9392

1.96

0995

1.97

51

Tabl

e 7:

Com

paris

on o

f the

3M

Pet

rifilm

Aqu

a Pl

ate

met

hod

& co

rresp

ondi

ng re

fere

nce

met

hod

for t

he e

num

erat

ion

of b

acte

ria in

ocul

ated

in b

ottle

d w

ater

Bact

eria

Com

paris

onLe

vel

NM

LDSE

MT

p-va

lue

Repe

atab

ility

Petr

ifilm

Refe

renc

e

Hete

rotr

ophi

cs

AQHC

Pla

te to

PCA

(36o C

) (di

rect

)Un

inoc

100.

160.

022

0.72

0.48

80.

060@

0.10

0

AQHC

Pla

te to

PCA

(36o C

) (di

rect

)Lo

w30

0.00

00.

011

0.02

0.98

50.

065

0.05

5

AQHC

Pla

te to

PCA

(36o C

) (di

rect

)Hi

gh30

0.00

10.

010

0.10

0.91

70.

038

0.03

0

AQHC

Pla

te to

YEA

(36o C

) (di

rect

)Un

inoc

10-0

.001

0.01

4-0

.06

0.95

4NA

NA

AQHC

Pla

te to

YEA

(36o C

) (di

rect

)Lo

w30

-0.0

070.

013

-0.5

40.

593

NANA

AQHC

Pla

te to

YEA

(36o C

) (di

rect

)Hi

gh30

0.01

10.

006

1.79

0.08

4NA

NA

AQHC

Pla

te to

YEA

(22o C

) (di

rect

)Un

inoc

100.

029

0.03

80.

770.

463

NANA

AQHC

Pla

te to

YEA

(22o C

) (di

rect

)Lo

w30

-0.0

030.

008

-0.4

10.

688

NANA

AQHC

Pla

te to

YEA

(22o C

) (di

rect

)Hi

gh30

0.00

60.

005

1.30

0.20

2NA

NA

AQHC

Pla

te to

PCA

(36o C

) (fil

tere

d)Un

inco

90.

029

0.02

21.

350.

212

0.04

80.

063

AQHC

Pla

te to

PCA

(36o C

) (fil

tere

d)Lo

w30

-0.0

020.

011

-0.2

10.

833

0.06

00.

053

AQHC

Pla

te to

PCA

(36o C

) (fil

tere

d)Hi

gh30

-0.0

030.

006

-0.5

30.

603

0.02

60.

032

Colif

orm

AQCC

Pla

te to

end

o LE

S (S

MEW

W)

Low

300.

010

0.01

20.

850.

403

0.06

9@0.

096

AQCC

Pla

te to

end

o LE

S (S

MEW

W)

High

300.

017

0.00

72.

560.

016*

0.04

00.

039

AQCC

Pla

te to

Lac

tose

TTC

(ISO

)Lo

w30

0.01

00.

014

0.69

0.49

50.

062@

0.09

4

AQCC

Pla

te to

Lac

tose

TTC

(ISO

)Hi

gh30

0.00

40.

008

0.48

0.63

80.

038

0.05

0

Ente

roba

cter

iace

aeAQ

EB P

late

to V

RBG

Low

30-0

.005

0.02

7-0

.20

0.84

40.

161

0.10

7@

AQEB

Pla

te to

VRB

GHi

gh30

0.00

60.

008

0.73

0.47

0.04

80.

038

Yeas

t & M

old

AQYM

Pla

te to

CRB

(20o C

) (3

Day)

Low

300.

002

0.00

60.

330.

745

0.06

00.

072

AQYM

Pla

te to

CRB

(20o C

) (3

Day)

High

300.

008

0.00

41.

930.

063

0.03

20.

031

AQYM

Pla

te to

PDA

(25o C

) (3

Day)

Low

30-0

.001

0.00

6-0

.20

0.84

40.

058

0.06

3

AQYM

Pla

te to

PDA

(25o C

) (3

Day)

High

300.

001

0.00

40.

360.

723

0.03

00.

033

AQYM

Pla

te to

CRB

(20o C

) (5

Day)

Low

300.

007

0.00

51.

600.

121

0.04

1@0.

063

AQYM

Pla

te to

CRB

(20o C

) (5

Day)

High

300.

006

0.00

32.

030.

052

0.02

50.

024

AQYM

Pla

te to

PDA

(25o C

) (5

Day)

Low

300.

004

0.00

60.

670.

506

0.04

4@0.

063

AQYM

Pla

te to

PDA

(25o C

) (5

Day)

High

300.

007

0.00

32.

200.

036*

0.02

60.

028

NNu

mbe

r of s

ampl

es w

ith c

ount

able

resu

lts

MLD

Mea

n lo

g di

ffere

nce

coun

t per

sam

ple

(log

3M P

etrifi

lm P

late

– lo

g re

fere

nce

met

hod)

SEM

Stan

dard

err

or o

f the

mea

n lo

g di

ffere

nce

TCa

lcul

ated

t-st

atis

tic

p-va

lue

Actu

al s

igni

fican

ce le

vel o

f the

cal

cula

ted

stat

istic

Repe

atab

ility

Clos

enes

s of

agr

eem

ent b

etw

een

succ

essi

ve re

sults

obt

aine

d w

ith th

e sa

me

met

hod

on id

entic

al te

st m

ater

ial a

nd u

nder

the

sam

e co

nditi

ons

and

is e

xpre

ssed

as

a st

anda

rd d

evia

tion

*Si

gnifi

cant

ly d

iffer

ent (

p-va

lue<

0.05

)

@Si

gnifi

cant

ly b

ette

r rep

eata

bilit

y (p

<0.

05)

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