# ATP Case No. N09-004 Study Report

1 

# ATP Study Report for the Prop osed US EPA Approval of

Colilert ® 

# and Colilert ® 

# -18 for the Detection and Enumeration of

Fecal Coliforms in US Wastewater Samples

February 1, 2010

IDEXX Laboratories, Inc.

Westbrook, Maine 04092

# ATP Case No. N09-004 Study Report

2 Background

Introduction:

This report is intended to review the dat a collected to compar e the performance

of the Colilert and Colilert-18 test methods against the Standard Method (i.e.

SM9222D) from Standard Methods for the Examin ation of Water and Wastewater

for detecting fecal coliform bacteria in wastewater samples ( Appendix D , 3 ).

Colilert and Colilert-18 are US EPA approved for t he detection and quantification

of total coliforms & Escherichia coli from source and drinking water samples.

Both methods are also approved for the specific detection and quantification of E.

coli from wastewater samples. This repor t describes a study that was initiated

following the US EPA 2004 Alternative Test Procedure (ATP) guidance protocol

( Appendix D , 1 ) to evaluate the performance of both methods at detecting and

quantifying fecal coliform bacteria in s upport of a US EPA approval initiative.

Both Colilert and Coliler t-18 are manufactured by IDEXX Laboratories, inc.

located in Westbrook, Maine ( Appendix A(1) and Appendix A(2) )

Data generated for this study were colle cted in accordance with the ATP Study

Plan approved by the US EPA on July 21 st 

# , 2009 (Appendix B).

Justification:

Colilert and Colilert-18 can detect and quantify the conc entration of total coliform

& E. coli bacteria from potable water sa mples and the concentration of E. coli

from wastewater samples after 24 and 18 hours, respectively. Both methods

offer considerable advantages to laborat ory analysts in terms of their ease of

use, accuracy, and time to results ov er SM9222D and have been widely adopted

in most public and private laboratories fo r US EPA mandated testing. Although

40 CFR 136.3, Table IA—List of Approv ed Biological Methods for Wastewater

and Sewage Sludge ( Appendix D , 2 ) , allows for the use of E coli as one of a list

of allowable target organi sms under the rule, some st ates have not yet approved

the use of E. coli monitoring of wastewater disc harge and, as a result, can not

use Colilert and Colilert-18 for wastewater monitoring. Therefore, IDEXX initiated

this study to obtain US EPA approval for fecal coliform testing for both Colilert

and Colilert-18 to allow these states access to these methods.

Method Summary:

The Colilert and Colilert-18 tests are manufactured in snap packs; one snap pack

contains enough powder to perform one test for the presence of fecal coliform

bacteria in 100mL of wastewater. The user simply adds the contents of one of

the snap pack to a vessel containing 100mL of a wastewater sample followed by

shaking to quickly dissolve the powder into the liquid reagent. After shaking, the

reagent is transferred to a sterile Quanti-Tray device (Quanti-Tray or QuantiTray/2000 from IDEXX Laboratories, Inc.) which dispenses and seals the reagent

into a large number of incubating wells. The Quanti-Tray is then incubated at

44.5 +/-0.2 ° C for 24 hours (when using Colilert) or 18 hours (when using Colilert18). After incubation, the number of wells which have changed color to yellow

are counted and compared against a most probable number (MPN) chart

# ATP Case No. N09-004 Study Report

3 (provided with the Quanti-Trays) to arrive at the final number of fecal coliform

bacteria detected in the origin al wastewater sample.

Equivocal samples are compared for co lor change against a color comparator

(IDEXX Laboratories Inc.) In the Study Method, fecal coliform detection is based

on the presence of the enzyme ß-D-galacto sidase which is known to be present

in fecal coliform bacteria ( Appendices A(1) and A(2) ) For the detection of ß-Dgalactosidase, Colilert and Colilert-18 ut ilize the chromogenic nutrient indicator

ortho-nitrophenyl-ß-D-galact opyranoside (ONPG) which produces a distinct

yellow color when hydrolyzed by ß-D-galacto sidase. The presence of this yellow

color under these incubation conditions is highly specific for the detection of fecal

coliform bacteria.

Study Objectives and Design

Main Objectives:

The main objective of this ATP study was to demonstrate the comparability of the

Colilert and Colilert-18 methods to a US EPA approved method (SM9222D) for

compliance monitoring of fecal coliform di scharge from wastewat er facilities.

IDEXX Laboratories, Inc. carried out this AT P study in support of their request for

nationwide approval of Colilert and Colilert-18 by the US EPA.

Data Quality Objectives:

IDEXX coordinated and served as the Study Director for this ATP after approval

of the proposed study plan by the US EPA. The data was collected and recorded

at an independent certified laboratory approved by the US EPA which has

provided a bound copy of all the data collected during this study (Appendix C)

.

Study Design:

The design of this study was based on the US EPA’s Microbiological Alternative

Test Procedure Protocol for Drinking Water, Ambient Water and Wastewater,

version 2004 and is described in more detail in the US EPA approved protocol

(Appendix B) . In summary, primary wastewater effluent samples were collected

from 10 different locations within the New England area and used as sources of

fecal coliform bacteria for this study. T he effluent samples were collected at the

individual wastewater treat ment plants, processed, and tested at the certified

laboratory within six h ours. If necessary, the effl uent samples were diluted to

achieve a fecal coliform innoculum of approximately 20 to 50 organisms per

100mL of sample. Each sample was analyzed by Colilert, Colilert-18, and

SM9222D in replicates of twenty after incubation at 44.5+/-0.2 ° C for the

appropriate period of time. Sensitivity for fecal coliform detection was

determined by comparing the number of presumptive positive responses for the

Colilert and Colilert-18 methods agains t the number of pres umptive positive

responses from SM9222D. Specificity wa s determined by confirming the identity

at least 200 presumptive positive and 200 presumptive negative determinations

from each of the three test methods. These data were used to determine the

statistical comparability of the feca l coliform counts determined by each method

following the ATP guidelines as well as the confirmed false positive and false

negative rates.

# ATP Case No. N09-004 Study Report

4 Study Implementation

Study management:

All laboratory work for this study, except for VITEK iden tifications, was carried out

at Analytical Services, Inc. (ASI) locat ed in Williston, Vermont. IDEXX served as

the Study Director and obtained approval of the Study Plan and th e writing of the

Study Report. ASI was solely responsib le for collecting the effluent samples,

running the experiments, recording t he data, and mainta ining all QA/QC

procedures specified in the US EPA approved study protocol. As specified in the

US EPA approved study protocol, some bacte rial isolates were sent to IDEXX for

final confirmation using the VITEK bacterial identificat ion system. These

samples were coded at ASI and shipped to IDEXX with a well documented chain

of custody. Frozen archived samples for each bacterial isolate shipped to IDEXX

are being maintained at ASI.

Type of study:

The study was a side-by-side method comparison comparing each Colilert and

Colilert-18 against SM9222D, an EPA-approv ed reference method, to determine

whether the performance of both Colile rt and Colilert-18 are acceptable

compared to the reference method.

Study Schedule:

The data for this study was collected dur ing the months of October, November,

and December 2009. ASI was responsible for all aspects of the study and

reported the results to the Study Director on a weekly basis. All effluent samples

were tested at ASI within 6 hours of collection.

Matrix and spiking source:

The spiking source was primary sewage effluent from wastewater treatment

plants located in the New England area. These samples were collected and

tested according the US EPA’s Microbiological Alternative Test Procedure

Protocol for Drinking Water, Ambient Water, and Wastewater , Version 2004

(Appendix D, 1)

.

Preliminary testing:

Training was conducted at ASI during Oc tober 2009. Training consisted of

assessing ASI’s ability to accurately fo llow the US EPA approved study protocol

and to ensure that all processes and pr ocedures were well documented and

followed good QA/QC practices. Data collected by ASI during this training

demonstrated strong technical competence, exce llent documentation, and strong

quality standards.

Numbers and types of analyses:

Ten effluent samples were collected from ten different sewage treatment facilities

located in the New England area and used as fecal coliform sources for Colilert

and Colilert-18 vs. SM9222D comparability studies.

Sample collection, distribution, and storage:

Samples were collected in bottles containing preservatives as outlined in

Standard Methods for the Examination of Water and Wastewater , 20 th 

# Edition,

# ATP Case No. N09-004 Study Report

5 section 9060 A.2 (Appendix D, 2) . Samples were couriered to the laboratory in

coolers packed with ice or “blue ice”. As required in 40 CFR 136.3(e) Table II,

samples were analyzed within 6 hours of collection ( Appendix D , 2 ) . Chemical

characterization of the wastewat er samples was not required.

Data Reporting and Validation

Quality Assurance/Quality Control:

QA/QC procedures were followed as s pecified in the US EPA approved Study

Plan (Appendix B) and consistent with the guidelines of a certified laboratory.

Control cultures:

Each lot of medium was checked to ve rify the performance with known positive

and negative bacterial samples. Details of this testing can be found in the bound

data package provided by ASI (Appendix C)

.

Results

Fecal coliform recovery by eac h test method can be found in Tables 1 to 10

.

1 64.4 67.0 19.0

2 63.1 53.0 32.0

3 54.6 68.4 22.0

4 59.4 35.9 28.0

5 41.3 37.9 24.0

6 51.2 43.5 25.0

7 52.0 55.6 23.0

8 57.3 38.9 39.0

9 67.7 52.1 32.0

10 54.6 36.4 19.0

11 59.8 32.7 42.0

12 40.4 56.8 40.0

13 55.4 52.9 25.0

14 52.0 69.7 18.0

15 48.7 55.6 22.0

16 31.3 30.9 26.0

17 41.3 45.5 29.0

18 65.7 42.8 41.0

19 63.1 63.1 27.0

20 48.0 53.0 25.0 Table 1

Fecal Coliform Recovery from Bethel, ME Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

Average 53.57 49.59 27.90

# ATP Case No. N09-004 Study Report

6 1 83.6 38.4 17

2 62.7 47.1 61

3 69.1 24.9 49

4 71.7 39.9 44

5 61.3 51.2 55

6 69.7 28.5 41

7 59.4 59.1 41

8 62.4 29.2 29

9 60.5 34.1 38

10 52.1 61.7 29

11 55.6 52 30

12 93.3 36.8 46

13 57.3 55.6 57

14 63.1 47.1 36

15 55.6 39.3 31

16 58.6 34.1 31

17 61.3 48 50

18 83.6 59.4 33

19 68.3 42.6 43

20 65.7 36.4 30

Average 65.75 43.27 39.55 Table 2

Fecal Coliform Recovery from Burlington, VT Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

# ATP Case No. N09-004 Study Report

7 1 53.8 62.4 35

2 60.5 78.9 45

3 68.3 60.5 48

4 47.1 79.4 61

5 70.6 83.6 45

6 50.4 75.4 40

7 60.1 53.8 50

8 54.6 71.7 40

9 53.8 54.7 49

10 40.4 63.1 42

11 60.5 73.3 41

12 56.1 65 49

13 64.4 71.7 39

14 63.1 57.3 45

15 68.3 72.2 42

16 72.7 63.7 44

17 64.4 62.4 57

18 52 52.1 46

19 45.5 60.5 53

20 53 54.7 48

65.82 45.95 Average 57.98 Table 3

Fecal Coliform Recovery from W. Lebabnon, NH Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

1 44.8 78.4 31

2 63.7 80.9 45

3 70.6 63.8 53

4 48.7 56.1 42

5 49.6 71.2 43

6 41.3 62.4 42

7 59.1 66.3 49

8 46.4 74.3 45

9 32.7 69.7 33

10 74.9 52.8 44

11 54.6 58.3 34

12 52 51.2 38

13 53.8 69.1 34

14 45.7 64.4 40

15 52.1 66.3 57

16 38.9 40.4 45

17 50.4 43.5 51

18 41.9 44.1 49

19 55.7 52.1 47

20 40.4 66.3 48

61.58 43.50 Average 50.87 Table 4

Fecal Coliform Recovery from Keene, NH Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

# ATP Case No. N09-004 Study Report

8 1 45 54.6 53

2 47.9 45 43

3 36.8 52 54

4 47.9 34.1 51

5 49.5 38.4 41

6 32.3 45.5 48

7 45 41.9 51

8 33.1 36.9 47

9 36.4 42.6 47

10 44.1 33.6 59

11 44.8 38.4 37

12 35 45.7 41

13 48.7 40.4 44

14 49.5 45.7 16

15 45.7 31.7 33

16 48.7 41.9 43

17 26.9 59.8 44

18 31.3 52 42

19 45 60.1 47

20 46.4 49.5 26

Average 42.00 44.49 43.35 Table 5

Fecal Coliform Recovery from S. Burlington, VT Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)



# 1 44.8 65.1 29

2 71.7 65 37

3 68.3 65.7 42

4 65 66.3 32

5 48.7 63.7 30

6 32.8 63.1 42

7 52.9 73.3 39

8 54.7 77.6 33

9 55.6 65 41

10 47.9 69.7 36

11 56.5 55.7 29

12 46.4 69.7 9

13 53.8 57.3 29

14 47.1 83.6 37

15 52.1 63.1 29

16 62.4 78.9 36

17 40.4 69.1 28

18 43.5 55.6 26

19 55.6 70.3 19

20 41.9 76.6 34

67.72 31.85 Average 52.11 Table 6

Fecal Coliform Recovery from Essex Junction, VT Effluent



# Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

# ATP Case No. N09-004 Study Report

9 1 53.7 82 35

2 75.4 43.5 48

3 63.1 56.5 31

4 69.7 63.8 36

5 39.3 36.4 44

6 63.1 58.3 35

7 60.1 37.3 45

8 64.4 60.5 41

9 66.3 44.1 39

10 59.4 46.4 39

11 41.9 60.1 39

12 65 52.9 31

13 50.4 61.7 55

14 50.4 71.7 44

15 49.6 61.3 39

16 66.3 56.3 54

17 73.3 55.6 46

18 53.7 59.4 32

19 43.2 50.4 45

20 60.1 44.8 38

Average 58.4 55.2 40.8 Table 7

Fecal Coliform Recovery from Woodsville, NH Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

1 57.3 74.4 24

2 54.6 76.6 26

3 44.8 80.5 31

4 41.3 99 26

5 57.3 60.5 22

6 30.9 63.7 25

7 36.4 69.1 33

8 34.1 69.1 34

9 53.8 51.2 18

10 49.6 52.1 16

11 33.5 52.9 16

12 37.3 61.3 25

13 52 45.7 16

14 23.3 53 22

15 34.5 55.6 20

16 33.5 30.5 25

17 29.9 49.5 24

18 42.2 39.3 22

19 35 45.5 21

20 27.8 39.3 26

Average 40.5 58.4 23.6 Table 8

Fecal Coliform Recovery from Northhampton, MA Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

# ATP Case No. N09-004 Study Report

10 1 53.9 59.1 35

2 69.1 65.7 34

3 40.8 54.7 32

4 44.8 96 38

5 62.9 73.8 37

6 71.2 61.3 28

7 60.5 73.8 36

8 45.5 63.1 41

9 79.4 66.9 42

10 59.4 77.1 39

11 60.1 78 28

12 62.4 90.7 42

13 44.8 78 44

14 63.7 71.7 34

15 64.4 63.8 40

16 50.4 70.6 41

17 55.6 64.4 37

18 44.1 62.9 38

19 56.5 105 44

20 71.2 71.7 32

Average 58.0 72.4 37.1 Table 9

Fecal Coliform Recovery from Winthrop, MA Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)



# 1 33.1 96 40

2 47.1 98.7 42

3 41.6 98.8 43

4 33.5 93.3 52

5 37.9 77.6 39

6 28.8 49.6 41

7 36.8 81.6 44

8 4 1 8 5

. 5 5 1

9 37.9 81.6 55

10 44.8 90.7 54

11 34.1 60.1 26

12 31.3 77.1 42

13 44.8 69.7 54

14 41.3 86.2 58

15 28.8 105.4 34

16 33.5 64.4 50

17 42.8 74.9 35

18 25.6 63.8 41

19 42.6 88 51

20 36.9 76.6 40

Average 37.2 81.0 44.6 Table 10

Fecal Coliform Recovery from Littleton, NH Effluent

Replicate Colilert-18

(MPN/mL) Colilert

(MPN/mL) SM9222D

(cfu/mL)

# ATP Case No. N09-004 Study Report

11 Data Analysis and Discussion

Fecal coliform testing:

The ability to recover natural fecal colif orm populations was assessed for Colilert,

Colilert-18 and SM9222D usi ng primary effluent samp les collected from ten

different wastewater treat ment facilities located in the New England area ( Table

11 ) . Each test method was spiked wit h a primary effluent solution containing

approximately 20 to 50 fecal coliforms per 100mL in replicates of twenty per

effluent site. The test sa mples were incubated at 44.5 ± 0.2oC for the designated

period of time and analyzed to determi ne the presumptive fecal coliform

population recovered by each method. T he raw data for fecal coliform recovery

from this study can be found in Appendix C . The recovery data was analyzed to

assess the normality, precision, and re covery characteristics of each test

method.

# ATP Case No. N09-004 Study Report

Assessment of normality:

Fecal coliform recovery data for Colilert, Colilert-18, and SM9222D were

tested for normality with each effluent sample using the Kolmogorov-Smirnov test

( Table 12 ) . Analysis of these data indicated fe cal coliform recovery by all three

methods was consistent with a normal di stribution with p-values in excess of

0.366 for all samples with the exception of the Colilert-18 method, which had a pvalue of 0.119 with the South Burlington, Vermont effluen t. Even this p-value is

greater than the usual threshold of 0.10. These results confirm that the

assumption of normality has been met for a ll three test methods. According to

the ATP guidance document ( Appendix D , 1 ) quantitative methods that follow a

normal distribution need to be compar ed for variances between the effluent

samples to determine if the recovery variance is homogeneous across samples.

Assessment of precision:

Variance across ten effluent sites

The variance homogeneity in feca l coliform recovery across all matrices

for each test method was test ed using the Bartlett’s test ( Table 13 ) . For the

Colilert-18 method, the variances across th e ten sites were similar with a p-value

of 0.308. For Colilert and SM9222D the variances across the ten sites were not

homogeneous with p-values of 0.029 fo r Colilert and 0.006 for SM9222D.

Additional analysis using Levene’s test shows good internal consistency (p >

0.05) with the Colilert and Colilert-18 te sts but not with SM9222D. Overall, the

internal consistency was lowest with SM9222D, which seems to underscore the

difficulty in interpreting the results of this method with true environmental

samples. Microbiological methods that utilize morphological characteristics of

colonies as a means to detect a particular bacterial type are prone to this kind of

inconsistency given the subjectivity of this approach. The lack of internal

consistency required that the vari ances between methods be compared

12

# ATP Case No. N09-004 Study Report

separately for each effluent sample in stead of analyzing the pooled variances

from the ten effluent samples tested as per ATP guidelines.

Colilert-18 and Colilert vs. SM9222D

The variance equality in fecal coliform recovery between Colilert-18 and

SM9222D was tested using the F-test assu ming a normal distribution in recovery

with each effluent sample. The data presented in Table 14 show that fecal

coliform recovery by the Colilert-18 met hod was statistically equivalent (p > 0.05)

to SM9222D with 80% of the effluent sa mples tested. SM9222D showed higher

precision with two of the effluent samples (Northam pton and Winthrop,

Massachusetts).

The differences between varianc es between Colilert and SM9222D were

slightly more significant. The data presented in Table 15 show that six out of the

ten sites displayed similar variances (p>0 .05) between the two methods while the

remaining four sites (Northampton, Wint hrop, Littleton, and W oodsville) did not.

As noted above, SM9222D had the lowest overall precision across the ten

effluent samples tested by the three methods. This internal inconsistency

complicates the comparison of precision between the methods because such an

analysis assumes that SM9222D would pass this analysis. Given the relatively

poor precision of SM9222D in this study it’s difficult to interpret what a

comparison of its precision against t he Colilert and Colilert-18 methods really

means.

As previously mentioned, the re sults from SM9222D are difficult for an

analyst to read and interpret in a consist ent manner. Fecal coliform bacteria form

colonies on the m-FC medi um presenting colonies cont aining “various shades of

blue”, which can differ in intensity based on the metabolic state of the bacterium.

This detection strategy introduces consi derable subjectivity into reading the

results of this test based on the analysts’ interpretation of what constitutes a

shade of blue. Furthermore, non-feca l coliform bacteria grow on the m-FC

medium and produce colored co lonies ranging from gray to cream or even yellow

13

# ATP Case No. N09-004 Study Report

that can interfere with t he read. The presence of these competing bacterial

populations deprives fecal coliforms of important nutrients and depresses their

metabolic functions, thereby, decreasing the intensity of the blue color signal. It

is for this reason that SM9222D requi res all shades of blue be counted as

presumptive positive for fecal coliform det ection. In contrast, the Colilert and

Colilert-18 methods produce results that are unambiguous. With Colilert and

Colilert-18 the presence of fecal coliform bac teria is indicated by a distinct color

change in the wells of the Quanti-Tray device to yellow. This color change is

easy to read and interpret which is supported by the relatively high precision data

collected in this study, espec ially with the Colil ert-18 method. Finally, non-fecal

coliform bacteria are rarely able to grow and be detected by these test methods

so the intensity of the si gnal is more consistent than seen with SM9222D due to

the lack of competing flora.

Based on the above consideratio ns, Colilert and Colilert-18 methods

maintained superior internal consistency over SM9222D across all the effluents

samples tested. Inherent inconsis tencies seen with SM9222D preclude an

accurate assessment of the compar able variances between methods.

Assessment of recovery:

Colilert-18 vs. SM9222D

The ATP guidance document spec ifies that, when the assumption of

normality is not violated, the analyte re covery be first analyzed to determine if

significant interaction between method & ma trix occurred in the study. A Two

Way ANOVA analysis was used for this det ermination across all ten matrices,

which revealed that a significant intera ction (p = 0.00) wa s present between the

test methods and effluent samples (Appendix C) . As a result, the fecal coliform

recovery comparison was made

14

# ATP Case No. N09-004 Study Report

between methods with each independent effluent sample using a two sample ttest (a special case of ANOVA). T he results of this analysis can be found in

Table 16 and clearly show a bias in favor of Colilert-18 over SM9222D. Of the

ten effluents tested by this method, Coliler t-18 was found to recover a statistically

larger population of fecal coliform bacteri a (p < 0.05) than t he SM9222D in 80%

of the effluents tested. Colilert-18 was found to recover an equivalent amount of

fecal coliform bacteria (p = 0.623) from the South Burlington, Vermont effluent

and a statistically smaller population (p = 0.003) from the Littleton, New

Hampshire effluent.

Colilert vs. SM9222D

The results of this analysis ( Table 17 ) showed a clear bias in favor of the

Colilert method. Colilert was able to re cover a statistically larger population of

fecal coliform bacteria than SM9222D (p < 0. 05) in 80% of the effluents tested.

The remaining effluents (South Burlington & Burlington) yield ed equivalent fecal

coliform recovery (p > 0.05).

Both Colilert and Colilert-18 out performed SM9222D at recovering fecal

coliform bacteria from at least 80% of t he effluent samples tested. One factor

that may account for some of the differences seen in fecal coliform recovery

during this study is the discovery that SM9222D tended to underestimate the

concentration of fecal coliforms recove red on the m-FC medium. As will be

discussed in more detail below, SM9222D is prone to a much higher false

negative rate than the other tw o methods most likely due to inherent difficulties in

reading the results off the m-FC m edium as previously described.

Assessment of the false posit ive and false negative rates:

False positive

Presumptive positive responses from each of the te st methods were

harvested for further characterization fo llowing the methodology described in

15

# ATP Case No. N09-004 Study Report

Figure 1 . A minimum of 200 presumptive pos itive responses from each test

method were subjected to this confirma tion process to verify the accuracy of

each test. For the purposes of this st udy a fecal coliform was defined as: any

member of the Escherichia, Klebsie lla, Enterobacter, or Citrobacter genus

capable of growi ng at 44.5 ± 0.2 o C. This definition was applied equally to

Colilert, Colilert-18, and SM 9222D. The results of this study can be found in

Table 18 . All three test methods had low presumptive false positive rates

ranging from 0 to 1.5%. The majority of the presumptive positive samples were

confirmed as being true fecal coliform bacteria by the EC broth method.

However, less than 25% of the samples r equired further characterization by the

VITEK bacterial identification system because they failed to generate gas in the

EC medium when grown at 44.5 ± 0.2 o C. The identity of these anaerogenic fecal

coliform bacteria can be found in Tables 19

, 20 and 21 and the majority of the

time they were determined to be either Escherichia coli or Klebsiella

pneumoniae

.

Table 22 lists the identity of the confirm ed false positive isolates detected

by each test method. Two of the Colilert false posit ive samples were identified

as Raoultella planticola that formerly belonged to the Klebsiella genus. The

VITEK confidence level was lower than the 90% threshold specified by the study

protocol (was found to be 85%) so they were designated as false positive

responses. The same occurred with the one false positive sample detected by

SM9222D which technically fell below the th reshold of a fecal coliform as defined

by the study protocol.

False negative

Presumptive negative responses fr om each of the test methods were

harvested for further characterization following the methodology described in

Figure 2 . A minimum of 200 presumptive negat ive responses from each test

method were subjected to this confirma tion process to verify the accuracy of

each test to correctly identify true negativ e samples. The results of this study

16

# ATP Case No. N09-004 Study Report

can be found in Table 23 and show clear differences between the three test

methods. The confirmed false negative ra tes for the Colilert and Colilert-18

methods were found to be 9.0% and 7.0% , respectively. These values are

consistent with published reports and are com parable to each other in this study.

The VITEK identifications of these tr ue false negative samples can be found in

Tables 24

, 25 and 26 and are summarized in Table 27 by genus type. All the

false negative samples belonged to either the Escherichia or Klebsiella genus

with no specific pattern evident with regard to effluent type. Thr ee quarters of the

Colilert false negative responses belonged to the Escherichia genus and the

remaining quarter belonged to the Klebsie lla genus. The opposite distribution

was found with the Colilert-18 false negat ive samples. The confirmed false

negative rate of SM9222D was 21.5%, which far exceeded even the combined

rates of both the Colilert and Coliert -18 methods. As summarized in Table 27

,

the majority of the confirmed false negat ive responses for SM9222D belonged to

the genus Escherichia.

The relatively high false negat ive rate of SM9222D ma y explain the lower

recovery rate of fecal coliform bacteria observed with this method as compared

to Colilert and Colilert-18. More fe cal coliforms may have been present on the

m-FC medium that did not display the typi cal blue colony morphology specified in

the method so they were not included in the presumptive fecal coliform count.

The lack of a distinctive blue colony formation is possibly due to the presence of

non-target bacteria that co mpete for nutrients, and ther efore, interfere with the

growth and development of the fecal coliform colonies. It is also possible that the

strains that caused the false negative reac tion were weak to moderate lactose

fermenting strains were are not able to produce sufficient acid to react with the

m-FC indicator to produce the typical bl ue color. Weak lactose fermenting

strains were commonly encountered when confirming the presumptive positive

samples (see above). These strains failed to be confirmed with EC broth, which

depends upon lactose fermentation, and r equired subsequent confirmation by the

Vitek method to verify the fecal colif orm identity. The data presented in Table 18

showed that 33 out of the 200 (16.5%) presumptive positive samples obtained

17

# ATP Case No. N09-004 Study Report

from SM9222D were not able to ferment the lactose in EC broth. This value is

similar to the confirmed false negative rate for SM9222D and is consistent with

this explanation.

Study results indicate that Coli lert, Colilert-18, and SM9222D maintained a

very low and highly comparable false posit ive rate for the recovery of fecal

coliform bacteria from the effluent samp les tested. However, the inherent

variability of SM9222D coupled with its inabili ty to consistently detect low lactose

fermenting fecal coliforms resulted in a confirmed false negative rate which is

significantly higher than the more accura te Colilert and Co lilert-18 methods.

Conclusions:

Study results indicate that both th e Colilert and Colilert-18 methods have

met the US EPA acceptance criteria and have been shown to be equivalent or

better than SM9222D for detecting and quantify ing fecal coliforms in wastewater

samples. Both methods displayed a normal distribution for fecal coliform

recovery and maintained a statistically better internal consistency (i.e. precision)

across the ten effluent samples tested t han SM9222D. The lack of precision with

method SM9222D complicated the side by side comparisons of the relative

precision between methods therefore; no meaningful conclusion could be made.

Fecal coliform recovery by Colilert and Colilert-18 was statistically better

than SM9222D with at least 80% of the individual efflue nt samples when the site

specific recovery was compared. The hi gher overall recovery of fecal coliform

bacteria by Colilert and Colilert-18 was not due to the presence of false positive

responses as both methods exhibited fa lse positive rates of less than 2%.

However, the relatively high false negativ e rate of SM9222D (21.5%) seemed to

play a major role in the recovery differ ences. Detection of presumptive positive

fecal coliform bacteria by SM9222D relies on active lactose fermentation that

was found to be absent in 16.5% of the strains encounter ed in this study. These

anaerogenic fecal coliforms primarily belon ged to the Escherichia and Klebsiella

genus so they represented true fecal coliform bacteria and should have been

18

# ATP Case No. N09-004 Study Report

19 detected. Reading result s from SM9222D is also highly subjective and can be

complicated by the presence of competing bacterial populations that diminish the

intensity of the blue signal on m-FC medium . Evidence for this subjectivity was

found in the low precision data collected fr om SM9222D and from personal

communications from the anal ysts running the study.ATP Case No. N09-004Study ReportSTATECITYLOCATIONSHIPPING ADDRESSVERMONT ESSEX JUNCTIONESSEX JUNCTION WWTF2 Lincoln Street, Essex Junction, VT 05452VERMONT SOUTH BURLINGTON CITY OF SOUTH BURLINGTON WWTP1015 Airport Parkway, Burlington, VT 05403VERMONTBURLINGTONBURLINGTON MAIN WWTP53 Lavalley Lane, Burlington, VT 05401NEW HAMPSHIRE WOODSVILLEAOS WOODSVILLE WWTF9 Armory Lane, Woodsville, NH 03785NEW HAMPSHIRE WEST LEBANONWEST LEBANON WWTP130 South Main Street, West Lebanon, NH 03784NEW HAMPSHIRE LITTLETONAOS LITTLETON WWTP323 Meadow Street, Littleton, NH 03561NEW HAMPSHIRE KEENEKEENE WWTP420 Airport Road, Route 32, Keene, NH 03431MAINEBETHELBETHEL WWTP199 Main Street, Bethel, ME 04217MASSACHUSETTS NORTHAMPTONNORTHAMPTON WWTP33 Hockanum Road., Northampton, MA 01060MASSACHUSETTS WINTHROPMWRA (Deer Island Treatment Plant)Central Lab, 190 Tafts Avenue, Winthrop, MA 02152Table 11Primary Sewage Effluent Sample Sources20

# ATP Case No. N09-004 Study Report

Bethel

Maine 

# 37053 0.983 0.873 0.756

Northampton

Massachusetts 

# 37266 0.619 0.885 0.602

Winthrop

Massachusetts 

# 37265 0.88 0.538 0.995

Keene

New Hampshire 

# 37117 0.925 0.896 0.955

Littleton

New Hampshire 

# 37435 0.858 0.998 0.806

W. Lebanon

New Hampshire 

# 37188 0.989 0.832 0.961

Woodsville

New Hampshire 

# 37365 0.841 0.894 0.732

Burlington

Vermont 

# 37054 0.407 0.93 0.905

Essex Junction

Vermont 

# 37189 0.909 0.916 0.651

S. Burlington

Vermont 

# 37202 0.119 0.819 0.366 Table 12

Kolmogorov-Smirnov normality test p-values for data corresponding

to different sites and methods

Effluent site

tested P-value

(Colilert-18) P-value

(Colilert) P-value

(SM9222D) ASI effluent ID



# 0.125

0.042 Table 13

Test of variance equality for each method across the 10 sites

based on Bartlett’s test & Levene’s test p-values

Colilert

Method 9222D 0.308

0.029

0.006 Method P-value

(Barlett's test) P-value

(Levene's test)



# Colilert-18 0.619



# 21

# ATP Case No. N09-004 Study Report

Table 14

F-test p-values for comparison of variances between Colilert-18 and SM9222D

Effluent site

tested ASI effluent ID Std. Dev

(Colilert-18) Std. Dev

(SM9222D) P-value



# 0.279

Northhampton

Massachusetts 

# 37266 10.441 5.165 0.004 Bethel

Maine 

# 37053 9.662 7.504

0.001

Keene

New Hampshire 

# 37117 10.495 6.970 0.082 Winthrop

Massachusetts 

# 37265 10.602 4.756

0.178

W. Lebanon

New Hampshire 

# 37118 8.784 6.253 0.148 Littleton

New Hampshire 

# 37435 6.027 8.262

0.084

Burlington

Vermont 

# 37054 10.550 11.232 0.788 Woodsville

New Hampshire 

# 37365 10.290 6.849

0.391

S. Burlington

Vermont 

# 37202 7.136 9.842 0.170 Essex Junction

Vermont 

# 37189 9.735 7.969

Effluent site

tested ASI effluent ID Std. Dev

(Colilert) Std. Dev

(SM9222D)

Bethel

Maine 

# 37053 12.043 7.504

Northhampton

Massachusetts 

# 37266 16.331 5.165

Winthrop

Massachusetts 

# 37265 12.672 4.756

Keene

New Hampshire 

# 37117 11.494 6.970

Littleton

New Hampshire 

# 37435 14.506 8.262

W. Lebanon

New Hampshire 

# 37118 9.388 6.253

Woodsville

New Hampshire 

# 37365 11.178 6.849

Burlington

Vermont 

# 37054 10.895 11.232 0.00

0.00

S. Burlington

Vermont 

# 37202 8.201 9.842 Essex Junction

Vermont 

# 37189 7.566 7.969 0.90

0.82

0.43 Table 15

F-test p-values for comparison of variances between Colilert and SM9222D

0.06

0.02

0.09

0.04 P-value



# 0.05



# 22

# ATP Case No. N09-004 Study Report

C-18 SM9222D

* 

# cfu or MPN per 100mL sample 

# Table 16

Fecal coliform recovery comparison (Colilert-18 vs. SM9222D)

0.000

0.623 7.200

-0.500 0.000

0.000

0.000

0.013

0.003

0.000

0.000

0.000

37189 46.0 58.0

58.4 40.8

39.6 65.8

37202 9.380

6.470

8.060

2.610

-3.230

4.990

6.370

-7.600 53.6

Essex Junction

Vermont

S. Burlington

Vermont 

# 37053

37266

37265

37117

37435

37118

37365

37054 Littleton

New Hampshire

W. Lebanon

New Hampshire

Woodsville

New Hampshire

Burlington

Vermont

Bethel

Maine

Northhampton

Massachusetts

Winthrop

Massachusetts

Keene

New Hampshire 

# Effluent site

tested 

# ASI effluent ID T-value P-value Mean recovery *



# 27.9

40.5 23.6

37.1 58.0

50.9 43.5

44.6 37.2

52.1 31.9

42.0 43.4

CLT SM9222D

* 

# cfu or MPN per 100mL sample S. Burlington

Vermont 

# 37202 0.400 0.693 44.5 43.4 Essex Junction

Vermont 

# 37189 14.600 0.000 67.7 31.9 Burlington

Vermont 

# 37054 -1.060 0.295 43.3 39.6 Woodsville

New Hampshire 

# 37365 4.900 0.000 55.2 40.8 W. Lebanon

New Hampshire 

# 37118 7.880 0.000 65.8 46.0 Littleton

New Hampshire 

# 37435 9.750 0.000 81.0 44.6 Keene

New Hampshire 

# 37117 6.020 0.000 61.6 43.5 Winthrop

Massachusetts 

# 37265 11.670 0.000 72.4 37.1 Northhampton

Massachusetts 

# 37266 9.100 0.000 58.4 23.6 Bethel

Maine 

# 37053 6.830 0.000 49.6 27.9 Table 17

Fecal coliform recovery comparison (Colilert vs. SM9222D)

Effluent site

tested 

# ASI effluent ID T-value P-value Mean recovery *



# 23

# ATP Case No. N09-004 Study Report

a 

# Two of the wells confirmed by EC broth were initially negative for growth. Bacterial samples obtained from frozen archive

b 

# False positive rate = confirmed false positive wells or colonies/total number of wells or colonies collected for confirmation False positive rate b 

# 0.5% 

# Table 18

Confirmation of Presumptive Positive Responses by Each Test Method

200 166 33 1 12 8 0 0

5 1 4 0

0 Total (all sites) 

# 200 133

2 2 3 Colonies confirmed

(via. Vitek) 

# 15 1 19

S. Burlington, VT 

# 37202 

# 20 19

15 5 0 Burlington, VT 

# 37054 

# 20

Essex Junction, VT 

# 37189 

# 20 1 0

Woodsville, NH 

# 37365 

# 20 14 West Lebanon, NH 

# 37118 

# 20 18 Keene, NH 

# 37117 

# 20 16

Littleton, NH 

# 37435 

# 20 18 0 Northhampton, MA 

# 37266

Winthrop, MA 

# 37265 

# 20 17 20 17 False positive

colonies



# 0 0

3 0 Bethel, ME 

# 37053 

# 20 20 Effluent site

tested ASI effluent ID Colonies subjected

to confirmation Colonies confirmed

(via. EC broth) 

# 0

Fecal Coliform Positive Samples (SM9222D) 200 175 25 0

False positive rate b



# Total (all sites) S. Burlington, VT 

# 20 5 0

Essex Junction, VT 

# 37189 

# 20 16 4 0

Burlington, VT 

# 37054 

# 20 3 0 West Lebanon, NH 

# 37118

Woodsville, NH 

# 37365 

# 20 17 18 2 0

0 0

2 0 Littleton, NH 

# 37435 

# 20 20 Keene, NH 

# 37117 

# 20 18 2 0 Northhampton, MA 

# 37266

Winthrop, MA 

# 37265 

# 20 18 18 2 Wells confirmed

(via. Vitek) 

# False positive

wells



# 4 0

0 Bethel, ME 

# 37053 

# 20 16 Effluent site

tested ASI effluent ID Wells subjected

to confirmation Wells confirmed

(via. EC broth) 

# Fecal Coliform Positive Samples (Colilert Method)

Fecal Coliform Positive Samples (Colilert-18 Method) False positive rate b 

# 1.5% 5

7 1

0

45 3 1

0

0

0

0

0

14

13 1

0 4

9

6

3

3

4

1

3 17

16

19 a



# 17 15

11

13

17

20

20

20

20 20

20

20

20

37435 Northhampton, MA



# 37189

37202 S. Burlington, VT Essex Junction, VT

Woodsville, NH Winthrop, MA

Littleton, NH 

# 37053

37266

37265

37117

Total (all sites) 

# 0.0% 37118

37365

37054



# 20

20

20

20

37202 False positive

wells Effluent site

tested

Burlington, VT West Lebanon, NH Keene, NH Wells subjected

to confirmation ASI effluent ID Wells confirmed

(via. EC broth) Wells confirmed

(via. Vitek)

Bethel, ME



# 24

# ATP Case No. N09-004 Study Report

ASI

effluent ID Effluent site

tested ASI

well ID VITEK ID - Isolate name VITEK ID

confidence



# 307 Escherichia coli 99%

311 Escherichia coli 99%

312 Klebsiella pneumoniae ssp pneumoniae 97%

313 Escherichia coli 99%

622 Escherchia coli 99%

624 Enterobacter asburiae 96%

625 Klebsiella pneumoniae ssp pneumoniae 99%

627 Klebsiella pneumoniae ssp pneumoniae 96%

628 K. pneumoniae ssp pneumoniae & E. chloacae 99% & 98%

630 Klebsiella pneumoniae ssp pneumoniae 99%

636 Enterobacter cloacae 99%

637 K. pneumoniae ssp pneumoniae & E. chloacae 89% & 99%

639 Escherchia coli 99%

603 Raoultella planticola or Klebsiella pneumoniae N/A

611 Escherichia coli 99%

612 Klebsiella pneumoniae ssp pneumoniae 99%

614 K. pneumoniae ssp pneumoniae & E. chloacae 99%

615 Klebsiella pneumoniae ssp pneumoniae 99%

616 Klebsiella pneumoniae ssp pneumoniae 99%

617 Klebsiella pneumoniae ssp pneumoniae 95%

373 Klebsiella pneumoniae ssp pneumoniae 99%

374 Klebsiella pneumoniae ssp pneumoniae 97%

377 Escherichia coli 99%

766 Klebsiella pneumoniae or Klebsiella oxytoca N/A

767 Klebsiella pneumoniae ssp pneumoniae 91%

768 Klebsiella pneumoniae ssp pneumoniae 96%

382 Klebsiella pneumoniae ssp pneumoniae 94%

390 Klebsiella pneumoniae ssp pneumoniae 99%

394 Klebsiella pneumoniae ssp pneumoniae 99%

399 Klebsiella pneumoniae ssp pneumoniae 99%

37365 Woodsville

New Hampshire 711 Klebsiella pneumoniae ssp pneumoniae 96%

281 Klebsiella pneumoniae ssp pneumoniae 99%

291 Escherichia coli 99%

295 Escherichia coli 95%

484 Escherchia coli 99%

485 Enterbacter cloacae 99%

493 Klebsiella pneumoniae ssp pneumoniae 99%

497 K. pneumoniae ssp pneumoniae & E. coli 91% & 99%

499 Klebsiella pneumoniae ssp pneumoniae 99%

500 R. planticola or K. pneumoniae n/a

503 Escherichia coli 99%

507 Escherichia coli 99%

510 Escherichia coli 99%

515 Escherichia coli 99%

517 Klebsiella pneumoniae ssp pneumoniae 99%

518 Klebsiella pneumoniae ssp pneumoniae 99%

519 Escherichia coli 99%

= false positive because Vitek ID was less than 90% Bethel

Maine

37189

37202 37054 37118

Burlington

Vermont W. Lebanon

New Hampshire

Essex Junction

Vermont

S. Burlington

Vermont 

# Table 19

Confirmation of presumptive positive wells by Vitek (Colilert method)

Winthrop

Massachusetts Northampton

Massachusetts

Littleton

New Hampshire Keene

New Hampshire 37266 37053

37265

37435 37117



# 25

# ATP Case No. N09-004 Study Report

ASI

effluent ID Effluent site

tested ASI

well ID VITEK ID - Isolate name VITEK ID

confidence



# 242 Escherichia coli 99%

248 Escherichia coli 99%

256 Escherichia coli 99%

257 Escherichia coli 99%

587 Klebsiella pneumoniae ssp pneumoniae 99%

600 Klebsiella pneumoniae ssp pneumoniae 99%

575 Klebsiella pneumoniae ssp pneumoniae 99%

580 Escherichia coli 99%

331 Klebsiella pneumoniae ssp pneumoniae 92%

337 Escherichia coli 99%

345 Klebsiella pneumoniae ssp pneumoniae 99%

358 Klebsiella pneumoniae ssp pneumoniae 99%

683 Klebsiella pneumoniae ssp pneumoniae 99%

689 Escherichia coli 99%

693 Escherichia coli 98%

37054 Burlington

Vermont 270 Escherichia coli 99%

442 Klebsiella pneumoniae ssp pneumoniae 99%

446 Klebsiella pneumoniae ssp pneumoniae 99%

447 Klebsiella pneumoniae ssp pneumoniae 99%

450 Escherichia coli 99%

461 Escherichia coli 99%

464 Enterbacter sakazakii 95%

469 Escherichia coli 99%

472 Escherichia coli & Klebsiella pneumoniae 99%

478 Escherichia coli 99% 

# Table 20

Confirmation of presumptive positive wells by Vitek (Colilert-18 method)

37266 Northampton

Massachusetts

37365 Woodsville

New Hampshire 37053 Bethel

Maine

37202 S. Burlington

Vermont 37265 Winthrop

Massachusetts

37118 W. Lebanon

New Hampshire

37189 Essex Junction

Vermont 37117 Keene

New Hampshire



# 26

# ATP Case No. N09-004 Study Report

ASI

effluent ID Effluent site

tested ASI

colony ID VITEK ID - Isolate name VITEK ID

confidence



# 670 Klebsiella pneumoniae ssp pneumoniae 99%

675 Klebsiella pneumoniae ssp pneumoniae 99%

679 Klebsiella pneumoniae ssp pneumoniae 99%

648 Klebsiella pneumoniae ssp pneumoniae 99%

655 Klebsiella pneumoniae ssp pneumoniae 99%

657 Klebsiella pneumoniae ssp pneumoniae 91%

402 Escherichia coli 99%

408 Klebsiella pneumoniae ssp pneumoniae 99%

410 Klebsiella pneumoniae ssp pneumoniae 99%

411 Escherichia coli 99%

786 Escherichia coli 93%

787 Escherichia coli 99%

432 Klebsiella pneumoniae ssp pneumoniae 94%

436 Klebsiella pneumoniae ssp pneumoniae 99%

725 Klebsiella pneumoniae ssp ozaenae 85%

727 Escherichia coli 99%

729 Escherichia coli 94%

733 Enterobacter sakazakii 91%

735 Escherichia coli 95%

738 Escherichia coli 95%

37054 Burlington

Vermont 147 Klebsiella pneumoniae ssp pneumoniae 99%

521 Escherichia coli 99%

523 Escherichia coli 99%

525 Escherichia coli 99%

527 Escherichia coli 98%

531 Escherichia coli 99%

542 Klebsiella pneumoniae ssp pneumoniae 99%

543 Klebsiella pneumoniae ssp pneumoniae 99%

546 Klebsiella pneumoniae ssp pneumoniae 99%

547 Klebsiella pneumoniae ssp pneumoniae 99%

553 Klebsiella pneumoniae ssp pneumoniae 99%

555 Klebsiella pneumoniae ssp pneumoniae 99%

559 Klebsiella pneumoniae ssp pneumoniae 99%

560 Escherichia coli 99%

= false positive because Vitek ID was less than 90% 

# Table 21

Confirmation of presumptive positive colonies by Vitek (SM9222D)

Keene

New Hampshire Winthrop

Massachusetts

S. Burlington

Vermont Essex Junction

Vermont W. Lebanon

New Hampshire 37435 Littleton

New Hampshire

Woodsville

New Hampshire

Northampton

Massachusetts

37202 37265 37266

37365 37117

37118

37189



# 27

# ATP Case No. N09-004 Study Report

Woodsville, NH Klebsiella pneumoniae ssp ozaenae 85% 

# SM9222D 1 n/a n/a n/a 

# Colilert-18 0 85% n/a

Wintrop, MA Raoultella planticola or Klebsiella pneumoniae 85% 

# Colilert 3 Bethel, ME Non-viable bacterium

Essex Junction, VT Raoultella planticola or Klebsiella pneumoniae 

# Table 22

Fecal coliforms recovered from false positive wells or colonies

Test

method False positive

responses Effluent site Vitek ID Vitek confidence



# 28

# ATP Case No. N09-004 Study Report

a 

# False negative rate = confirmed false negative wells or colonies /total number of wells or colonies collected for confirmation 

# 20 17 

# Table 23

Confirmation of Presumptive Negative Responses by Each Test Method

Fecal Coliform Negative Samples (Colilert Method)

Effluent site

tested ASI effluent ID Wells subjected

to confirmation Wells confirmed

(via. EC broth) Wells confirmed

(via. Vitek) 

# False negative

wells



# 0 3

Northhampton, MA 

# 37266 

# 20 19 0 1 Bethel, ME 

# 37053

Winthrop, MA 

# 37265 

# 20 17

0 2

Keene, NH 

# 37117 

# 20 20 0 3

0 0

0 1 Littleton, NH 

# 37435

West Lebanon, NH 

# 37118 

# 20 19 20 18

Woodsville, NH 

# 37365 

# 20 18

0 0 Burlington, VT 

# 37054 

# 20 15 0 2

0 5

0 1 Essex Junction, VT 

# 37189

S. Burlington, VT 

# 37202 

# 20 19 20 20

Wells subjected

to confirmation Wells confirmed

(via. EC broth) 

# 18

False negative rate a 

# 9.0%

Fecal Coliform Negative Samples (Colilert-18 Method) Total (all sites) 

# 200 182 0

Wells confirmed

(via. Vitek) 

# False negative

wells

Bethel, ME 

# 37053 

# 20 20 0 0 Effluent site

tested ASI effluent ID

Northhampton, MA 

# 37266 

# 20 19

0 1

Winthrop, MA 

# 37265 

# 20 19 0 1

0 1

0 2 Keene, NH 

# 37117

Littleton, NH 

# 37435 

# 20 18 20 19

West Lebanon, NH 

# 37118 

# 20 15

0 1

Woodsville, NH 

# 37365 

# 20 18 0 5

0 2

0 0 Burlington, VT 

# 37054

Essex Junction, VT 

# 37189 

# 20 20 20 19

S. Burlington, VT 

# 37202 

# 20 19

False negative

colonies 

# 0 1

Total (all sites) 

# 200 186 0 14

20 9 False negative rate a 

# 7.0%

Fecal Coliform Negative Samples (SM9222D)

Effluent site

tested ASI effluent ID Colonies subjected

to confirmation Colonies confirmed

(via. EC broth) Colonies confirmed

(via. Vitek)



# 0 1 1

Northhampton, MA 

# 37266 

# 20 16 0 4 Bethel, ME 

# 37053

Winthrop, MA 

# 37265 

# 20 16

0 3

Keene, NH 

# 37117 

# 20 18 0 4

0 2

0 5 Littleton, NH 

# 37435

West Lebanon, NH 

# 37118 

# 20 15 36 33

Woodsville, NH 

# 37365 

# 17 11

0 3 Burlington, VT 

# 37054 

# 15 10 0 6

0 5

0 1 Essex Junction, VT 

# 37189

S. Burlington, VT 

# 37202 

# 20 19 17 14

44

False negative rate a 

# 21.5%

Total (all sites) 

# 205 161 0



# 29

# ATP Case No. N09-004 Study Report

ASI

effluent ID Effluent site

tested ASI

well ID VITEK ID - Isolate name VITEK ID

confidence



# 306 Escherichia coli 99%

311 Escherichia coli 99%

319 Escherichia coli 98%

37266 Northampton

Massachusetts 622 Escherchia coli 94%

606 Escherichia coli 96%

607 Escherichia coli 88%

612 Escherichia coli 99%

767 Escherichia coli 99%

780 Escherichia coli 99%

37118 W. Lebanon

New Hampshire 389 Escherichia coli 96%

722 Klebsiella pneumoniae ssp pneumoniae 99%

740 Escherichia coli 99%

283 Escherichia coli 99%

287 Klebsiella pneumoniae ssp pneumoniae 99%

288 Escherichia coli & Enterobacter aerogenes 94% & 98%

289 Klebsiella pneumoniae ssp pneumoniae 95%

297 Klebsiella pneumoniae ssp pneumoniae 99%

37202 S. Burlington

Vermont 516 Escherichia coli 96% 

# Table 24

Confirmation of presumptive negative wells by Vitek (Colilert method)

37365 Woodsville

New Hampshire 37435 Littleton

New Hampshire 37265 Winthrop

Massachusetts 37053 Bethel

Maine

37054 Burlington

Vermont



# 30

# ATP Case No. N09-004 Study Report

ASI

effluent ID Effluent site

tested ASI

well ID VITEK ID - Isolate name VITEK ID

confidence



# 37266 Northampton

Massachusetts 597 Escherichia coli 99%

37265 Winthrop

Massachusetts 568 Klebsiella pneumoniae ssp pneumoniae 99%

37117 Keene

New Hampshire 335 Klebsiella pneumoniae ssp pneumoniae 99%

743 Escherichia coli 98%

750 Klebsiella pneumoniae ssp pneumoniae 99%

344 Klebsiella pneumoniae ssp pneumoniae 99%

347 Escherichia coli & Klebsiella pneumoniae 99% & 95%

348 Klebsiella pneumoniae ssp pneumoniae 99%

351 Klebsiella pneumoniae ssp pneumoniae 99%

359 Klebsiella pneumoniae ssp pneumoniae 99%

681 Klebsiella pneumoniae ssp pneumoniae 99%

698 Klebsiella pneumoniae ssp pneumoniae 99%

37054 Burlington

Vermont 268 Escherichia coli 99%

37202 S. Burlington

Vermont 461 Klebsiella pneumoniae ssp pneumoniae 94% 37435 Littleton

New Hampshire 

# Table 25

Confirmation of presumptive negative wells by Vitek (Colilert-18 method)

37365 Woodsville

New Hampshire 37118 W. Lebanon

New Hampshire



# 31

# ATP Case No. N09-004 Study Report

ASI

effluent ID Effluent site

tested ASI

colony ID VITEK ID - Isolate name VITEK ID

confidence



# 123 Escherichia coli 99%

124 Escherichia coli 99%

125 Escherichia coli 99%

126 Escherichia coli 99%

130 Escherichia coli 99%

132 Escherichia coli 99%

134 Escherichia coli 99%

135 Escherichia coli & Klebsiella pneumoniae 99%

136 Escherichia coli 99%

137 Escherichia coli 99%

138 Escherichia coli 99%

661 Klebsiella pneumoniae ssp pneumoniae 94%

669 Escherichia coli 99%

673 Escherichia coli 99%

674 Klebsiella pneumoniae ssp pneumoniae 99%

642 Klebsiella pneumoniae ssp pneumoniae 95%

643 Klebsiella pneumoniae ssp pneumoniae 99%

651 Klebsiella pneumoniae ssp pneumoniae 94%

655 Klebsiella pneumoniae ssp pneumoniae 95%

416 Escherichia coli & Klebsiella pneumoniae 99%

418 Escherichia coli 99%

790 Escherichia coli 99%

793 Escherichia coli 99%

795 Escherichia coli 99%

426 Escherichia coli 99%

430 Escherichia coli 99%

432 Escherichia coli 99%

435 Escherichia coli 99%

438 Escherichia coli 99%

721 Escherichia coli 99%

723 Klebsiella pneumoniae ssp pneumoniae 99%

726 Enterbacter sakazakii 99%

730 Escherichia coli 99%

733 Escherichia coli 99%

734 Escherichia coli 99%

142 Klebsiella pneumoniae ssp pneumoniae 94%

144 Klebsiella pneumoniae ssp pneumoniae 98%

153 Escherichia coli 98%

156 Escherichia coli 99%

158 Escherichia coli 99%

521 Escherichia coli 99%

529 Klebsiella pneumoniae ssp pneumoniae 99%

531 Escherichia coli 99%

37202 S. Burlington

Vermont 548 Escherichia coli 99% 

# Table 26

Confirmation of presumptive negative colonies by Vitek (SM9222D)

37189 Essex Junction

Vermont 37265 Winthrop

Massachusetts

37118 W. Lebanon

New Hampshire 37117

37435 Littleton

New Hampshire Keene

New Hampshire

37054 Burlington

Vermont 37053 Bethel

Maine

37266 Northampton

Massachusetts

37365 Woodsville

New Hampshire



# 32

# ATP Case No. N09-004 Study Report

75%

2%

0% 

# SM9222D Klebsiella 23% 

# 44 Escherichia

Enterobacter

Citrobacter 27%

0%

0% 

# Colilert-18 Klebsiella 71% 

# 14 Escherichia

Enterobacter

Citrobacter Enterobacter

Citrobacter 78%

0%

0% Klebsiella 22% 4

0 

# Colilert 18 Test

method False negative

responses Genera Frequency Occurrence

14 Escherichia

0 

# Table 27

Fecal coliforms recovered from false negative wells or colonies

0

33

10

1 0

4

10

0



# 33

# ATP Case No. N09-004 Study Report



# Figure 1

Confirmation Procedure for Positive Response Samples

Colilert-18/Q-tray 2000

Colilert/Q-tray 2000

Method 9222D

Select 1 “positive response”

from each replicate for

verification

Frozen (-70oC)

archived source LTB incubated at

35±0.5oC for 48±3 hrs

EC Broth incubated at

44.5±0.2oC for 24±2 hrs

Gas & growth No Gas & growth

True Positive

Nutrient Agar slant

incubated at 35±

0.5oC for 18 to 24 hrs

VITEK

Fecal Coliform non-fecal coliform

True Positive Possible

False Positive McConkey agar

plate incubated at 35±

0.5oC for 18 to 24 hrs 

# 10μL aliquot

or colony 800μL from Colilert &

Colilert-18 samples

800μL from

9222D samples

10μL aliquot

10μL aliquot

10μL aliquot

Frozen (-70oC)

archived source

False Positive Has archived

source been

screened?

(2 cycles)



# YES (after 2 rounds of testing) NOAll typical and atypical coliform colony types



# <90% confidence =90% confidence EMB Agar

incubated at 35±

0.5oC for 18 to 24 hrs



# Typical fecal coliform colony types



# 34

# ATP Case No. N09-004 Study Report



# Figure 2

Confirmation Procedure for Negative Response Samples

Colilert-18/Q-tray 2000

Colilert/Q-tray 2000

Method 9222D

Select 1 “negative response”

from each replicate for

verification

Frozen (-70oC)

archived source LTB incubated at

35±0.5oC for 48±3 hrs

EC Broth incubated at

44.5±0.2oC for 24±2 hrs

Gas & growth No Gas & growth

False Negative

Nutrient Agar slant

incubated at 35±

0.5oC for 18 to 24 hrs

VITEK ID 

# 10μL aliquot

or colony 800μL from Colilert &

Colilert-18 samples

800μL from

9222D samples

10μL aliquot

True Negative 

# Turbid

Growth?

YES NO

McConkey Agar

incubated at 35±

0.5oC for 18 to 24 hrs 

# 10μL aliquot

Typical fecal coliform colony types



# 35

# ATP Case No. N09-004 Study Report



# APPENDIX A(1) Method

IDEXX Colilert ® - 

# 18 TEST METHOD FOR THE DETECTION OF

FECAL COLIFORMS IN WASTEWATER

36

# ATP Case No. N09-004 Study Report



# APPENDIX A(1) Method

1.0 Scope and Application

1.1 This method is intended for use in the detection and enumeration of fecal

coliform bacteria in wastewater samp les without additional confirmation

steps; any positive response (yellow well s) is an indication of the presence

of fecal coliform bacteria

1.2 The minimum, non-zero number of bac terial counts detectable with this

method is a function of any diluti on scheme used when processing the

sample

1.3 The Colilert-18 ® 

# method, when used in conjunction with Quanti-Tray ®



# or

Quanti-Tray ® 

# /2000, is a quantification met hod capable of detecting up to

2419MPN/100mL fecal coliforms without a dilution (see package insert)

(1)

1.4 Since there can be wide range of fecal coliform populations in wastewater

samples, dilutions can be used wit h this method for detecting and

enumerating the actual concentration



# 2.0 Summary of Method

2.1 The method is based on De fined Substrate Technology ® 

# , when reagent is

added to the wastewater sample and incubated, it can detect fecal

coliform bacteria at 1 CFU/100mL wit hin 18 hours with as many as 2

million heterotrophic bacte ria present per 100 mL

2.2 The Colilert-18 tests are manufactured in snap packs; one snap pack

contains enough powder to perform a si ngle test for the presence of fecal

coliform bacteria in a 100mL wastewater sample. Contents of one snap

pack is added to a vessel containi ng 100mL of a wastewater sample

followed by mixing (shaking) to dissolve the powder. The sample plus

medium is then transferred to a sterile Quanti-Tray device then placed into

the Quanti-Tray sealer, which disperse s and seals the sample into a large

number of incubating wells. The Q uanti-Tray is then incubated at 44.5

±0.2°C for 18 hours. After incubati on, the number of yellow wells are

counted and compared against a mo st probable number (MPN) chart

(provided with the Quanti-Trays) to arri ve at the final number of fecal

coliform bacteria detected in the original wastewater sample. If a dilution

is used, the MPN/100mL is corrected based on the dilution. If needed, the

yellow wells can be compared against a co lor comparator-. to verify the

result. Any yellow color present in the wells which meets or exceeds the

color of the comparator is considered to be positive for the presence of

fecal coliform bacteria. The presenc e of this yellow color under these

incubation conditions is specific for t he detection of fecal coliform bacteria.

No additional confirmati on steps are required.



# 3.0 Definitions

3.1 In this method, fecal coliform bacteri a are those bacteria which produce a

yellow color after incubation at 44.5°C± 0.2oC for 18 hours, specifically

37

# ATP Case No. N09-004 Study Report



# APPENDIX A(1) Method

those bacteria in Escherichia, Klebs iella, Enterobacter, or Citrobacter

genus capable of growing at 44.5 ± 0.2oC

3.2 Fecal coliform detection is based on the presence of the enzyme ß-Dgalactosidase which is known to be present in fecal coliform bacteria

3.3 For the detection of ß-D-galactosi dase, Colilert-18 utilizes the

chromogenic nutrient indicator ort ho-nitrophenyl-ß-D- galactopyranoside

(ONPG), which produces a distinct yellow color when hydrolyzed by ß-Dgalactosidase



# 4.0 Interferences

4.1 Heterotrophic bacteria greater than 2,000,000/100ml can yield a positive

response for the presence of fecal coliform bacteria (1)

4.2 Samples containing humic material may have an innate color and a

control blank of the same sample may be required for comparison to the

inoculated sample (1)



# 5.0 Safety

5.1 The analyst/technician must know and observe the normal safety

procedures required in a microbiology laboratory preparing, using, and

disposing of samples, reagents and materials, and while operating

sterilizing equipment.

5.2 Any waste containing potentially viable microorganisms should be

decontaminated by following specific State and/or Federal guidelines



# 6.0 Equipment and Supplies

6.1 Pipettes, sterile, T.D. bacteriologic al or Mohr, glass or plastic of

appropriate volume

6.2 Quanti-Tray or Quanti-Tray 2000 tr ays (IDEXX part numbers WQT100 and

WQT-2K, respectively)

6.3 Quanti-Tray Sealer (IDEXX part number WQTS2X-115)

6.4 Sterile vessels, glass or plastic, 120-250ml volume

6.5 Incubator/Water Bath maintained at 44.5°C±0.2°C

6.6 EZ QC (quality control bacterial strains) (IDEXX part number WEZQ)



# 7.0 Reagents

7.1 Sterile deionized water or distilled water; water conforming to specification

D1193, “Reagent water conforming to Type II, Annual Book of ASTM

Standards” (2) ; autoclave at 121°C (15 -lb pressure) for 15 minutes or

sterile filter using a 0.22 micron filter into a sterile container

7.2 Colilert-18 snap packs, store at 2°C- 25°C away from light. The expiration

date is indicated on the package ( 15 months from the date of

manufacture). (1) ; IDEXX part numbers WP020I -18 or WP200I-18)

7.3 Antifoam drops or vessels (IDEXX part number WAFDB)

7.4 Sodium thiosulfate

38

# ATP Case No. N09-004 Study Report



# APPENDIX A(1) Method

8.0 Sample Collection, Preservation and Storage

8.1 Sampling procedures are described in detail in the US EPA microbiology

methods manual, Section II, A (18.3) (3) Samples were collected in bottles

containing preservatives as outlined in Standard Methods for the

Examination of Water and Wastewater , 20 th 

# Edition, se ction 9060 A.2. (4)

8.2 Storage Temperature and H andling Conditions: Ice or refrigerate

bacteriological samples at a temp erature less than 10°C ( 2°C-10°C)

during transit to the laboratory (3) ; use insulated containers to assure

proper maintenance of stor age temperature. Ensure that sample vessels

are not totally immersed in water during transit

8.3 Holding Time Limitations: Examine samples as soon as possible after

collection. For wastewater comp liance, do not exceed 6 hours holding

time and process samples within 2 hours

8.4 Chlorinated wastewater samples s hould be neutralized at time of

collection with sodium thiosulfate



# 9.0 Quality Control

9.1 Quality control should be conducted on each lot of Colilert-18 or more

often as regulations requires. Inoculat e 100 ml of sterile water with either

EZ QC or ATCC strains as listed below . Follow the procedure in section

11.0 and then refer to this char t for results interpretation:

EZ QC Bacteria Expected Result

EZ QC Escherichia coli Yellow color production

EZ QC Pseudomonas

aeruginosa No color change



# 10.0 Calibration and Standardization

10.1 Check the temperature in the incubator daily to ensure it is within the

stated limits

10.2 Check thermometers at least annually against NIST certified thermometer

or one that meets t he requirements of NI ST Monograph SP 250-23



# 11.0 Procedure

11.1 Carefully separate one blister pack fr om the strip taking care not to

accidentally open the adjacent pack

11.2 Ensure the powder is in t he bottom of the blister pack

11.3 Hold the blister pack face down (paper side up) at the top and towards the

bottom and snap back at the scoreline forming a “V”, with the opening

facing into the open vessel.

11.4 Allow the powder to fall into the water sample contained in the sterile

vessel

39

# ATP Case No. N09-004 Study Report



# APPENDIX A(1) Method

11.5 If a dilution is required, use sterile dei onized or distilled water, not buffered

water for making the dilutions. Alwa ys add Colilert-18 to the final 100ml

diluted sample only

11.6 If foaming occurs add Antifoam solu tion to the sample containing the

reagent (1) ; alternatively, allow the vessel to sit for several minutes prior to

adding it to a Quanti-Tray

11.7 Aseptically cap and seal the vessel

11.8 Mix / shake gently to dissolve

11.9 Remove either a sterile Quanti-Tray or Quanti-Tray/2 000 tray from the

plastic bag (tear open the plastic bag at the bottom which has a green line

around the bag) and squeeze the tray at the top to open it; pour the

sample reagent mixture from the vessel into the tray avoiding contact with

the foil tab

11.10 Seal the tray with t he Quanti-Tray sealer

11.11 Incubate at 44.5° ± 0.2°C

11.12 Read the results at 18 hours. If needed, compare the yello w color of the

wells with either a Quanti-Tra y or Quanti-Tray 2000 comparator

11.13 If no yellow color is observed in the wells, the sample is absence of fecal

coliforms and recorded as <1/100mL

11.14 If the sample has a yellow color equal to or greater than the comparator,

the presence of fecal coliforms is confirmed

11.15 If the sample is yellow, but not dist inguishable from the color comparator,

it may be incubated up to an additi onal 4 hours (but no more than 22

hours total). If the color is then equal to or greater than the comparator,

the sample is positive for fecal colif orms; if it does not change, it is

negative



# 12.0 Data Analysis and Calculations

12.1 Count the number of positive wells.

12.2 Refer to the Quanti-Tray MPN t able provided by IDEXX to determine the

most Probable Number (MPN) for total coliforms (yellow wells) (1)

12.3 Correct the MPN value for any dilution made.

12.4 Report results as MPN/100mL for fecal coliforms.



# 13.0 Method Performance

13.1 The Colilert-18 

# method is a superior and more reliable test for the detection

and enumeration of fecal coliform bacteria from wastewater than the

SM9222D method as described in: ATP Study Report for the Proposed US

EPA Approval of Detection and Enumer ation of Fecal Coliforms in US

Wastewater Samples 

# (5)



# 14.0 Pollution Prevention

14.1 The solutions and reagents used in this method pose no threat to the

environment when recycled and managed properly.

40

# ATP Case No. N09-004 Study Report



# APPENDIX A(1) Method

14.2 Solutions and reagents should be pr epared in volumes consistent with

laboratory use to minimize the volume of expired materials to be disposed.



# 15.0 Waste Management

15.1 It is the laboratory’s respons ibility to comply with a ll federal, state and local

regulations governing waste managem ent, particularly the biohazard and

hazardous identification ru les and land disposal regulations. Compliance

with all sewage discharge permits and regulations is also required.

15.2 Samples, reference materials and equi pment known or suspected to have

viable bacteria attached or contained must be sterilized prior to disposal.



# 16.0 References

1. Draft Colilert-18 Package Insert and Q uanti-Tray and Quanti-Tray/2000

Package Inserts from IDEXX

2. Annual Book of ASTM Standards, Vol. 11.01, American Society for

Testing Materials, Philadelphia, PA 19103, (2005)

3. Bordner, R., J.A. Wint er and P.V. Scarpino (eds.) Microbiological

Methods for Monitoring the Environment, Water and Wastes, EPA-600/8-

78-017. Office of Research and Deve lopment, USEPA. (December 1978)

4. Clesceri, L.S., A.E. Greenberg, A. D. Eaton (eds.). 1998 Standard

Methods for the examination of Water and Waster Water, 20 th 

# Edition,

American Public Health Asso ciation, Washington, DC.

5. ATP Study Report for the Proposed US EP A Approval of Detection and

Enumeration of Fecal Coliforms in US Wastewater Samples, January

2010

41

# ATP Case No. N09-004 Study Report



# APPENDIX A(2) Method

IDEXX Colilert ® 

# TEST METHOD FOR THE DETECTION

OF FECAL COLIFORMS IN WASTEWATER

42

# ATP Case No. N09-004 Study Report



# APPENDIX A(2) Method

1.0 Scope and Application

1.1 This method is intended for use in the detection and enumeration of fecal

coliform bacteria in wastewater samp les without additional confirmation

steps; any positive response (yellow well s) is an indication of the presence

of fecal coliform bacteria

1.5 The minimum, non-zero number of bac terial counts detectable with this

method is a function of any diluti on scheme used when processing the

sample

1.6 The Colilert ® 

# method, when used in c onjunction with Quanti-Tray ® 

# or

Quanti-Tray ® 

# /2000, is a quantification method capable of detecting up to

2419MPN/100mL fecal coliforms without a dilution (see package insert)

(1)

1.7 Since there can be wide range of fecal coliform populations in wastewater

samples, dilutions can be used wit h this method for detecting and

enumerating the actual concentration



# 2.0 Summary of Method

2.1 The method is based on De fined Substrate Technology ® 

# ; when the

reagent is added to the wastewater sa mple and incubated, it can detect

fecal coliform bacteria at 1 CFU/100m L within 24 hours with as many as 2

million heterotrophic bacte ria present per 100 mL

2.2 The Colilert tests are manufac tured in snap packs; one snap pack

contains enough powder to perform one test for the presence of fecal

coliform bacteria in 100mL of wastewat er. Add the contents of one snap

pack to a vessel containing 100mL of a wastewater sample followed by

mixing (shaking) to dissolve the powder. This is transferred to a sterile

Quanti-Tray device and is then place into the Quanti-Tray sealer which

dispenses and seals the reagent into a large number of incubating wells.

The Quanti-Tray is then incubated at 44.5 ±0.2°C for 24 hours. After

incubation, the number of yellow wells are c ounted and compared against

a most probable number (MPN) chart (provided with the Quanti-Trays) to

arrive at the final number of fecal colif orm bacteria detected in the original

wastewater sample. If a dilution is used, the MPN/100mL is corrected

based on the dilution. If needed, the yellow wells can be compared against

a color comparator-. to verify the re sult. Any yellow color present in the

wells which meets or exceeds the color of the comparator is considered to

be positive for the presence of fecal coliform bacteria. The presence of

this yellow color under these incubation conditions is specific for the

detection of fecal coliform bacteria. No additional confirmation steps are

required.

43

# ATP Case No. N09-004 Study Report



# APPENDIX A(2) Method

3.0 Definitions

3.1 In this method, fecal coliform bacteri a are those bacteria which produce a

yellow color after incubation at 44.5° C±0.2°C for 24 hours, specifically

those bacteria in Escherichia, Klebs iella, Enterobacter, or Citrobacter

genus capable of growing at 44.5 ± 0.2oC

3.2 Fecal coliform detection is based on the presence of the enzyme ß-Dgalactosidase which is known to be present in fecal coliform bacteria

3.3 For the detection of ß-D-galactosidase, Colilert utilizes the chromogenic

nutrient indicator ortho-nitrophenyl-ß- D-galactopyranoside (ONPG), which

produces a distinct yellow color w hen hydrolyzed by ß-D-galactosidase



# 4.0 Interferences

4.1 Heterotrophic bacteria greater than 2,000,000/100mL can yield a positive

response for the presence of fecal coliform bacteria (1)

4.2 Samples containing humic material may have an innate color and a

control blank of the same sample may be required for comparison to the

inoculated sample (1)



# 5.0 Safety

5.1 The analyst/technician must know and observe the normal safety

procedures required in a microbiology laboratory preparing, using, and

disposing of samples, reagents and materials, and while operating

sterilizing equipment

5.2 Any waste containing potentially viable microorganisms should be

decontaminated by following specific State and/or Federal guidelines



# 6.0 Equipment and Supplies

6.1 Pipettes, sterile, T.D. bacteriologic al or Mohr, glass or plastic of

appropriate volume

6.2 Quanti-Tray or Quanti-Tray 2000 tr ays (IDEXX part numbers WQT100 and

WQT-2K, respectively)

6.3 Quanti-Tray Sealer (IDEXX part number WQTS2X-115)

6.4 Sterile vessels, glass or plastic, 120-250mL volume

6.5 Incubator/Water Bath maintained at 44.5°C±0.2°C

6.6 EZ QC (quality control bacterial strains) (IDEXX part number WEZQ)



# 7.0 Reagents

7.1 Sterile deionized water or distilled water; water conforming to specification

D1193, “Reagent water conforming to Type II, Annual Book of ASTM

Standards” (2); autoclave at 121°C (15 -lb pressure) for 15 minutes or

sterile filter using a 0.22 micron filter into a sterile container

7.2 Colilert snap packs, store at 2°C-25°C away fr om light. The expiration

date is indicated on the package ( 15 months from the date of

manufacture). (1) (IDEXX part numbers WP020I or WP200I)

44

# ATP Case No. N09-004 Study Report



# APPENDIX A(2) Method

7.3 Sodium thiosulfate



# 8.0 Sample Collection, Preservation and Storage

8.1 Sampling procedures are described in detail in the US EPA microbiology

methods manual, Section II, A (18.3) (3) Samples were collected in bottles

containing preservatives as outlined in Standard Methods for the

Examination of Water and Wastewater , 20 th 

# Edition, se ction 9060 A.2. (4)

. 8.2 Storage Temperature and H andling Conditions: Ice or refrigerate

bacteriological samples at a temp erature less than 10°C ( 2°C-10°C)

during transit to the laboratory (3) . Use insulated containers to assure

proper maintenance of stor age temperature. Ensure that sample vessels

are not totally immersed in water during transit.

8.3 Holding Time Limitations: Examine samples as soon as possible after

collection. For wastewater comp liance, do not exceed 6 hours holding

time and process samples within 2 hours.

8.4 Chlorinated wastewater samples should be neutralized at time of

collection with sodium thiosulfate



# 9.0 Quality Control

9.1 Quality control should be conducted on each lot of Colilert or more often

as regulations requires. Inoculate 100 mL of ster ile water with either EZ

QC or ATCC strains as listed below. Follow the procedure in section 11.0

and then refer to this chart for results interpretation:

EZ QC Bacteria Expected Result

EZ QC Escherichia coli Yellow color production

EZ QC Pseudomonas

aeruginosa No color change



# 11.0 Calibration and Standardization

10.1 Check the temperature in the incubator daily to ensure it is within the

stated limits.

10.2 Check thermometers at least annually against NIST certified thermometer

or one that meets the requirements of NIST Monograph SP 250-23.



# 11.0 Procedure

11.1 Carefully separate one blister pack from the strip taking care not to

accidentally open the adjacent pack

11.2 Ensure the powder is in t he bottom of the blister pack

11.3 Hold the blister pack face down (paper side up) at the top and towards the

bottom and snap back at the scoreline forming a “V”, with the opening

facing into the open vessel

45

# ATP Case No. N09-004 Study Report



# APPENDIX A(2) Method

11.4 Allow the powder to fall into the water sample contained in the sterile

vessel

11.5 If a dilution is required, use sterile deionized or distilled water, not buffered

water for making the dilutions. Al ways add Colilert to the final 100mL

diluted sample only.

11.6 Aseptically cap and seal the vessel

11.7 Mix / shake gently to dissolve

11.8 Remove either a sterile Quanti-Tray or Quanti-Tray/2000 tray from the

plastic bag (tear open the plastic bag at the bottom which has a green line

around the bag) and squeeze the tray at the top to open it; pour the

sample reagent mixture from the vessel into the tray avoiding contact with

the foil tab

11.9 Seal the tray with t he Quanti-Tray sealer

11.10 Incubate at 44.5° ± 0.2°C

11.11 Read the results at 24 hours. If needed, compare the yello w color of the

wells with either a Quanti-Tra y or Quanti-Tray /2000 comparator

11.12 If no yellow color is observed in the wells, the sample is absence of fecal

coliforms and recorded as <1/100mL

11.13 If the sample has a yellow color equal to or greater than the comparator,

the presence of fecal coliforms is confirmed

11.14 If the sample is yellow, but not dist inguishable form the color comparator,

it may be incubated up to an additi onal 4 hours (but no more than 28

hours total); if the color is then equal to or greater than the comparator,

the sample is positive for fecal colif orms; if it does not change, it is

negative



# 12.0 Data Analysis and Calculations

12.1 Count the number of positive wells

12.2 Refer to the Quanti-Tray MPN t able provided by IDEXX to determine the

most Probable Number (MPN) for total coliforms (yellow wells) (1)

12.3 Correct the MPN value for any dilution made

12.4 Report results as MPN/100mL for fecal coliforms



# 13.0 Method Performance

13.1 The Colilert method is a superior and more reliable test for the detection

and enumeration of fecal coliform bacteria from wastewater than the

SM9222D method as described in: ATP Study Report for the Proposed

US EPA Approval of Detection and En umeration of Fecal Coliforms in US

Wastewater Samples (5)



# 14.0 Pollution Prevention

14.1 The solutions and reagents used in this method pose no threat to the

environment when recycled and managed properly

46

# ATP Case No. N09-004 Study Report



# APPENDIX A(2) Method

14.2 Solutions and reagents should be pr epared in volumes consistent with

laboratory use to minimize the volume of expired materials to be disposed



# 15.0 Waste Management

15.1 It is the laboratory’s respons ibility to comply with all federal, state and local

regulations governing waste managem ent, particularly the biohazard and

hazardous identification ru les and land disposal regulations. Compliance

with all sewage discharge permits and regulations is also required

15.2 Samples, reference materials and equi pment known or suspected to have

viable bacteria attached or contained must be sterilized prior to disposal



# 16.0 References

1. Draft Colilert Package Insert and Q uanti-Tray and Quanti-Tray/2000

package inserts from IDEXX

2. Annual Book of ASTM Standards, Vol. 11.01, American Society for

Testing Materials, Philadelphia, PA 19103, (2005)

3. Bordner, R., J.A. Wint er and P.V. Scarpino (eds.) Microbiological

Methods for Monitoring the Environment, Water and Wastes, EPA-600/8-

78-017. Office of Research and Deve lopment, USEPA. (December 1978)

4. Clesceri, L.S., A.E. Greenberg, A. D. Eaton (eds.). 1998 Standard

Methods for the examination of Water and Waster Water, 20 th 

# Edition,

American Public Health Asso ciation, Washington, DC.

5. ATP Study Report fo r the Proposed US EP A Approval of Detection and

Enumeration of Fecal Coliforms in US Wastewater Samples. January

2010

47

# ATP Case No. N09-004 Study Report

48 

# Appendix D

Supporting References

1. EPA Microbiological Alternate Test Pr ocedure (ATP) Protocol for Drinking

Water, Ambient Water, and Wastewater Monitoring Methods , Guidance; EPA

821-B-03-004; Environmental Protec tion Agency, Office of Water,

Washington, DC 20460 2004

2. Code of Federal Regulations 40 CFR 136.3

3. APHA. 1998. Standard Methods for the Ex amination of Water and

Wastewater. 20 th 

# Edition. American Public H ealth Association. 1015 15 th



# Street, NW, Washington, DC 20005.

# UNITED STATES ENVIRONMENTAL PROTECTION AGENCY



# WASHINGTON, D.C. 20460



# Patsy Root

Regula t ory Affairs Specia l ist

IDEXX Labora t ories, Inc.

I IDEXX Drive

Westbrook ME 04092

Dear Ms

. Root: 

# AUG 

# 0 3 2009

The Engineering and Analytica l Support Branch ( E ASB) has completed a

technical review of the method valida t ion study p l an submitted in support of t he

fo ll owing alternate test procedure (ATP) application. OFFICE OF



# WATER



# ATP Case No's. Analyte Study Plan

(wastewater)

N09-0004 Fecal coliform Revis i on 1.0, 7 / 21/2009

Addendum 1.0, 7 / 31/2009

We ar e pleased to inform you that in the judgement of our technical staff the proposed

study plan is adequate for the A TP program. We look forward to reviewing the comp l ete

data package when i t is available

. P l ease feel free to contact us i f you have any

additional questions re g arding these applications

.

Robin K. 

# Osh i ro

, Ph

. D

Microbiolog i s t

Engineering and Analy s is Division

EASB Sincere l y,

# ;;i_{l**~**

Lemue l Walker

A TP Coordinator

Engineering and Ana l ys i s Divisio n

EASB

lntemet Address (URL) • http : //www

. epa

. gov Recycled/Recyclable • Printed with Vegetable Oi l Based I nks on 100% Postconsumer

, Process Chlorine Free Recycled Paper

# EPA Case # N09-004



# Study Plan

Study Protocol for the Proposed US EPA Approval of

IDEXX’s Colilert ® 

# -18 and Colilert ® 

# for the Detection and

Enumeration of Fecal Coliforms in Wastewater Samples

July 21, 2009

IDEXX Laboratories, Inc.

One Idexx Drive

Westbrook, Maine 04092

1

# EPA Case # N09-004

Method Description

1.0 Scope and Application

1.1 Colilert ® 

# -18 and Colilert ® 

# are Defined Substrate ® 

# methods for the

detection of fecal coliforms in wastewater samples

1.2 The sample type for the study is primary effluent, from wastewater

treatment plants

1.3 Colilert-18 and Colilert used in co mbination with ID EXX’s Quanti-Tray ®



# and/or Quanti-Tray ® 

# /2000 MPN methods allow for detecting and

enumerating fecal coliforms at 18 hours (hrs) for Co lilert-18 and/or 24 hrs

for Colilert after incubating at 44.5 ± 0.2 ° C without additiona l confirmation

or verification steps

1.4 The limit of detection of Colilert-18 and Colilert is 1 colony-forming unit

(cfu) of fecal coliforms per 100 mL of sample

2.0 Summary of Method

2.1 Colilert ® 

# -18 and Colilert ® 

# used in combination with IDEXX’s Quanti-Tray ®



# and/or Quanti-Tray ® 

# /2000 methods detect and enumerate fecal coliforms

in wastewater samples

2.2 Colilert-18 and Colilert ar e based on the detection of β -D-galactosidase,

which are indicative of the feca l coliform group when the samples are

incubated at 44.5 ± 0.2 ° C; as the target microbes grow, they will hydrolyze

chromogenic nutrient indicator specific for β -D-galactosidase;

antimicrobial agents are pres ent in the test reagent to suppress the growth

of most non-target microorganisms

2.3 Colilert-18 and Colilert both cont ain the chromogenic substrates, ortho

nitrophenyl- β -D-galactopyranoside (ONPG); upon hydrolysis by the fecal

coliform bacteria at 44.5 ± 0.2 ° C, the ONPG in Colilert-18 and Colilert/or

release a chromogenic compound, ortho -nitrophenol (ONP), which

changes the color of the test fr om colorless to yellow

2.4 A blister pack of the reagent (Colilert- 18 or Colilert) powder is added to a

sterile test vessel containing 100 mL of a wastewater sample; after the

powder is added, the vessel is sh aken to dissolve the powder; the

dissolved sample/medium mixture is added to a Quanti-Tray or QuantiTray/2000 and sealed with a Quanti-Tray sealer; the tray is then placed

2

# EPA Case # N09-004

into an incubator at t he specific temperature and time required for the

specific test as indicated in 1.3 above

3.0 Definitions

3.1 ONPG (ortho-nitrophenyl- β -D-galactopyranoside) – A nutrient indicator

containing a food source attached to a chromogen. The fecal coliform

bacteria contain an enzyme, β -D-galactosidase that cleaves the food

source, galactopyranoside and releases the chromogen, ONP, to yield a

yellow color

3.2 Hydrolysis – Biochemical reaction catalyzed by a bacterial enzyme

resulting in the decomposition of a substrate by the addition of a water

molecule

3.3 Fecal Coliforms – A bacterial subset belonging to the Enterobacteriaciae

family and defined biochemically by the presence of the enzyme β -Dgalactosidase and will grow at 44.5 ± 0.2 ° C in both Colilert and Colilert-18

reagent

3.4 Quanti-Tray ® 

# – An automated MPN method that is used with IDEXX’s

Colilert and Colilert-18 te sts for the enumeration of s pecific target bacteria

with a counting range up to 200 cfu/100 mL

3.5 Quanti-Tray ® 

# /2000 – An automated MPN me thod that is used with

IDEXX’s Colilert and Colilert-18 tests fo r the enumeration of specific target

bacteria with a counting range up to 2419 cfu/100 mL

4.0 Interferences

4.1 There are no known interferences contai ned in wastewater which interfere

with the performance of these test methods

5.0 Safety

5.1 The analyst must practice normal safety procedures for working in a

microbiological laboratory and handling pot entially bio-hazardous cultures

5.2 Solid and liquid waste containing viable microorganisms should be

decontaminated by following specific State and/or Federal guidelines

5.3 Colilert ® 

# -18 and Colilert ® 

# are not listed as a suspected carcinogen. Refer

to the Material Safety Data Sheet (MSDS) provided with the product for

specific information relating to the potential hazards of this product by

accidental ingestion or exposure

3

# EPA Case # N09-004

6.0 Quality Assurance/Quality Control

6.1 Quality Assurance: The laboratorie s participating in the study have

comprehensive quality assurance program s in place. The laboratories are

certified for microbiological analysis of water and wastewater by both their

respective state laboratory certificati on programs and by the United States

Environmental Protection Agency

6.2 Quality Control: All QC procedures should be performed as specified in

the methods. The study report will incl ude sufficient raw data so that an

independent reviewer can verify each determination and calculation

performed by the laboratory or the st udy coordinator. This verification

consists of tracing all st eps of the method to the fi nal result reported. The

raw data are, generally, method-spec ific and may include but are not

limited to the following:

6.2.1 Sample numbers or other ident ifiers used by the laboratory

6.2.2 Sample collection dates and times

6.2.3 Verification that method-specified QC procedures were met for test

samples and all associated QC samples

6.2.4 Analysis dates and times for all steps in the method

6.2.5 Sample volume

6.2.6 Any measurements of ancillary para meters (i.e., temperature, pH,

turbidity, percent solids, etc.)

6.2.7 Results for all intermediate steps in the method

6.2.8 Preliminary data steps to determine the final result

6.2.9 Final result

6.2.10 If appropriate, quantitation reports , data system outputs, and other

data to link the raw data to the results reported

6.2.11 Laboratory bench sheets and copies of all pertinent logbook pages

for all sample preparation and cle anup steps, and for all other parts

of the determination

6.2.12 Temperature logs for water bath, incubators, refri gerators, etc.

6.2.13 Media preparation information

6.2.14 If appropriate, direct instrument re adouts and other data to support

the final results

7.0 Equipment and Suppli es: (not exhaustive)

7.1 Equipment:

7.1.1 Air incubators capable of mainta ining temperatures at 35±0.5 ° C

4

# EPA Case # N09-004

7.1.2 Air incubators capable of maintain ing temperatures at 44.5±0.2 ° C

(see Appendix A, incubat or specification)

7.1.3 Water bath maintained at 50 ° C for tempering agar

7.1.4 Water bath capable of maintainin g temperatures at 44.5±0.2oC

7.1.5 IDEXX Quanti-Tray ® 

# Sealer

7.1.6 VITEK ® 

# automated bacteriological ident ification system, to be used

to resolve discrepant results obtai ned from confirmation procedures

for both positive and negative responses

7.1.7 All equipment required for Standard Method 9222D

7.2 Glassware and supplies – all glassware and plastic vessels used for

determining the presence of the bacteria species should be handled

according to recommended guidelines an d in keeping with good laboratory

practices; all glassware and contai ners used in this study should be

washed with a suitable laboratory det ergent, rinsed thoroughly with tap

water followed by rinsing with reagent water to remove residual detergent,

and sterilized by autoclaving at 121 ° C for 15 minutes prior to use

7.2.1 IDEXX sterile plastic 120 mL collection vessels w/o sodium

thiosulfate

7.2.2 Colilert ® 

# -18 and Colilert ® 

# tests pre-packaged in disposable blister

packs

7.2.3 Disposable sterile pipettes (1, 5, 10, 25, and 50mL)

7.2.4 Calibrated Pipetteman and sterile disposable tips

7.2.5 IDEXX Quanti-Tray ® 

# and/or Quanti-Tray ® 

# /2000 devices

7.2.6 Antifoam for use with Colilert-18

7.2.7 All materials required for the Standard Method 9222D

8.0 Comparability & Specificity Study for the Detection and Enumeration of

Fecal Coliforms in Wastewater Samples

8.1 The study will be performed following t he general procedure described in

the Appendix of the EPA Microbiological Alternate Test Procedure (ATP)

Protocol for Drinking Water, Ambient Water, and Wastewater Monitoring

Methods. Guidance – April 2004

8.2 The study will be conducted at Analytic al Services, Inc. (130 Allen Brook

Lane, Williston, Vermont), a ce rtified reference laboratory

8.3 Comparability Experimental Set-Up Outline (Figure 1)

8.3.1 At least ten primary sewage efflue nt wastewater samples known to

contain fecal coliforms will be collected from mu ltiple States within

and surrounding the greater New England region. Samples will be

collected in bottles containing pres ervatives as outlined in the 20th

5

# EPA Case # N09-004

Ed. of Standard Method , 9060 A.2. Samples will be couriered to

the laboratory in coolers packed with ice or “blue ice”. Samples will

be analyzed within 6 hours of collect ion. Chemical characterization

of the wastewater samples is not required and will not be performed

8.3.2 The fecal coliform concentration in each wastewater sample will be

estimated using the Colilert ® 

# -18/Quanti-Tray ® 

# /2000,

Colilert ® 

# /Quanti-Tray ® 

# /2000 methods, and Standard Method

9222D (m-FC medium)

8.3.3 The wastewater sample will be seria lly diluted in sterile de-ionized

water to obtain multiple (3 to 5) dilution sets of samples for the

comparability testing in order to generate at least one set of 20

replicates with approx imately 20-50 cfu of fecal coliforms per 100

mL of test sample

8.3.4 One hundred milliliter aliq uots of the diluted wastewater samples

will be tested with the Colilert- 18/Quanti-Tray/2000 method (in

replicates of 20 for each dilu tion) following manufacturer’s

instructions

8.3.5 One hundred milliliter aliq uots of the diluted wastewater samples

will be tested with Colile rt/Quanti-Tray/2000 method (in replicates of

20 for each dilution) following ma nufacturer’s instructions

8.3.6 One hundred milliliter aliq uots of the diluted wastewater samples

will be tested with the m-FC medium (in replicates of 20 for each

dilution) following Standard Method 9222D instruction

8.3.7 Colilert and Colilert-18 will be incubated in a dry 44.5±0.2oC

incubator that has been documented to hold the 44.5oC

temperature within 0.2oC throughout the chamber, over a 24 hour

period as described in Standard Methods for the Examination of

Water and Wastewater , 21 st 

# Ed, 2005, page 9- 66 (Appendix A); the

m-FC standard method will be incubated in a 44.5±0.2oC water

bath in accordance with the standard method procedure

8.3.8 The Colilert/Quanti-Tray/2000 me thod will be incubated for 24

hours

8.3.9 The Colilert-18/Quanti-Tray/2000 method will be incubated for 18

hours

8.3.10 The Standard Method 9222D samples will be incubated for 24±2

hours

6

# EPA Case # N09-004

8.3.11 After incubation, each test method will be read to determine the

number of presumptive pos itive responses (yellow wells in the case

of Colilert & Colile rt-18 and blue colonies in the case of Standard

Method 9222D

8.3.12 One of the dilution sets in which Standard Method 9222D yields

approximately 20 to 50 presumpt ive positive colonies will be

retained for further analysis of the false positive and false negative

error for fecal coliform detection

8.3.13 An assessment of normality (verif ication of normal distribution of

results) will be done for all test me thods with statistical tests as

specified by the EPA Microbiological Alternate Test Procedure

(ATP) Protocol for Drinking Water, Ambient Water, and Wastewater

Monitoring Methods, Guidance (page 8.5)

8.3.14 An assessment of precision will be done for all test methods as

specified by the EPA Microbiological Alternate Test Procedure

(ATP) Protocol for Drinking Water, Ambient Water, and Wastewater

Monitoring Methods, Guidance (page 8.5 – 8.6)

8.3.15 Fecal coliform recovery by each method will be evaluated as

specified by the EPA Microbiological Alternate Test Procedure

(ATP) Protocol for Drinking Water, Ambient Water, and Wastewater

Monitoring Methods, Guidance (page 8.8 – 8.9)

8.3.16 Presumptive positive and presum ptive negative responses from

each of the test methods at the chosen dilution will be subjected to

a further verification study to arrive at the relative false positive and

false negative rates

8.4 Determination of the False Positive Error for Fecal Coliform

Detection (Figure 2)

8.4.1 Identify one presumptive positive (ye llow) well from each Colilert

and Colilert-18/Quanti-Tray/2000 replicate for further verification

(20 samples from each method per dilution) using the following

method

8.4.1.1 Use an alcohol wipe to disinfect the paper/foil backing of

the “presumptive positive” well identified on the QuantiTray/2000 device

8.4.1.2 Use a sterile pipette tip to poke through the back of the

well, extract 800 μ L of sample and dispense into a cryo7

# EPA Case # N09-004

vial containing 200 μ L of sterile glycerol. Label and store

the vial in a -70oC freezer as a sample archive

8.4.1.3 Using a new, sterile pipe tte tip, extract another 10 μ L

aliquot and transfer into Lauryl Tryptose Broth for fecal

coliform confirmation following Standard Method

9221B.1b

8.4.1.4 The Lauryl Tryptose Broth is incubated at 35±0.5 ° C for

48±3 hours (as per Standard Method 9221B.1)

8.4.1.5 Transfer a 10 μ L aliquot into EC Broth (as per Standard

Method 9221E) and incubate in a 44.5±0.2oC water bath

for 24±2 hours

8.4.1.6 The presence of gas and turbid growth in the EC Broth is

a confirmed positive response for a fecal coliform

8.4.1.7 The absence of gas is considered a negative response for

the presence of a fecal coliform

8.4.1.8 Streak 10 μL aliquots from all non-gassing samples onto

EMB agar plates and incubate at 35±0.5oC for 18 to 24

hours

8.4.1.9 Streak typical fecal coliform colony types from EMB plate

onto Nutrient Agar slants/plat es and incubate at 35±0.5oC

for 18 to 24 hours

8.4.1.10 Confirm bacterial identification from each Nutrient agar

slants/plates using a VITEK Bacterial ID system

8.4.1.11 A VITEK identificati on of greater than 90% confidence is

considered a confirmed positive response for the

detection of a fecal coliform

8.4.2 Identify one presumptive positiv e (blue) colony from each Standard

Method 9222D replicate for further verification (20 samples from

each method per dilution) using the following method

8.4.2.1 Inoculate these colonies into Lauryl Tryptose Broth for

fecal coliform confirmation following Standard Method

9221B.1b

8.4.2.2 The Lauryl Tryptose Broth is incubated at 35±0.5 ° C for

48±3 hours (as per Standard Method 9221B.1)

8

# EPA Case # N09-004

8.4.2.3 Transfer 800 μ L of sample into a cryo-vial containing 200

μ L of sterile glycerol. Label and store the vial in a -70oC

freezer as a sample archive

8.4.2.4 Transfer another 10 μ L aliquot into EC Broth (as per

Standard Method 9221E) and incubate in a 44.5±0.2oC

water bath for 24±2 hours

8.4.2.5 The presence of gas and turbid growth in the EC Broth is

a confirmed positive response for a fecal coliform

8.4.2.6 The absence of gas is considered a negative response for

the presence of a fecal coliform

8.4.2.7 Streak 10 μL aliquots from all non-gassing samples onto

EMB agar plates and incubate at 35±0.5oC for 18 to 24

hours

8.4.2.8 Streak typical fecal coliform colony types from EMB plate

onto Nutrient Agar slants/pl ates and incubate at 35±0.5oC

for 18 to 24 hours

8.4.2.9 Confirm bacterial identification from each Nutrient agar

slants/plates using a VITEK Bacterial ID system

8.4.2.10 A VITEK identificati on of greater than 90% confidence is

considered a confirmed positive response for the

detection of a fecal coliform

8.4.3 If the presence of a fecal coliform is confirmed in EC Broth or by the

VITEK ID system, the result is interpreted as a True Positive

Response for the presence of a fecal coliform by the test method

(i.e. Colilert, Colilert-18, or Standard Method 9222D).

8.4.4 If a negative response for the pres ence of a fecal coliform is

obtained, test the frozen archived sample using the following

method

8.4.4.1 Streak 10μL of frozen sample onto a MacConkey agar

plate and incubate for 18 to 24 hours in a 35±0.5 oC

incubator

8.4.4.2 Inoculate all “typical” and “atypical” coliform colony types

into EC Broth and incubate in a 44.5±0.2oC water bath for

24±2 hours

9

# EPA Case # N09-004

8.4.4.3 The presence of gas and turbid growth in the EC Broth is

considered a confirmed positive response for the

presence of a fecal coliform

8.4.4.4 If none of the turbid tubes contain gas streak 10 μ L

aliquots onto EMB agar plates and incubate at 35±0.5oC

for 18 to 24 hours

8.4.4.5 Streak all colony types from EMB plates onto nutrient agar

plates/slants and incubate at 35±0.5oC for 18 to 24 hours

8.4.4.6 Confirm bacterial identificat ion from each Nutrient agar

slants/plates using a VITEK Bacterial ID system

8.4.4.7 A VITEK identificati on of greater than 90% confidence is

considered a confirmed positive response for the

detection of a fecal coliform bacterium

8.4.5 If the presence of a fecal coliform is confirmed in EC Broth or by the

VITEK method, the result is interpreted as a True Positive

Response for the presence of a fecal coliform by the test method

(i.e. Colilert, Colilert-18, or Standard Method 9222D)

8.4.6 Failure to confirm the presence of a fecal coliform by 2 cycles of the

method described in section 8.4.3 is interpreted as a False

Positive Response for fecal coliform by the test method

8.4.7 This assay will be repeated until the validity of at least 200 positive

responses have been tested from a ll three fecal coliform methods

8.5 Determination of the False Negative Error for Fecal Coliform

Detection (Figure 3)

8.5.1 Identify one presumptive negative (col orless) well from each Colilert

and Colilert-18/Quanti-Tray/2000 replicate for further verification

(20 samples from each method per dilution) using the following

method

8.5.1.1 Use an alcohol wipe to disinfect the paper/foil backing of

the “presumptive negative” well identified on the QuantiTray/2000 device

8.5.1.2 Use a sterile pipette tip to poke through the back of the

well, extract 800 μ L of sample and dispense into a cryo10

# EPA Case # N09-004

vial containing 200 μ L of sterile glycerol. Label and store

the vial in a -70oC freezer as a sample archive

8.5.1.3 Using a new, sterile pipe tte tip, extract another 10 μ L

aliquot and transfer into Lauryl Tryptose Broth for fecal

coliform confirmation following Standard Method

9221B.1b

8.5.1.4 The Lauryl Tryptose Broth is incubated at 35±0.5 ° C for

48±3 hours (as per Standard Method 9221B.1)

8.5.1.5 The absence of turbid growth is considered a confirmed

negative response for the pres ence of a fecal coliform

8.5.1.6 If turbid growth is present, transfer a 10 μ L aliquot into EC

Broth (as per Standard Method 9221E) and incubate in a

44.5±0.2oC water bath for 24±2 hours

8.5.1.7 The presence of gas and turbid growth in the EC Broth is

considered a confirmed positive response for a fecal

coliform

8.5.1.8 The absence of gas is considered a confirmed negative

response for the presence of a fecal coliform

8.5.2 Identify one presumptive negative (grey or cream colored) colony

from each Standard Method 9222D replicate for further verification

(20 samples from each method per dilution) using the following

method

8.5.2.1 Inoculate these colonies into Lauryl Tryptose Broth for

fecal coliform confirmation following Standard Methods

9221B.1b

8.5.2.2 The Lauryl Tryptose Broth is incubated at 35±0.5 ° C for

48±3 hours (as per Standard Method 9221B.1)

8.5.2.3 The absence of turbid growth is considered a confirmed

negative response for the pres ence of a fecal coliform

8.5.2.4 If turbid growth is present, transfer 800 μ L of sample into

a cryo-vial containing 200 μ L of sterile glycerol. Label and

store the vial in a -70oC freezer as a sample archive

11

# EPA Case # N09-004

8.5.2.5 Transfer another 10 μ L aliquot into EC Broth (as per

Standard Method 9221E) and incubate in a 44.5±0.2oC

water bath for 24±2 hours

8.5.2.6 The presence of gas and turbid growth in the EC Broth is

a confirmed positive response for a fecal coliform

8.5.2.7 The absence of gas is considered a confirmed negative

response for the presence of a fecal coliform

8.5.3 If a confirmed negative respons e for the presence of a fecal

coliform is obtained, the result is interpreted as a True Negative

Response for the presence of a fecal coliform by the test method

8.5.4 If a confirmed positive response for the presence of a fecal coliform

is obtained, the result is interpreted as a False Negative

Response for the presence of a fecal coliform by the test method

8.5.5 Streak all confirmed positive sa mples from the EC Broth onto

MacConkey agar plates and incubate at 35±0.5oC for 18 to 24

hours

8.5.6 Transfer all typical fecal coliform colony types onto Nutrient Agar

slants/plates and incubate at 35±0.5oC for 18 to 24 hours

8.5.7 Confirm the bacterial identification from all the Nutrient agar slants

using a VITEK Bacterial ID system

8.5.8 This assay will be repeated until t he validity of at least 200 negative

responses have been tested for all three test methods

8.6 False Positive and False Negative Rate Calculation

8.6.1 False Positive Rate = FP/(TP) X 100%

FP = False Positives, which are the number of initial positive

responses by the method which are not verified by the confirmation

procedure

TP = Total Positives, which are the number of total positive

responses (both confirmed and non-c onfirmed positive samples) by

the test method

8.6.2 False Negative Rate = FN/(TN) X 100%

12

# EPA Case # N09-004

FN = False Negatives, which are the number of initial negative

responses by the method which are verified by the confirmation

procedures to contain true positive samples

TN = Total Negatives, which are the number of total negative

responses (both confirmed and non- confirmed negative samples)

by the test method

13

# EPA Case # N09-004

Figure 1. Overall Expe rimental Flow Chart

Sewage Effluent Samples

Analyzed within 30 hrs of collection and

4 hrs of arrival at the laboratories

Prepare multiple dilution samples in sterile

de-ionized water



# Test Methods

Colilert-18/Quanti-Tray 2000 and

Colilert/Quanti-Tray 2000

3-5 Dilutions and each dilution with 20

replicates for each method 

# Method 9222D (m-FC)

3-5 Dilutions and each

dilution with 20 replicates

Add Colilert-18 and/or Colilert reagent

to 100mL diluted wastewater sample,

allow the reagent to dissolve and pour

the reagent/sample mixture in a

Quanti-Tray 2000 subsequently sealed

by the Quanti-Tray Sealer Filter 100mL aliquot of

diluted wastewater sample

through a membrane and

place the membrane onto

the mFC medium.

Incubate at 44.5 ± 0.5oC for 18 hrs for

the Colilert-18/Q-Tray 2000 method &

24 hrs Colilert/Q-Tray method Incubate the mFC medium at

44.5 ± 0.2 ° C for 24 +/- 2 hrs

Count the number of wells with positive

response (yellow color) for Colilert-18

and Colilert) for fecal coliforms.

Convert to MPN/100mL using IDEXX

MPN Table Count the number of blue

colonies as presumptive

fecal coliforms



# 14

# EPA Case # N09-004

Figure 2. Confirmation Procedur e for Positive Response Samples

Colilert-18/Q -tray 2000

Colilert/Q -tray 2000

M ethod 9222D

Select 1 “positive response”

from each replicate for

verification

Frozen (-70oC)

archived source LTB incubated at

35±0.5oC for 48±3 hrs

E C B roth incubated at

44.5±0.2oC for 24±2 hrs

G as & growth N o G as & growth

True Positive

N utrient A gar slant

incubated at 35±

0.5oC for 18 to 24 hrs

VITEK

Fecal C oliform non-fecal coliform

True Positive P ossible

False Positive McConkey agar

plate incubated at 35±

0.5oC for 18 to 24 hrs 

# 10μL aliquot

or colony 800μL from C olilert &

C olilert-18 sam ples

800μL from

9222D sam ples

10μL aliquot

10μL aliquot

10μL aliquot

Frozen (-70oC)

archived source

False Positive H as archived

source been

screened?

(2 cycles)



# YE S (after 2 rounds of testing) N OAll typical and atypical coliform colony types



# <90% confidence =90% confidence EMB Agar

incubated at 35±

0.5oC for 18 to 24 hrs



# T ypical fecal coliform colony types



# 15

# EPA Case # N09-004

16 Figure 3. Confirmation Procedur e for Negative Response Samples

Colilert-18/Q-tray 2000

Colilert/Q-tray 2000

Method 9222D

Select 1 “negative response”

from each replicate for

verification

Frozen (-70oC)

archived source LTB incubated at

35±0.5oC for 48±3 hrs

EC Broth incubated at

44.5±0.2oC for 24±2 hrs

Gas & growth No Gas & growth

False Negative

Nutrient Agar slant

incubated at 35±

0.5oC for 18 to 24 hrs

VITEK ID 

# 10μL aliquot

or colony 800μL from Colilert &

Colilert-18 samples

800μL from

9222D samples

10μL aliquot

True Negative 

# Turbid

Growth?

YES NO

McConkey Agar

incubated at 35±

0.5oC for 18 to 24 hrs 

# 10μL aliquot

Typical fecal coliform colony types

# Addendum to ATP Case 

# # N09-004

IDEXX Laboratories, Inc.

July 31, 2009

EPA Question #1 -"definition of fecal coliform as defined by your method, to wit,

Colilert/18 Quanti-Tray/2000 positive result when run not at the current

instruction specification of 35C but at 44.5C (with the plus minus degrees)"

IDEXX response

- A fecal coliform is defined as a subset of the total coliform

population capable of being detected by the Colilert/Colilert-18 technology after

incubation at 44.5 ±0.2C as indicated by the presence of a yellow well in the

Quanti-Tray/Quanti-Tray 2000 test device.

EPA Question #2- "verification via VITEK, where an identification of greater

than 90% confidence of the following genera xxx will be considered a confirmed

response for the detection of fecal coliform"

IDEXX response: "verification via VITEK, where an identification of greater than

90% confidence, with either SM 92220 or IDEXX method, of the following

genera: Escherichia, Citrobacter, Klebsiella, and Enterobacter, will be considered

a confirmed response for the detection of fecal coliform"

# P~t5Y)R~,

# 0;;~~------

Regulatory Affairs Specialist

IDEXX Laboratories, Inc.

tel 1-207-556-894 7

fax 1-207-556-4865

mobile 207-523-0835

patsy-root@idexx.com

idexx.com

# 06-XXXXX-00 

# Colilert ® 

# -18

**DRAFT**

# Colilert ® 

# -18 Test Kit

Introduction

Colilert-18 either simultaneously detects total coliforms and E. coli or fecal

coliforms in water. It is based on IDEXX’s patented Defined Substrate Technology ®



# (DST ® 

# ). When total/fecal coliforms metabolize Colilert-18’s nutrient-indicator,

ONPG, the sample turns yellow. When E. colimetabolize Colilert-18’s nutrientindicator, MUG, the sample fluoresces. Colilert-18 can detect these bacteria at 1

cfu/100 ml within 18 hours even with as many as 2 million heterotrophic bacteria

per 100 ml present.



# Contents

• WP020I-18 contains 20 Snap Packs for 100 mL samples

• WP200I-18 contains 200 Snap Packs for 100 mL samples



# Storage

Store at 2°C–25°C away from light.



# Presence/Absence (P/A) Procedure

1. Add contents of one pack to a 100 mL sample in a sterile, tran sparent,

nonfluorescing vessel.

2. Cap vessel and shake.

3. If sample is not already at 33°C–38°C, then place vessel in a 35°C waterbath for

20 minutes or, alternatively, a 44.5°C waterbath for 7–10 minutes.

4. Incubate at 35°C ±0.5°C for the remainder of the 18 hours.

5. Read results according to Result Interpretation table below.



# Quanti-Tray ® 

# Enumeration Procedure

1. Add contents of one

pack to a 100 mL room

temperature water sample

in a sterile vessel.

2. Cap vessel and shake

until dissolved.

3. Pour sample/reagent mixture into a

Quanti-Tray or Quanti-Tray/2000 and

seal in an IDEXX Quanti-Tray Sealer.

# 4. Place the sealed tray in a 35°C ±0.5°C (or 44.5°C ±0.2°C for fecal coliforms)

incubator for 18 hours (prewarming is not required).

5. Read results according to the Result Interpretation table below. Count the

number of positive wells and refer to the MPN table provided with the trays to

obtain a Most Probable Number.



# Result Interpretation

Appearance Result



# Less yellow than

the comparator * 

# Negative for total coliforms,

E. coli and fecal coliforms

Yellow equal to or greater than

the comparator * 

# when incubated

at 35°C ±0.5°C Positive for total coliforms

Yellow and fluorescence equal to

or greater than the comparator *



# when incubated at 35°C ±0.5°C Positive for E. coli

Yellow equal to or greater than

the comparator * 

# when incubated

at 44.5°C ±0.2°C Positive for fecal coliforms

• Look for fluorescence with a 6-watt, 365-nm UV light within 5 inches of the

sample in a dark environment. Face light away from your eyes and

towards the sample.

• Colilert-18 results are definitive at 18–22 hours. In addition, positives for both

total coliforms and E. coli observed before 18 hours and negatives observed

after 22 hours are also valid.



# Procedural Notes

• A slight tinge may be observed when Colilert-18 is added to the sample.

• If excess foam causes problems while using Quanti-Tray, you may choose to

use IDEXX Antifoam Solution (Catalog# WAFDB) or IDEXX 120 ml vessels with

Antifoam (Catalog# WV120SBAF-200).

• When following AFNOR regulations for drinking water testing , place the sealed

Quanti-Tray in a 36°C ±2°C incubator for 18 hours (prewarming to 36°C is not

required).

• This insert may not reflect your local regulations. For compliance testing, be

sure to follow appropriate regulatory procedures.

# • Colilert-18 can be run in any multiple tube format. Standard Methods for the

Examination of Water and Wastewater MPN tables should be used to find Most

Probable Numbers (MPNs).

• If a water sample has some background color, compare inoculated Colilert-18

sample to a control blank of the same water sample.

• Colilert-18 can be used for E. coli (but not coliforms) in marine water. Samples

must be diluted at least tenfold. Multiply the MPN value by the dilution factor to

obtain the proper quantitative result.

• Use only sterile, nonbuffered, oxidant-free water for dilutions.

• Colilert-18 is a primary water test. Colilert-18 performance characteristics do

not apply to samples altered by any pre-enrichment or concentration.

• In samples with excessive chlorine, a blue flash may be seen whe n adding

Colilert-18. If this is seen, consider sample invalid and discontinue testing.

• Aseptic technique should always be followed when using Colilert-18. Dispose

of in accordance with Good Laboratory Practices.



# Quality Control Procedures

The following quality control procedure is recommended for each lot of Colilert-18:

1. Fill three sterile vessels with 100 mL sterile water and inocul ate one with an

E. coli, one with a total coliform and one with a non-coliform using one of the

following products:

A. Quanti-Cult TM3 

# E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa

B. EZ-QC ®4 

# E. coli, Klebsiella oxytoca and Pseudomonas aeruginosa

C. A sterile loop of ATCC 5 

# strains, E. coli ATCC 25922 or 11775, Klebsiella

pneumoniae ATCC 31488 and Pseudomonas aeruginosa ATCC 10145 or

27853

2. Follow the P/A Procedure or Quanti-Tray Enumeration Procedure above.

3. Results should match the Result Interpretation table above.

**Note:** For fecal coliforms, EZ-QC E. Coli and Pseudomonas aeruginosa should be

used for quality control.



# For IDEXX Technical Support, please call:

North/South America: 207-556-4630/1-800-321-0207

Europe: 00800 4339 9111

UK: +44 (0) 1638 676800

China: +86-21-61279528

Japan: +81-422-71-5921

Australia: 1 800 655 978

# 1 

# IDEXX P/A Comparator, catalog # WP104; Quanti-Tray Comparator #WQTC, or Quanti-Tray/2000 Comparator

#WQT2KC

2 

# Eaton AD, Clesceri LS, Greenberg AE. Standard Methods for the Examination of Water and Wastewater . American

Public Health Association, 1998, Washington, DC.

3 

# Quanti-CultTM cultures - IDEXX catalog # WKIT-1001

4 

# EZ-QC Cultures – IDEXX Catalog #WEZQ

5 

# American Type Culture Collection 1-800-638-6597

Colilert, Defined Substrate Technology, DST and Quanti-Tray are either trademarks or registered trademaks of IDEXX

Laboratories, Inc. in the United States and/or other countries. Quanti-Cult is a trademark of Remel Inc.

© 2010 IDEXX Laboratories, Inc. All rights reserved.

Manufactured under one or more of the following U.S. patents: 4,925,789; 5,429,933; 5,518,892; 5,610,029; 5,620,

865; 5,620,895; 5,753,456 and 5,780,259. Other U.S. and/or foreign patents issued or pending.

# IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

www.idexx.com/water IDX 33/01 – 11/09 MÉTHODES ALTERNATIVES D’ANALYSE

Certifiée par AFNOR Certification

www.afnor.fr

La méthode IDEXX Colilert ® 

# -18/Quanti-Tray ® 

# a reçu le

droit d’usage de la Marque AFNOR Validation pour le

dénombrement des E. coli et des coliformes dans l’eau

de consommation humaine.

# 06-XXXXX-00 

# Colilert ®



# DRAFT

# Introduction

Colilert either simultaneously detects total coliforms and E. coli or fecal coliforms in

water. It is based on IDEXX’s patented Defined Substrate Technology ® 

# (DST ® 

# ). When

total coliforms metabolize Colilert’s nutrient-indicator, ONPG, the sample turns yellow.

When E. coli metabolize Colilert’s nutrient-indicator, MUG, the sample fluoresces.

Colilert can detect these bacteria at 1 cfu/100 mL within 24 hours even with as many

as 2 million heterotrophic bacteria per 100 mL present.



# Contents

WP020I contains 20 Snap Packs for 100 mL samples •

WP200I contains 200 Snap Packs for 100 mL samples •



# Storage

Store at 2°C–30°C away from light.



# Presence/Absence (P/A) Procedure

1. Add contents of one pack to a 100 mL sample in a sterile, transparent,

nonfluorescing vessel.

2. Cap vessel and shake.

3. Incubate at 35°C ± 0.5°C for 24 hours.

4. Read results according to Result Interpretation table below.



# Quanti-Tray ® 

# Enumeration Procedure

1. Add contents of one pack to

a 100 mL water sample in a

sterile vessel.

2. Cap vessel and shake until

dissolved.

3. Pour sample/reagent mixture into a

Quanti-Tray or Quanti-Tray/2000 and

seal in an IDEXX Quanti-Tray Sealer. 

# Colilert ® 

# Test Kit

# 4. Place the sealed tray in a 35°C ± 0.5°C (or 44.5°C ± 0.2°C for fecal coliforms)

incubator for 24 hours.

5. Read results according to the Result Interpretation table below. Count the number

of positive wells and refer to the MPN table provided with the trays to obtain a Most

Probable Number.



# Result Interpretation

Appearance Result



# Less yellow than

the comparator * 

# Negative for total coliforms,

E. coli and fecal coliforms

Yellow equal to or greater than

the comparator * 

# when incubated

at 35°C ±0.5°C Positive for total coliforms

Yellow and fluorescence equal to

or greater than the comparator *



# when incubated at 35°C ±0.5°C Positive for E. coli

Yellow equal to or greater than

the comparator * 

# when incubated

at 44.5°C ±0.2°C Positive for fecal coliforms

• Look for fluorescence with a 6-watt, 365 nm UV light within 5 inches of the sample

in a dark environment. Face light away from your eyes and towards the sample.

• Colilert results are definitive at 24–28 hours. In addition, positives for both total

coliforms and E. coli observed before 24 hours and negatives observed after 28

hours are also valid.



# Procedural Notes

•	 This insert may not reflect your local regulations. For compliance testing, be sure

to follow appropriate regulatory procedures.

• Colilert can be run in any multiple tube format. Standard Methods for the

Examination of Water and Wastewater 2 

# MPN tables should be used to find Most

Probable Numbers (MPNs).

• If a water sample has some background color, compare inoculated Colilert

sample to a control blank of the same water sample.

• If sample dilutions are made, multiply the MPN value by the dilution factor to

obtain the proper quantitative result.

# • Use only sterile, nonbuffered, oxidant-free water for dilutions.

• Colilert is a primary water test. Colilert performance characteristics do not apply to

samples altered by any pre-enrichment or concentration.

• In samples with excessive chlorine, a blue flash may be seen when adding

Colilert. If this is seen, consider sample invalid and discontinue testing.

• Aseptic technique should always be followed when using Colilert. Dispose of in

accordance with Good Laboratory Practices.



# Quality Control Procedures

The following quality control procedure is recommended for each lot of Colilert:

1. Fill three sterile vessels with 100 mL sterile water and inoculate one with an

E. coli, one with a total coliform and one with a non-coliform using one of the

following products:

A. Quanti-Cult TM3 

# E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa

B. EZ-QC ®4 

# E. coli, Klebsiella oxytoca and Pseudomonas aeruginosa

C. A sterile loop of ATCC 5 

# strains, E. coli ATCC 25922 or 11775, Klebsiella

pneumoniae ATCC 31488 and Pseudomonas aeruginosa ATCC 10145 or

27853

2. Follow the P/A Procedure or Quanti-Tray Enumeration Procedure above.

3. Results should match the Result Interpretation table above.

**Note:** For fecal coliforms, EZ-QC E. Coli and Pseudomonas aeruginosa should be used

for quality control.



# For IDEXX Technical Support, please call:

North/South America: 207-556-4630/1-800-321-0207

Europe: 00800 4339 9111

UK: +44 (0) 1638 676800

China: +86-21-61279528

Japan: +81-422-71-5921

Australia: 1 800 655 978

# 1 

# IDEXX P/A Comparator, catalog #WP104; Quanti-Tray Comparator #WQTC, or Quanti-Tray/2000 Comparator #WQT2KC

2 

# Eaton AD, Clesceri LS, Greenberg AE. Standard Methods for the Examination of Water and Wastewater . American Public

Health Association, 1998, Washington, DC.

3 

# Quanti-Cult cultures – IDEXX catalog # WKIT-1001

4 

# EZ-QC Cultures – IDEXX Catalog #WEZQ

5 

# American Type Culture Collection 1-800-638-6597

Colilert, Defined Substrate Technology, DST and Quanti-tray are either trademarks or registered trademarks of IDEXX

Laboratories, Inc. in the United States and/or other countries. Quanti-Cult is a trademark of Remel Inc.

© 2010 IDEXX Laboratories, Inc. All rights reserved.

Manufactured under one or more of the following U.S. patents: 4,925,789; 5,429,933; 5,518,892; 5,610,029; 5,620, 865;

5,620,895; 5,753,456 and 5,780,259. Other U.S. and/or foreign patents issued or pending.

# IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

www.idexx.com/water

# IDEXX Quanti-Tray

Insert and Most Probable Number (MPN) Table

06-02030-12 IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA



# www.idexx.com/water This certifies that the enclosed Quanti-Trays have

been sterilized with ethylene oxide.

For further information or documentation, contact

IDEXX Laboratories, Inc.

North/South America: 1-207-556-4496/1-800-321-0207

Europe: 00800 4339 9111

UK: +44 (0) 1638 676800

China: +86-21-61279528

Japan: +81 422 71 5921

Australia: 1800 655 978 

# Quanti-Tray Certificate of Sterility

# IDEXX Quanti-Tray

English Version

Quanti-Tray and Defined Substrate Technology are trademarks or registered trademarks of IDEXX Laboratories, Inc. in the

United States and/or other countries. Covered by U.S. Patent Nos. 5,518,892; 5,620,895; and/or 5,753,456. Other patents pending. 

# Introduction

IDEXX Quanti-Trays are designed to give quantitated bacterial counts of 100 mL samples using IDEXX Defined Substrate Technology

reagent products. Add the reagent/sample mixture to a Quanti-Tray, seal it in a Quanti-Tray Sealer and incubate per the reagent

instructions. Then count the number of positive wells and use the Most Probable Number (MPN) Table attached to determine the MPN.



# Contents

This package contains sterile 51-well Quanti-Trays.



# For Technical Assistance

**U.S./Canada 1-800-321-0207 or 1-207-556-4496**

**www.idexx.com/water** 

# User Instructions

1. Use one hand to hold a Quanti-Tray

upright with the well side facing

the palm. 2. Squeeze the upper part of the QuantiTray so that the Quanti-Tray bends

toward the palm. 3. Open the Quanti-Tray by pulling the

foil tab away from the well side. Avoid

touching the inside of the foil or tray.

4. Pour the reagent/sample mixture

directly into the Quanti-Tray, avoiding

contact with the foil tab. Allow foam

to settle. 5. Place the sample-filled Quanti-Tray

onto the rubber tray carrier of the

Quanti-Tray Sealer with the well side

(plastic) of the Quanti-Tray facing

down to fit into the carrier. 6. Seal according to the Quanti-Tray

Sealer instructions.

7. Incubate according to reagent

instructions.

8. Count positive wells and refer to

the MPN table on the back of this

instruction sheet to find the MPN.

9. Dispose of media in accordance with

good laboratory practices.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

# 51-Well Quanti-Tray MPN Table

No. of wells giving

positive reaction per

100 mL sample Most Probable

Number (MPN) 95% Confidence Limits

Lower Upper

0 <1 .0 0.0 3.7

1 1.0 0.3 5.6

2 2.0 0.6 7.3

3 3.1 1.1 9.0

4 4.2 1.7 10.7

5 5.3 2.3 12.3

6 6.4 3.0 13.9

7 7.5 3.7 15.5

8 8.7 4.5 17.1

9 9.9 5.3 18.8

10 11.1 6.1 20.5

11 12.4 7.0 22.1

12 13.7 7.9 23.9

13 15.0 8.8 25.7

14 16.4 9.8 27.5

15 17.8 10.8 29.4

16 19.2 11.9 31.3

17 20.7 13.0 33.3

18 22.2 14.1 35.2

19 23.8 15.3 37.3

20 25.4 16.5 39.4

21 27.1 17.7 41.6

22 28.8 19.0 43.9

23 30.6 20.4 46.3

24 32.4 21.8 48.7

25 34.4 23.3 51.2

26 36.4 24.7 53.9

27 38.4 26.4 56.6

28 40.6 28.0 59.5

29 42.9 29.7 62.5

30 45.3 31.5 65.6

31 47.8 33.4 69.0

32 50.4 35.4 72.5

33 53.1 37.5 76.2

34 56.0 39.7 80.1

35 59.1 42.0 84.4

36 62.4 44.6 88.8

37 65.9 47.2 93.7

38 69.7 50.0 99.0

39 73.8 53.1 104.8

40 78.2 56.4 111.2

41 83.1 59.9 118.3

42 88.5 63.9 126.2

43 94.5 68.2 135.4

44 101.3 73.1 146.0

45 109.1 78.6 158.7

46 118.4 85.0 174.5

47 129.8 92.7 195.0

48 144.5 102.3 224.1

49 165.2 115.2 272.2

50 200.5 135.8 387.6

51 > 200.5 146.1 infinite

# IDEXX Quanti-Tray

Version Française

Quanti-Tray et Defined Substrate Technology sont des marques de fabrique ou des marques déposées d'IDEXX Laboratories, Inc. aux États-Unis et/ou dans d'autres pays. Protégé par les brevets américains

n° 5,518,892; 5,620,895; and/or 5,753,456. Autres brevets en cours d'enregistrement. 

# Introduction

Les plateaux Quanti-Tray d’IDEXX, spécialement conçus pour des numérations bactériennes sur des échantillons de 100 millilitres, font

appel à la technologie Defined Substrate Technology d’IDEXX basée sur l’utilisation de réactifs. Versez le mélange réactif/échantillon

dans un Quanti-Tray, placez ensuite le tout dans le Quanti-Tray Sealer et procédez à l’incubation du mélange selon les instructions

accompagnant les réactifs. Comptez ensuite le nombre de cupules positives et référez-vous à la table ci-dessous pour déterminer le

nombre le plus probable (NPP).



# Contentu

Cette boîte contient 100 Quanti-Tray stériles composés chacun de 51 cupules.



# Services techniques IDEXX

**00800-4339-9111**

**www.idexx.fr/eau** 

# Mode d'emploi

1. Prenez un Quanti-Tray et tenez-le à

la verticale en veillant à orienter les

cupules vers la paume de la main. 2. Appuyez sur la partie supérieure du

Quanti-Tray afin qu’il vienne se loger

dans la paume de la main. 3. Ouvrez le Quanti-Tray en écartant la

languette en aluminium des cupules.

Evitez tout contact avec la partie

intérieure de l’aluminium ou du plateau.

4. Versez directement le mélange

réactif/échantillon dans le QuantiTray en évitant tout contact avec la

languette d’aluminium. Laisser la

mousse se dissiper. 5. Placez le Quanti-Tray rempli

d’échantillons sur le porte-plateau en

caoutchouc du Quanti-Tray Sealer en

veillant à orienter les cupules (parties

en plastique) vers le bas afin qu’elles

viennent s’encastrer parfaitement

dans le porte-plateau. 6. Scellez le Quanti-Tray conformément

aux instructions accompagnant le

Sealer.

7. Incubez conformément aux

instructions accompagnant le réactif.

8. Comptez le nombre de cupules

positives, puis référez-vous à la table

NPP au verso pour déterminer le

nombre le plus probable (NPP).

9. Débarrassez-vous du milieu en suivant

les bonnes pratiques de laboratoire.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

# Table NPP du Quanti-Tray a “51 Cupules”

Nombre de cupules

donnant une

réaction positive par

échantillon de 100 ml Nombre le plus

probable Limites de confiance à 95%

Minimum Maximum

0 <1 .0 0.0 3.7

1 1.0 0.3 5.6

2 2.0 0.6 7.3

3 3.1 1.1 9.0

4 4.2 1.7 10.7

5 5.3 2.3 12.3

6 6.4 3.0 13.9

7 7.5 3.7 15.5

8 8.7 4.5 17.1

9 9.9 5.3 18.8

10 11.1 6.1 20.5

11 12.4 7.0 22.1

12 13.7 7.9 23.9

13 15.0 8.8 25.7

14 16.4 9.8 27.5

15 17.8 10.8 29.4

16 19.2 11.9 31.3

17 20.7 13.0 33.3

18 22.2 14.1 35.2

19 23.8 15.3 37.3

20 25.4 16.5 39.4

21 27.1 17.7 41.6

22 28.8 19.0 43.9

23 30.6 20.4 46.3

24 32.4 21.8 48.7

25 34.4 23.3 51.2

26 36.4 24.7 53.9

27 38.4 26.4 56.6

28 40.6 28.0 59.5

29 42.9 29.7 62.5

30 45.3 31.5 65.6

31 47.8 33.4 69.0

32 50.4 35.4 72.5

33 53.1 37.5 76.2

34 56.0 39.7 80.1

35 59.1 42.0 84.4

36 62.4 44.6 88.8

37 65.9 47.2 93.7

38 69.7 50.0 99.0

39 73.8 53.1 104.8

40 78.2 56.4 111.2

41 83.1 59.9 118.3

42 88.5 63.9 126.2

43 94.5 68.2 135.4

44 101.3 73.1 146.0

45 109.1 78.6 158.7

46 118.4 85.0 174.5

47 129.8 92.7 195.0

48 144.5 102.3 224.1

49 165.2 115.2 272.2

50 200.5 135.8 387.6

51 > 200.5 146.1 infini

# IDEXX Quanti-Tray

Version Italiano

Quanti-Tray e Defined Substrate Technology sono marchi di proprietà di, e/o registrati da, IDEXX Laboratories, Inc.

e protetti negli Stati Uniti e/o in altri paesi. Protetto dai brevetti degli U.S.A. nn. 5,518,892; 5,620,895; and/or 5,753,456.

Altre domande di brevetto pendenti. 

# Introduzione

Quanti-Tray IDEXX sono concepiti per fornire la densità batterica (quantitativa) di campioni da 100 mL utilizzando reagenti Defined

Substrate Technology IDEXX. Aggiungere la miscela di reagente/campione ad un Quanti-Tray, inserire quest’ultimo nel Quanti-Tray Sealer

ed incubare in conformità alle istruzioni fornite con il reagente. Contare quindi il numero di pozzetti positivi e, con l’ausilio della tabella

MPN allegata, determinare il numero più probabile (MPN).



# Contenuto

Questa confezione contiene Quanti-Tray sterili, a 51 pozzetti.



# Supporto Tecnico IDEXX

**00800-4339-9111**

**www.idexx.it/acqua** 

# Istruzioni per l’uso

1. Con una mano, tenere il Quanti-Tray

in posizione verticale, con il lato dei

pozzetti rivolto verso il palmo. 2. Comprimere la parte superiore del

Quanti-Tray in modo da fletterlo verso

il palmo. 3. Aprire il Quanti-Tray tirando la linguetta

in alluminio in direzione opposta al lato

dei pozzetti. Evitare di toccare la parte

interna del foglio protettivo in alluminio

o del dispositivo a pozzetti.

4. Versare direttamente la miscela di

reagente/campione nel Quanti-Tray,

evitando di toccare la linguetta in

alluminio. Consentire alla schiuma

di sedimentare. 5. Una volta versata la miscela, collocare

il Quanti-Tray nel portapozzetti in

gomma del Quanti-Tray Sealer, con

il lto dei pozzetti (in plastica) rivolto

verso il basso, in modo da assicurarne

il corretto posizionamento. 6. Sigillare secondo le istruzioni del

Quanti-Tray Sealer.

7. Incubare secondo le istruzioni fornite

con il reagente.

8. Contare i pozzetti positivi e consultare

la tabella MPN sul retro del foglio delle

istruzioni, per individuare il numero più

probabile. (MPN) corretto.

9. Eliminare el terreno in accordo alle

Buone pratiche di laboratorio.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

# Tabella MPN Quanti-Tray a 51 pozzetti

N° di pozzetti positivi

in un campione da

100 ml Numero piú

probabile Limiti fiduciali del

95% Inferiore Limiti fiduciali del

95% Superiore

0 <1 .0 0.0 3.7

1 1.0 0.3 5.6

2 2.0 0.6 7.3

3 3.1 1.1 9.0

4 4.2 1.7 10.7

5 5.3 2.3 12.3

6 6.4 3.0 13.9

7 7.5 3.7 15.5

8 8.7 4.5 17.1

9 9.9 5.3 18.8

10 11.1 6.1 20.5

11 12.4 7.0 22.1

12 13.7 7.9 23.9

13 15.0 8.8 25.7

14 16.4 9.8 27.5

15 17.8 10.8 29.4

16 19.2 11.9 31.3

17 20.7 13.0 33.3

18 22.2 14.1 35.2

19 23.8 15.3 37.3

20 25.4 16.5 39.4

21 27.1 17.7 41.6

22 28.8 19.0 43.9

23 30.6 20.4 46.3

24 32.4 21.8 48.7

25 34.4 23.3 51.2

26 36.4 24.7 53.9

27 38.4 26.4 56.6

28 40.6 28.0 59.5

29 42.9 29.7 62.5

30 45.3 31.5 65.6

31 47.8 33.4 69.0

32 50.4 35.4 72.5

33 53.1 37.5 76.2

34 56.0 39.7 80.1

35 59.1 42.0 84.4

36 62.4 44.6 88.8

37 65.9 47.2 93.7

38 69.7 50.0 99.0

39 73.8 53.1 104.8

40 78.2 56.4 111.2

41 83.1 59.9 118.3

42 88.5 63.9 126.2

43 94.5 68.2 135.4

44 101.3 73.1 146.0

45 109.1 78.6 158.7

46 118.4 85.0 174.5

47 129.8 92.7 195.0

48 144.5 102.3 224.1

49 165.2 115.2 272.2

50 200.5 135.8 387.6

51 > 200.5 146.1 Infinito

# IDEXX Quanti-Tray

Versíon Español

Quanti-Tray y Defined Substrate Technology son marcas o marcas registradas de IDEXX Laboratories, Inc. en los

Estados Unidos y/o otros paises. Patente norteamericanas numero 5,518,892; 5,620,895; and/or 5,753,456. Otras patentes

pendientes de concesión. 

# Introducción

Los dispositivos Quanti-Tray de IDEXX están diseñados para producir recuentos bacterianos cuantificados de muestras de 100 mL, al

ser utilizadas con productos de reactivos de la Tecnología del Sustrato Definido (Defined Substrate Technology) de IDEXX. Agregue la

mezcla de reactivo y muestra a un dispositivo Quanti-Tray, séllelo en el Quanti-Tray Sealer (Sellador) e incúbela según las instrucciones del

reactivo. Luego cuente el número de celdas positivas y utilice la tabla de NMP adjunta para determinar el Número Más Probable (NMP).



# Contenido

Este paquete contiene 100 dispositivos Quanti-Tray estériles, de 51 celdas c/u.



# Soporte técnico de IDEXX

**00800-4339-9111**

**www.idexx.es/agua** 

# Instrucciones para el usuario

1. Sostenga en una mano el dispositivo

Quanti-Tray en posición vertical, con

el lado de las celdas orientado hacia

la palma. 2. Apriete la parte superior de el

dispositivo Quanti-Tray de modo

de doblarla hacia la palma. 3. Abra el dispositivo Quanti-Tray

desprendiendo la lengüeta metálica del

lado que contiene las celdas. Evite tocar

el interior del metal o del dispositivo.

4. Vierta la mezcla de reactivo y la

muestra directamente dentro de el

dispositivo Quanti-Tray, evitando tocar

la lengüeta metálica. Deja reposar

la espuma. 5. Coloque el dispositivo Quanti-Tray

0lleno de muestra sobre el

portadispositivo de goma del sellador

Quanti-Tray, orientando el lado de las

celdas (plásticas) del dispositivo hacia

abajo de manera que quepa en el

portadispositivo. 6. Selle el dispositivo según las

instrucciones del sellador.

7. Incube de acuerdo con las

instrucciones del reactivo.

8. Cuente las celdas positivas. Para

determinar el número más probable,

recurra a la tabla NMP al dorso de

esta hoja de instrucciones.

9. Verter el medio de cultivo conforme a

Buenas Prácticas del Laboratorio.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

# Tabla de NMP para dispositivo Quanti-Tray de 51 celdas

No de celdas que

producen una

reacción positiva, por

muestra de 100 ml Número más

probable Límites de confianza del 95%

Inferior Superior

0 <1 .0 0.0 3.7

1 1.0 0.3 5.6

2 2.0 0.6 7.3

3 3.1 1.1 9.0

4 4.2 1.7 10.7

5 5.3 2.3 12.3

6 6.4 3.0 13.9

7 7.5 3.7 15.5

8 8.7 4.5 17.1

9 9.9 5.3 18.8

10 11.1 6.1 20.5

11 12.4 7.0 22.1

12 13.7 7.9 23.9

13 15.0 8.8 25.7

14 16.4 9.8 27.5

15 17.8 10.8 29.4

16 19.2 11.9 31.3

17 20.7 13.0 33.3

18 22.2 14.1 35.2

19 23.8 15.3 37.3

20 25.4 16.5 39.4

21 27.1 17.7 41.6

22 28.8 19.0 43.9

23 30.6 20.4 46.3

24 32.4 21.8 48.7

25 34.4 23.3 51.2

26 36.4 24.7 53.9

27 38.4 26.4 56.6

28 40.6 28.0 59.5

29 42.9 29.7 62.5

30 45.3 31.5 65.6

31 47.8 33.4 69.0

32 50.4 35.4 72.5

33 53.1 37.5 76.2

34 56.0 39.7 80.1

35 59.1 42.0 84.4

36 62.4 44.6 88.8

37 65.9 47.2 93.7

38 69.7 50.0 99.0

39 73.8 53.1 104.8

40 78.2 56.4 111.2

41 83.1 59.9 118.3

42 88.5 63.9 126.2

43 94.5 68.2 135.4

44 101.3 73.1 146.0

45 109.1 78.6 158.7

46 118.4 85.0 174.5

47 129.8 92.7 195.0

48 144.5 102.3 224.1

49 165.2 115.2 272.2

50 200.5 135.8 387.6

51 > 200.5 146.1 infinito

# IDEXX Quanti-Tray

Deutsch Version

Quanti-Tray und Defined Substrate Technology sind Schutzmarken oder eingetragene Schutzmarken von

IDEXX Laboratories, Inc. in den Vereinigten Staaten und/oder in anderen Ländern. Geschützt durch U.S.-Patent

Nummern 5,518,892; 5,620,895; and/or 5,753,456. Andere Patenten angemeldet. 

# Einführung

Quanti-Tray von IDEXX dienen zur quantitativen Bakterienzählung in 100-mL-Proben unter Verwendung von IDEXX Defined

Substrate Technology-Reagenzien. Die Mischung aus Probe und Reagenz in einen Quanti-Tray geben, diesen in einem Quanti-TrayVersiegelungsgerät verschließen und laut Reagenz-Packungsbeilage inkubieren. Anschließend die Anzahl der positiven Vertiefungen

zählen und mit der beigefügten MPN-Tabelle die “Wahrscheinlichste Anzahl” (MPN) bestimmen.



# Inhalt

Diese Packung enthält 100 sterile Quanti-Tray mit jeweils 51 Vertiefungen.



# IDEXX Technischer Dienst

**00800-4339-9111**

**www.idexx.de/wasser** 

# Gebrauchsanweisung

1. Quanti-Tray mit einer Hand fassen und

senkrecht halten, wobei die Seite mit

den Vertiefungen zur Handfläche zeigt. 2. Den oberen Teil des Quanti-Trays

zusammendrücken, so daß sich der

Träger zur Handfläche hin biegt. 3. Quanti-Tray öffnen, indem die

Folienlasche von der Seite mit den

Vertiefungen abgezogen wird. Die

Innenseite der Folie oder des Trays

nicht berühren.

4. Die Mischung aus Probe und

Reagenz direkt in einen Quanti-Tray

gießen. Dabei eine Berührung der

Folienlasche vermeiden. Schaum

einwirken lassen. 5. Den mit der Probe gefüllten QuantiTray auf die Gummi-Trägerunterlage

des Quanti-Tray-Versiegelungsgeräts

stellen. Die Seite mit den Vertiefungen

(Kunststoff) des Quanti-Trays muß

dabei nach unten weisen, damit sie in

die Unterlage paßt. 6. Das Verschließen erfolgt

laut Anweisungen für das

Versiegelungsgerät.

7. Entsprechend den

Reagenzanweisungen inkubieren.

8. Die positiven Vertiefungen zählen

und mit Hilfe der MPN-Tabelle auf der

Rückseite dieser Packungsbeilage

die Wahrscheinlichste Anzahl (MPN)

ermitteln.

9. Entsorgen Sie die Quanti-Trays gemäß

guter Laborpraxis.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

# MPN-Tabelle für Quanti-Tray mit 51 Vertiefungen

Anzahl der

Vertiefungen, die bei

einer 100-ml-Probe

positiv reagieren Wahrscheinlichste

Anzahl 95% Vertrauensgrenze

Untere Obere

0 <1 .0 0.0 3.7

1 1.0 0.3 5.6

2 2.0 0.6 7.3

3 3.1 1.1 9.0

4 4.2 1.7 10.7

5 5.3 2.3 12.3

6 6.4 3.0 13.9

7 7.5 3.7 15.5

8 8.7 4.5 17.1

9 9.9 5.3 18.8

10 11.1 6.1 20.5

11 12.4 7.0 22.1

12 13.7 7.9 23.9

13 15.0 8.8 25.7

14 16.4 9.8 27.5

15 17.8 10.8 29.4

16 19.2 11.9 31.3

17 20.7 13.0 33.3

18 22.2 14.1 35.2

19 23.8 15.3 37.3

20 25.4 16.5 39.4

21 27.1 17.7 41.6

22 28.8 19.0 43.9

23 30.6 20.4 46.3

24 32.4 21.8 48.7

25 34.4 23.3 51.2

26 36.4 24.7 53.9

27 38.4 26.4 56.6

28 40.6 28.0 59.5

29 42.9 29.7 62.5

30 45.3 31.5 65.6

31 47.8 33.4 69.0

32 50.4 35.4 72.5

33 53.1 37.5 76.2

34 56.0 39.7 80.1

35 59.1 42.0 84.4

36 62.4 44.6 88.8

37 65.9 47.2 93.7

38 69.7 50.0 99.0

39 73.8 53.1 104.8

40 78.2 56.4 111.2

41 83.1 59.9 118.3

42 88.5 63.9 126.2

43 94.5 68.2 135.4

44 101.3 73.1 146.0

45 109.1 78.6 158.7

46 118.4 85.0 174.5

47 129.8 92.7 195.0

48 144.5 102.3 224.1

49 165.2 115.2 272.2

50 200.5 135.8 387.6

51 > 200.5 146.1 endlos

# IDEXX Quanti-Tray

日本語版

Quanti-Tray および Defined Substrate Technology は米国および / または他国の IDEXX Laboratories, Inc. の商標または登録商標です。 以下の特許番

号の米国特許により保護されています。 5,518,892; 5,620,895; and/or 5,753,456. 他の特許は出願中です。 

# はじめに

IDEXX Quanti-Tray(QT- トレイ)は、 IDEXX が特許を取得している Defined Substrate Technology ( 特定酵素基質法)に基づいた Colilert を用い、検体

100 mL 中の最確数を測定する新しいシステムです。 Colilert と検体の混合液を Quanti-Tray に加え、シーラーで封入し、取扱説明書に従い培養して

ください。その後、陽性ウェル数を数え、添付の最確数(MPN)表を用いて、MPNを測定します。



# 内容

このパッケージには、滅菌済みの 51 ウェル Quanti-Tray が 100個入っています。



# ご質問やサポートに関しては以下にご連絡ください。

アイデックス ラボラトリーズ株式会社

TEL: 0422-71-5921 www.idexx.co.jp/water 

# 操作手順

1. ウェル側を手のひらに向けてトレイを垂直

に持ってください。 2. トレイが手のひらに向かって曲がるよう

に、トレイの上部を強く握ってください。 3. トレイのつまみをウェル側から引き離し

て、注入口を広げてください。トレイのつ

まみ、またはトレイの内部に触れないよう

にしてください。

4. トレイのつまみにかからないように直接、

Colilert と検体の混合液を注いでくださ

い。気泡は軽く指ではじくように取り除い

てください。 5. 検体の入ったトレイをシーラーのゴム製シ

ートの上にウェル側を下に向けてセットし

てください。 6. シーラーの取扱説明書に従って密封して

ください。

7. Colilert の取扱説明書に従って培養してく

ださい。

8. 陽性ウェル数を数え、添付の最確数

(MPN) 表を使用してMPNを求めてくだ

さい。

9. 検体及びトレイの廃棄は、高圧滅菌してく

ださい。

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA

# 51 ウェル **Quanti-Tray MPN** 表

100ml の検体で陽性

反応を呈するウェル数 最確数 (MPN) 95% 信頼限界

下限 上限

0 <1 .0 0.0 3.7

1 1.0 0.3 5.6

2 2.0 0.6 7.3

3 3.1 1.1 9.0

4 4.2 1.7 10.7

5 5.3 2.3 12.3

6 6.4 3.0 13.9

7 7.5 3.7 15.5

8 8.7 4.5 17.1

9 9.9 5.3 18.8

10 11.1 6.1 20.5

11 12.4 7.0 22.1

12 13.7 7.9 23.9

13 15.0 8.8 25.7

14 16.4 9.8 27.5

15 17.8 10.8 29.4

16 19.2 11.9 31.3

17 20.7 13.0 33.3

18 22.2 14.1 35.2

19 23.8 15.3 37.3

20 25.4 16.5 39.4

21 27.1 17.7 41.6

22 28.8 19.0 43.9

23 30.6 20.4 46.3

24 32.4 21.8 48.7

25 34.4 23.3 51.2

26 36.4 24.7 53.9

27 38.4 26.4 56.6

28 40.6 28.0 59.5

29 42.9 29.7 62.5

30 45.3 31.5 65.6

31 47.8 33.4 69.0

32 50.4 35.4 72.5

33 53.1 37.5 76.2

34 56.0 39.7 80.1

35 59.1 42.0 84.4

36 62.4 44.6 88.8

37 65.9 47.2 93.7

38 69.7 50.0 99.0

39 73.8 53.1 104.8

40 78.2 56.4 111.2

41 83.1 59.9 118.3

42 88.5 63.9 126.2

43 94.5 68.2 135.4

44 101.3 73.1 146.0

45 109.1 78.6 158.7

46 118.4 85.0 174.5

47 129.8 92.7 195.0

48 144.5 102.3 224.1

49 165.2 115.2 272.2

50 200.5 135.8 387.6

51 > 200.5 146.1 以上

# IDEXX Quanti-Tray/2000

Insert and Most Probable Number (MPN) Table

06-02320-10 IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA This certifies that the enclosed Quanti-Trays have

been sterilized with ethylene oxide.

For further information or documentation, contact

IDEXX Laboratories, Inc.

North/South America: 1-207-556-4496/1-800-321-0207

Europe: 00800 4339 9111

UK: +44 (0) 1638 676800

China: +86-21-61279528

Japan: +81 422 71 5921

Australia: 1800 655 978 

# Quanti-Tray Certificate of Sterility

# IDEXX Quanti-Tray/2000

English Version

Quanti-Tray and Defined Substrate Technology are trademarks or registered trademarks of IDEXX Laboratories, Inc. in the

United States and/or other countries. Covered by U.S. Patent Numbers 5,518,892; 5,620,895; and 5,753,456. Other patents pending. 

# Introduction

IDEXX Quanti-Tray/2000 is designed to give quantitated bacterial counts of 100 mL samples using IDEXX Defined Substrate Technology

reagent products. Add the reagent/sample mixture to a Quanti-Tray/2000, seal it in a Quanti-Tray Sealer and incubate per the reagent

instructions. Then count the number of positive large and small wells and use the Most Probable Number (MPN) Table attached to

determine the MPN.



# Contents

This package contains sterile Quanti-Tray/2000s.



# For Technical Support

**U.S./Canada 1-800-321-0207 or 1-207-556-4496**

**www.idexx.com/water** 

# User Instructions

1. Use one hand to hold a Quanti-Tray

upright with the well side facing

the palm. 2. Squeeze the upper part of the QuantiTray so that the Quanti-Tray bends

toward the palm. 3. Gently pull foil tab to separate the

foil from the tray. Avoid touching the

inside of the foil or tray.

4. Pour the reagent/sample mixture

directly into the Quanti-Tray, avoiding

contact with the foil tab. Tap the small

wells 2–3 times to release any air

bubbles. Allow foam to settle. 5. Place the sample-filled Quanti-Tray

onto the Quanti-Tray/2000 rubber

insert of the Quanti-Tray Sealer with

the well side (plastic) of the QuantiTray facing down. 6. Seal according to the Quanti-Tray

Sealer instructions.

7. Incubate according to reagent

instructions.

8. Count large and small positive wells

and refer to the Quanti-Tray/2000 MPN

table to find the MPN.

9. Dispose of media in accordance with

good laboratory practices.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA**IDEXX Quanti-Tray/2000 MPN Table # Large** **Wells Positive** **# Small Wells Positive 0** **1 2** **3 4** **5 6** **7 8** **9 10** **11 12** **13 14** **15 16** **17 18** **19 20** **21 22** **23 24** 0<11.02.03.04.05.06.07.08.09.010.011.012.013.014.115.116.117.118.119.120.221.222.223.324.311.02.03.04.05.06.07.18.19.110.111.112.113.214.215.216.217.318.319.320.421.422.423.524.525.622.03.04.15.16.17.18.19.210.211.212.213.314.315.416.417.418.519.520.621.622.723.724.825.826.933.14.15.16.17.28.29.210.311.312.413.414.515.516.517.618.619.720.821.822.923.925.026.127.128.244.15.26.27.28.39.310.411.412.513.514.615.616.717.818.819.921.022.023.124.225.326.327.428.529.655.26.37.38.49.410.511.512.613.714.715.816.917.919.020.121.222.223.324.425.526.627.728.829.931.066.37.48.49.510.611.612.713.814.916.017.018.119.220.321.422.523.624.725.826.928.029.130.231.332.477.58.59.610.711.812.813.915.016.117.218.319.420.521.622.723.824.926.027.128.329.430.531.632.833.988.69.710.811.913.014.115.216.317.418.519.620.721.822.924.125.226.327.428.629.730.832.033.134.335.499.810.912.013.114.215.316.417.618.719.820.922.023.224.325.426.627.728.930.031.232.333.534.635.837.01011.012.113.214.415.516.617.718.920.021.122.323.424.625.726.928.029.230.331.532.733.835.036.237.438.61112.213.414.515.616.817.919.120.221.422.523.724.826.027.228.329.530.731.933.034.235.436.637.839.040.21213.514.615.816.918.119.320.421.622.823.925.126.327.528.629.831.032.233.434.635.837.038.239.540.741.91314.816.017.118.319.520.621.823.024.225.426.627.829.030.231.432.633.835.036.237.538.739.941.242.443.61416.117.318.519.720.922.123.324.525.726.928.129.330.531.733.034.235.436.737.939.140.441.642.944.245.41517.518.719.921.122.323.524.725.927.228.429.630.932.133.334.635.837.138.439.640.942.243.444.746.047.31618.920.121.322.623.825.026.227.528.730.031.232.533.735.036.337.538.840.141.442.744.045.346.647.949.21720.321.622.824.125.326.627.829.130.331.632.934.135.436.738.039.340.641.943.244.545.947.248.549.851.21821.823.124.325.626.928.129.430.732.033.334.635.937.238.539.841.142.443.845.146.547.849.250.551.953.21923.324.625.927.228.529.831.132.433.735.036.337.639.040.341.643.044.345.747.148.449.851.252.654.055.42024.926.227.528.830.131.532.834.135.436.838.139.540.842.243.644.946.347.749.150.551.953.354.756.157.62126.527.929.230.531.833.234.535.937.338.640.041.442.844.145.546.948.449.851.252.654.155.556.958.459.92228.229.530.932.333.635.036.437.739.140.541.943.344.846.247.649.050.551.953.454.856.357.859.360.862.32329.931.332.734.135.536.838.339.741.142.543.945.446.848.349.751.252.754.255.657.158.660.261.763.264.72431.733.134.535.937.338.840.241.743.144.646.047.549.050.552.053.555.056.558.059.561.162.664.265.867.32533.635.036.437.939.340.842.243.745.246.748.249.751.252.754.355.857.358.960.562.063.665.266.868.470.02635.536.938.439.941.442.844.345.947.448.950.452.053.555.156.758.259.861.463.064.766.367.969.671.272.92737.438.940.442.043.545.046.548.149.651.252.854.456.057.659.260.862.464.165.767.469.170.872.574.275.92839.541.042.644.145.747.348.850.452.053.655.256.958.560.261.863.565.266.968.670.372.073.775.577.379.02941.743.244.846.448.049.651.252.854.556.157.859.561.262.964.666.368.069.871.573.375.176.978.780.582.43043.945.547.148.750.452.053.755.457.158.860.562.264.065.767.569.371.072.974.776.578.380.282.184.085.93146.247.949.551.252.954.656.358.159.861.663.365.166.968.770.572.474.276.178.079.981.883.785.787.689.63248.750.452.153.855.657.359.160.962.764.566.368.270.071.973.875.777.679.581.583.585.487.589.591.593.63351.253.054.856.558.360.262.063.865.767.669.571.473.375.277.279.281.283.285.287.389.391.493.695.797.83453.955.757.659.461.363.165.067.068.970.872.874.876.878.880.882.985.087.189.291.493.595.797.9100.2102.43556.858.660.562.464.466.368.370.372.374.376.378.480.582.684.786.989.191.393.595.798.0100.3102.6105.0107.33659.861.763.765.767.769.771.773.875.978.080.182.384.586.788.991.293.595.898.1100.5102.9105.3107.7110.2112.73762.965.067.069.171.273.375.477.679.882.084.286.588.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.9118.63866.368.470.672.774.977.179.481.683.986.288.691.093.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.2125.03970.072.274.476.778.981.383.686.088.490.993.495.998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.2132.24073.876.278.580.983.385.788.290.893.395.998.5101.2103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.0140.34178.080.583.085.588.090.693.395.998.7101.4104.3107.1110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.9149.54282.685.287.890.593.296.098.8101.7104.6107.6110.6113.7116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.1160.24387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.1124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.2172.84493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.6133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.9188.24599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.6143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.2207.546106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.5156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.4233.347114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.4172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.5270.048123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.0193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.0328.249135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6435.2Quanti-Tray and Defined Substrate Technology are trademarks or registered trademarks of IDEXX Laboratories, Inc. in the United States and/or other countries. Covered by U.S. Patent Numbers 4,925,789; 5,429,933; 5,518,892. Other patents pending.**IDEXX Quanti-Tray/2000 MPN Table # Large** **Wells Positive** **# Small Wells Positive 25** **26 27** **28 29** **30 31** **32 33** **34 35** **36 37** **38 39** **40 41** **42 43** **44 45** **46 47** **48**025.326.427.428.429.530.531.532.633.634.735.736.837.838.940.041.042.143.144.245.346.347.448.549.5126.627.728.729.830.831.932.934.035.036.137.238.239.340.441.442.543.644.745.746.847.949.050.151.2227.929.030.031.132.233.234.335.436.537.538.639.740.841.943.044.045.146.247.348.449.550.651.752.8329.330.431.432.533.634.735.836.837.939.040.141.242.343.444.545.646.747.848.950.051.252.353.454.5430.731.832.833.935.036.137.238.339.440.541.642.843.945.046.147.248.349.550.651.752.954.055.156.3532.133.234.335.436.537.638.739.941.042.143.244.445.546.647.748.950.051.252.353.554.655.856.958.1633.534.735.836.938.039.240.341.442.643.744.846.047.148.349.450.651.752.954.155.256.457.658.759.9735.036.237.338.439.640.741.943.044.245.346.547.748.850.051.252.353.554.755.957.158.359.460.661.8836.637.738.940.041.242.343.544.745.947.048.249.450.651.853.054.155.356.557.759.060.261.462.663.8938.139.340.541.642.844.045.246.447.648.850.051.252.453.654.856.057.258.459.760.962.163.464.665.81039.740.942.143.344.545.746.948.149.350.651.853.054.255.556.757.959.260.461.762.964.265.466.767.91141.442.643.845.046.347.548.749.951.252.453.754.956.157.458.659.961.262.463.765.066.367.568.870.11243.144.345.646.848.149.350.651.853.154.355.656.858.159.460.762.063.264.565.867.168.469.771.072.41344.946.147.448.649.951.252.553.755.056.357.658.960.261.562.864.165.466.768.069.370.772.073.374.71446.748.049.350.551.853.154.455.757.058.359.660.962.363.664.966.367.668.970.371.673.074.475.777.11548.649.951.252.553.855.156.457.859.160.461.863.164.565.867.268.569.971.372.674.075.476.878.279.61650.551.853.254.555.857.258.559.961.262.664.065.366.768.169.570.972.373.775.176.577.979.380.882.21752.553.955.256.658.059.360.762.163.564.966.367.769.170.571.973.374.876.277.679.180.582.083.584.91854.656.057.458.860.261.663.064.465.867.268.670.171.573.074.475.977.378.880.381.883.384.886.387.81956.858.259.661.062.463.965.366.868.269.771.172.674.175.577.078.580.081.583.184.686.187.689.290.72059.060.461.963.364.866.367.769.270.772.273.775.276.778.279.881.382.884.485.987.589.190.792.293.82161.362.864.365.867.368.870.371.873.374.976.477.979.581.182.684.285.887.489.090.692.293.895.497.12263.865.366.868.369.871.472.974.576.177.679.280.882.484.085.687.288.990.592.193.895.597.198.8100.52366.367.869.471.072.574.175.777.378.980.582.283.885.487.188.790.492.193.895.597.298.9100.6102.4104.12468.970.572.173.775.377.078.680.381.983.685.286.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92571.773.375.076.678.380.081.783.385.186.888.590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.0111.92674.676.378.079.781.483.184.886.688.490.191.993.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22777.679.481.182.984.686.488.290.091.993.795.597.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.72880.882.684.486.388.189.991.893.795.697.599.4101.3103.3105.2107.2109.2111.2113.2115.2117.3119.3121.4123.5125.62984.286.187.989.891.793.795.697.599.5101.5103.5105.5107.5109.5111.6113.7115.7117.8120.0122.1124.2126.4128.6130.83087.889.791.793.695.697.699.6101.6103.7105.7107.8109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.43191.693.695.697.799.7101.8103.9106.0108.2110.3112.5114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.53295.797.899.9102.0104.2106.3108.5110.7113.0115.2117.5119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.9125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.8131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.4145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.3153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.2162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.4173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.1185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.8199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.4216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.8238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.1265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.3302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.3355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.8436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.3574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4866.4920.8980.41046.21119.91203.31299.71413.61553.11732.91986.32419.6>2419.6

# IDEXX Quanti-Tray/2000

Version Française

Quanti-Tray et Defined Substrate Technology sont des marques de fabrique ou des marques déposées d'IDEXX Laboratories, Inc. aux États-Unis et/ou dans d'autres pays. Protégé par les brevets américains

n° 5,518,892; 5,620,895; and 5,753,456.. Autres brevets en cours d'enregistrement. 

# Introduction

Les plateaux Quanti-Tray/2000 d’IDEXX, spécialement conçus pour effectuer des numérations bactériennes sur des échantillons de 100

millilitres, font appel à la technologie Defined Substrate Technology d’IDEXX basée sur l’utilisation de réactifs. Versez le mélange réactif/

échantillon dans un Quanti-Tray/2000, placez ensuite le tout dans le Quanti-Tray Sealer et procédez à l’incubation du mélange selon

les instructions accompagnant les réactifs. Comptez ensuite le nombre de cupules positives et référez-vous à la table ci-dessous pour

déterminer le nombre le plus probable (NPP).



# Contentu

Cette boîte contient Quanti-Tray/2000 stériles.



# Services techniques IDEXX

**00800-4339-9111**

**www.idexx.fr/eau** 

# Mode d'emploi

1. Prenez un Quanti-Tray et tenez-le à

la verticale en veillant à orienter les

cupules vers la paume de la main. 2. Appuyez sur la partie supérieure du

Quanti-Tray afin qu’il vienne se loger

dans la paume de la main. 3. Ouvrez le Quanti-Tray en retirant

la feuille en aluminium. Evitez tout

contact avec la partie intérieure de

l’aluminium ou du plateau.

4. Versez directement le mélange réactif/

échantillon dans le Quanti-Tray en

évitant tout contact avec la languette

d’aluminium. Tapoter les petits puits 2

à 3 fois afin de libérer les bulles d'air.

Laisser la mousse se dissiper. 5. Placez le Quanti-Tray rempli

d’échantillons sur le porte-plateau en

caoutchouc du Quanti-Tray Sealer en

veillant à orienter les cupules (parties

en plastique) vers le bas afin qu’elles

viennent s’encastrer parfaitement

dans le porte-plateau. 6. Scellez le Quanti-Tray conformément

aux instructions accompagnant

le Sealer.

7. Incubez conformément aux

instructions accompagnant le réactif.

8. Comptez le nombre de cupules

positives, puis référez-vous à la table

NPP au verso pour déterminer le

nombre le plus probable (NPP).

9. Débarrassez-vous du milieu en suivant

les bonnes pratiques de laboratoire.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA**Table NPP IDEXX Quanti-Tray/2000 # Grands** **Puits Positifs** **# Petits Puits Positifs 0** **1 2** **3 4** **5 6** **7 8** **9 10** **11 12** **13 14** **15 16** **17 18** **19 20** **21 22** **23 24** 0<11.02.03.04.05.06.07.08.09.010.011.012.013.014.115.116.117.118.119.120.221.222.223.324.311.02.03.04.05.06.07.18.19.110.111.112.113.214.215.216.217.318.319.320.421.422.423.524.525.622.03.04.15.16.17.18.19.210.211.212.213.314.315.416.417.418.519.520.621.622.723.724.825.826.933.14.15.16.17.28.29.210.311.312.413.414.515.516.517.618.619.720.821.822.923.925.026.127.128.244.15.26.27.28.39.310.411.412.513.514.615.616.717.818.819.921.022.023.124.225.326.327.428.529.655.26.37.38.49.410.511.512.613.714.715.816.917.919.020.121.222.223.324.425.526.627.728.829.931.066.37.48.49.510.611.612.713.814.916.017.018.119.220.321.422.523.624.725.826.928.029.130.231.332.477.58.59.610.711.812.813.915.016.117.218.319.420.521.622.723.824.926.027.128.329.430.531.632.833.988.69.710.811.913.014.115.216.317.418.519.620.721.822.924.125.226.327.428.629.730.832.033.134.335.499.810.912.013.114.215.316.417.618.719.820.922.023.224.325.426.627.728.930.031.232.333.534.635.837.01011.012.113.214.415.516.617.718.920.021.122.323.424.625.726.928.029.230.331.532.733.835.036.237.438.61112.213.414.515.616.817.919.120.221.422.523.724.826.027.228.329.530.731.933.034.235.436.637.839.040.21213.514.615.816.918.119.320.421.622.823.925.126.327.528.629.831.032.233.434.635.837.038.239.540.741.91314.816.017.118.319.520.621.823.024.225.426.627.829.030.231.432.633.835.036.237.538.739.941.242.443.61416.117.318.519.720.922.123.324.525.726.928.129.330.531.733.034.235.436.737.939.140.441.642.944.245.41517.518.719.921.122.323.524.725.927.228.429.630.932.133.334.635.837.138.439.640.942.243.444.746.047.31618.920.121.322.623.825.026.227.528.730.031.232.533.735.036.337.538.840.141.442.744.045.346.647.949.21720.321.622.824.125.326.627.829.130.331.632.934.135.436.738.039.340.641.943.244.545.947.248.549.851.21821.823.124.325.626.928.129.430.732.033.334.635.937.238.539.841.142.443.845.146.547.849.250.551.953.21923.324.625.927.228.529.831.132.433.735.036.337.639.040.341.643.044.345.747.148.449.851.252.654.055.42024.926.227.528.830.131.532.834.135.436.838.139.540.842.243.644.946.347.749.150.551.953.354.756.157.62126.527.929.230.531.833.234.535.937.338.640.041.442.844.145.546.948.449.851.252.654.155.556.958.459.92228.229.530.932.333.635.036.437.739.140.541.943.344.846.247.649.050.551.953.454.856.357.859.360.862.32329.931.332.734.135.536.838.339.741.142.543.945.446.848.349.751.252.754.255.657.158.660.261.763.264.72431.733.134.535.937.338.840.241.743.144.646.047.549.050.552.053.555.056.558.059.561.162.664.265.867.32533.635.036.437.939.340.842.243.745.246.748.249.751.252.754.355.857.358.960.562.063.665.266.868.470.02635.536.938.439.941.442.844.345.947.448.950.452.053.555.156.758.259.861.463.064.766.367.969.671.272.92737.438.940.442.043.545.046.548.149.651.252.854.456.057.659.260.862.464.165.767.469.170.872.574.275.92839.541.042.644.145.747.348.850.452.053.655.256.958.560.261.863.565.266.968.670.372.073.775.577.379.02941.743.244.846.448.049.651.252.854.556.157.859.561.262.964.666.368.069.871.573.375.176.978.780.582.43043.945.547.148.750.452.053.755.457.158.860.562.264.065.767.569.371.072.974.776.578.380.282.184.085.93146.247.949.551.252.954.656.358.159.861.663.365.166.968.770.572.474.276.178.079.981.883.785.787.689.63248.750.452.153.855.657.359.160.962.764.566.368.270.071.973.875.777.679.581.583.585.487.589.591.593.63351.253.054.856.558.360.262.063.865.767.669.571.473.375.277.279.281.283.285.287.389.391.493.695.797.83453.955.757.659.461.363.165.067.068.970.872.874.876.878.880.882.985.087.189.291.493.595.797.9100.2102.43556.858.660.562.464.466.368.370.372.374.376.378.480.582.684.786.989.191.393.595.798.0100.3102.6105.0107.33659.861.763.765.767.769.771.773.875.978.080.182.384.586.788.991.293.595.898.1100.5102.9105.3107.7110.2112.73762.965.067.069.171.273.375.477.679.882.084.286.588.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.9118.63866.368.470.672.774.977.179.481.683.986.288.691.093.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.2125.03970.072.274.476.778.981.383.686.088.490.993.495.998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.2132.24073.876.278.580.983.385.788.290.893.395.998.5101.2103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.0140.34178.080.583.085.588.090.693.395.998.7101.4104.3107.1110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.9149.54282.685.287.890.593.296.098.8101.7104.6107.6110.6113.7116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.1160.24387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.1124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.2172.84493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.6133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.9188.24599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.6143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.2207.546106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.5156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.4233.347114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.4172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.5270.048123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.0193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.0328.249135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6435.2Quanti-Tray et Defined Substrate Technology sont des marques de fabrique ou des marques déposées d'IDEXX Laboratories, Inc. aux États-Unis et/ou dans d'autres pays. Protégé par les brevets américains n° 4,925,789; 5,429,933; 5,518,892. Autres brevets en cours d'enregistrement.**Table NPP IDEXX Quanti-Tray/2000 # Grands** **Puits Positifs** **# Petits Puits Positifs 25** **26 27** **28 29** **30 31** **32 33** **34 35** **36 37** **38 39** **40 41** **42 43** **44 45** **46 47** **48**025.326.427.428.429.530.531.532.633.634.735.736.837.838.940.041.042.143.144.245.346.347.448.549.5126.627.728.729.830.831.932.934.035.036.137.238.239.340.441.442.543.644.745.746.847.949.050.151.2227.929.030.031.132.233.234.335.436.537.538.639.740.841.943.044.045.146.247.348.449.550.651.752.8329.330.431.432.533.634.735.836.837.939.040.141.242.343.444.545.646.747.848.950.051.252.353.454.5430.731.832.833.935.036.137.238.339.440.541.642.843.945.046.147.248.349.550.651.752.954.055.156.3532.133.234.335.436.537.638.739.941.042.143.244.445.546.647.748.950.051.252.353.554.655.856.958.1633.534.735.836.938.039.240.341.442.643.744.846.047.148.349.450.651.752.954.155.256.457.658.759.9735.036.237.338.439.640.741.943.044.245.346.547.748.850.051.252.353.554.755.957.158.359.460.661.8836.637.738.940.041.242.343.544.745.947.048.249.450.651.853.054.155.356.557.759.060.261.462.663.8938.139.340.541.642.844.045.246.447.648.850.051.252.453.654.856.057.258.459.760.962.163.464.665.81039.740.942.143.344.545.746.948.149.350.651.853.054.255.556.757.959.260.461.762.964.265.466.767.91141.442.643.845.046.347.548.749.951.252.453.754.956.157.458.659.961.262.463.765.066.367.568.870.11243.144.345.646.848.149.350.651.853.154.355.656.858.159.460.762.063.264.565.867.168.469.771.072.41344.946.147.448.649.951.252.553.755.056.357.658.960.261.562.864.165.466.768.069.370.772.073.374.71446.748.049.350.551.853.154.455.757.058.359.660.962.363.664.966.367.668.970.371.673.074.475.777.11548.649.951.252.553.855.156.457.859.160.461.863.164.565.867.268.569.971.372.674.075.476.878.279.61650.551.853.254.555.857.258.559.961.262.664.065.366.768.169.570.972.373.775.176.577.979.380.882.21752.553.955.256.658.059.360.762.163.564.966.367.769.170.571.973.374.876.277.679.180.582.083.584.91854.656.057.458.860.261.663.064.465.867.268.670.171.573.074.475.977.378.880.381.883.384.886.387.81956.858.259.661.062.463.965.366.868.269.771.172.674.175.577.078.580.081.583.184.686.187.689.290.72059.060.461.963.364.866.367.769.270.772.273.775.276.778.279.881.382.884.485.987.589.190.792.293.82161.362.864.365.867.368.870.371.873.374.976.477.979.581.182.684.285.887.489.090.692.293.895.497.12263.865.366.868.369.871.472.974.576.177.679.280.882.484.085.687.288.990.592.193.895.597.198.8100.52366.367.869.471.072.574.175.777.378.980.582.283.885.487.188.790.492.193.895.597.298.9100.6102.4104.12468.970.572.173.775.377.078.680.381.983.685.286.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92571.773.375.076.678.380.081.783.385.186.888.590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.0111.92674.676.378.079.781.483.184.886.688.490.191.993.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22777.679.481.182.984.686.488.290.091.993.795.597.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.72880.882.684.486.388.189.991.893.795.697.599.4101.3103.3105.2107.2109.2111.2113.2115.2117.3119.3121.4123.5125.62984.286.187.989.891.793.795.697.599.5101.5103.5105.5107.5109.5111.6113.7115.7117.8120.0122.1124.2126.4128.6130.83087.889.791.793.695.697.699.6101.6103.7105.7107.8109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.43191.693.695.697.799.7101.8103.9106.0108.2110.3112.5114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.53295.797.899.9102.0104.2106.3108.5110.7113.0115.2117.5119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.9125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.8131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.4145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.3153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.2162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.4173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.1185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.8199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.4216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.8238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.1265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.3302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.3355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.8436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.3574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4866.4920.8980.41046.21119.91203.31299.71413.61553.11732.91986.32419.6>2419.6

# IDEXX Quanti-Tray/2000

Version Italiano

Quanti-Tray e Defined Substrate Technology sono marchi di proprietà di, e/o registrati da, IDEXX Laboratories, Inc.

e protetti negli Stati Uniti e/o in altri paesi. Protetto dai brevetti degli U.S.A. nn. 5,518,892; 5,620,895; and 5,753,456.

Altre domande di brevetto pendenti. 

# Introduzione

Quanti-Tray/2000 IDEXX sono concepiti per fornire la densità batterica (quantitativa) di campioni da 100 mL utilizzando reagenti Defined

Substrate Technology IDEXX. Aggiungere la miscela di reagente/campione ad un Quanti-Tray/2000, inserire quest’ultimo nel Quanti-Tray

Sealer ed incubare in conformità alle istruzioni fornite con il reagente. Contare quindi il numero di pozzetti positivi e, con l’ausilio della

tabella MPN allegata, determinare il numero più probabile (MPN).



# Contenuto

Questa confezione contiene Quanti-Tray/2000 sterili.



# Supporto Tecnico IDEXX

**00800-4339-9111**

**www.idexx.it/acqua** 

# Istruzioni per l’uso

1. Con una mano, tenere il Quanti-Tray

in posizione verticale, con il lato dei

pozzetti rivolto verso il palmo. 2. Comprimere la parte superiore del

Quanti-Tray in modo da fletterlo verso

il palmo. 3. Aprire il Quanti-Tray tirando la linguetta

in alluminio in direzione opposta al lato

dei pozzetti. Evitare di toccare la parte

interna del foglio protettivo in alluminio

o del dispositivo a pozzetti.

4. Versare direttamente la miscela di

reagente/campione nel Quanti-Tray,

evitando di toccare la linguetta in

alluminio. Dare 2–3 leggeri colpi ai

pozzetti piccoli per eliminare eventuali

bolle d'aria. Consentire alla schiuma di

sedimentare. 5. Una volta versata la miscela, collocare

il Quanti-Tray nel portapozzetti in

gomma del Quanti-Tray Sealer, con

il lto dei pozzetti (in plastica) rivolto

verso il basso, in modo da assicurarne

il corretto posizionamento. 6. Sigillare secondo le istruzioni del

Quanti-Tray Sealer.

7. Incubare secondo le istruzioni fornite

con il reagente.

8. Contare i pozzetti positivi e consultare

la tabella MPN sul retro del foglio delle

istruzioni, per individuare il numero più

probabile (MPN) corretto.

9. Eliminare el terreno in accordo alle

Buone pratiche di laboratorio.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA**IDEXX Quanti-Tray/2000 Tabella MPN # Pozzetti** **Grande Positivi** **# Pozzetti Piccoli Positivi 0** **1 2** **3 4** **5 6** **7 8** **9 10** **11 12** **13 14** **15 16** **17 18** **19 20** **21 22** **23 24** 0<11.02.03.04.05.06.07.08.09.010.011.012.013.014.115.116.117.118.119.120.221.222.223.324.311.02.03.04.05.06.07.18.19.110.111.112.113.214.215.216.217.318.319.320.421.422.423.524.525.622.03.04.15.16.17.18.19.210.211.212.213.314.315.416.417.418.519.520.621.622.723.724.825.826.933.14.15.16.17.28.29.210.311.312.413.414.515.516.517.618.619.720.821.822.923.925.026.127.128.244.15.26.27.28.39.310.411.412.513.514.615.616.717.818.819.921.022.023.124.225.326.327.428.529.655.26.37.38.49.410.511.512.613.714.715.816.917.919.020.121.222.223.324.425.526.627.728.829.931.066.37.48.49.510.611.612.713.814.916.017.018.119.220.321.422.523.624.725.826.928.029.130.231.332.477.58.59.610.711.812.813.915.016.117.218.319.420.521.622.723.824.926.027.128.329.430.531.632.833.988.69.710.811.913.014.115.216.317.418.519.620.721.822.924.125.226.327.428.629.730.832.033.134.335.499.810.912.013.114.215.316.417.618.719.820.922.023.224.325.426.627.728.930.031.232.333.534.635.837.01011.012.113.214.415.516.617.718.920.021.122.323.424.625.726.928.029.230.331.532.733.835.036.237.438.61112.213.414.515.616.817.919.120.221.422.523.724.826.027.228.329.530.731.933.034.235.436.637.839.040.21213.514.615.816.918.119.320.421.622.823.925.126.327.528.629.831.032.233.434.635.837.038.239.540.741.91314.816.017.118.319.520.621.823.024.225.426.627.829.030.231.432.633.835.036.237.538.739.941.242.443.61416.117.318.519.720.922.123.324.525.726.928.129.330.531.733.034.235.436.737.939.140.441.642.944.245.41517.518.719.921.122.323.524.725.927.228.429.630.932.133.334.635.837.138.439.640.942.243.444.746.047.31618.920.121.322.623.825.026.227.528.730.031.232.533.735.036.337.538.840.141.442.744.045.346.647.949.21720.321.622.824.125.326.627.829.130.331.632.934.135.436.738.039.340.641.943.244.545.947.248.549.851.21821.823.124.325.626.928.129.430.732.033.334.635.937.238.539.841.142.443.845.146.547.849.250.551.953.21923.324.625.927.228.529.831.132.433.735.036.337.639.040.341.643.044.345.747.148.449.851.252.654.055.42024.926.227.528.830.131.532.834.135.436.838.139.540.842.243.644.946.347.749.150.551.953.354.756.157.62126.527.929.230.531.833.234.535.937.338.640.041.442.844.145.546.948.449.851.252.654.155.556.958.459.92228.229.530.932.333.635.036.437.739.140.541.943.344.846.247.649.050.551.953.454.856.357.859.360.862.32329.931.332.734.135.536.838.339.741.142.543.945.446.848.349.751.252.754.255.657.158.660.261.763.264.72431.733.134.535.937.338.840.241.743.144.646.047.549.050.552.053.555.056.558.059.561.162.664.265.867.32533.635.036.437.939.340.842.243.745.246.748.249.751.252.754.355.857.358.960.562.063.665.266.868.470.02635.536.938.439.941.442.844.345.947.448.950.452.053.555.156.758.259.861.463.064.766.367.969.671.272.92737.438.940.442.043.545.046.548.149.651.252.854.456.057.659.260.862.464.165.767.469.170.872.574.275.92839.541.042.644.145.747.348.850.452.053.655.256.958.560.261.863.565.266.968.670.372.073.775.577.379.02941.743.244.846.448.049.651.252.854.556.157.859.561.262.964.666.368.069.871.573.375.176.978.780.582.43043.945.547.148.750.452.053.755.457.158.860.562.264.065.767.569.371.072.974.776.578.380.282.184.085.93146.247.949.551.252.954.656.358.159.861.663.365.166.968.770.572.474.276.178.079.981.883.785.787.689.63248.750.452.153.855.657.359.160.962.764.566.368.270.071.973.875.777.679.581.583.585.487.589.591.593.63351.253.054.856.558.360.262.063.865.767.669.571.473.375.277.279.281.283.285.287.389.391.493.695.797.83453.955.757.659.461.363.165.067.068.970.872.874.876.878.880.882.985.087.189.291.493.595.797.9100.2102.43556.858.660.562.464.466.368.370.372.374.376.378.480.582.684.786.989.191.393.595.798.0100.3102.6105.0107.33659.861.763.765.767.769.771.773.875.978.080.182.384.586.788.991.293.595.898.1100.5102.9105.3107.7110.2112.73762.965.067.069.171.273.375.477.679.882.084.286.588.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.9118.63866.368.470.672.774.977.179.481.683.986.288.691.093.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.2125.03970.072.274.476.778.981.383.686.088.490.993.495.998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.2132.24073.876.278.580.983.385.788.290.893.395.998.5101.2103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.0140.34178.080.583.085.588.090.693.395.998.7101.4104.3107.1110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.9149.54282.685.287.890.593.296.098.8101.7104.6107.6110.6113.7116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.1160.24387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.1124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.2172.84493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.6133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.9188.24599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.6143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.2207.546106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.5156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.4233.347114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.4172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.5270.048123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.0193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.0328.249135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6435.2Quanti-Tray e Defined Substrate Technology sono marchi di proprietà di, e/o registrati da, IDEXX Laboratories, Inc. e protetti negli Stati Uniti e/o in altri paesi. Protetto dai brevetti degli U.S.A. nn. 4,925,789; 5,429,933; 5,518,892. Altre domande di brevetto pendenti.**IDEXX Quanti-Tray/2000 Tabella MPN # Pozzetti** **Grande Positivi** **# Pozzetti Piccoli Positivi 25** **26 27** **28 29** **30 31** **32 33** **34 35** **36 37** **38 39** **40 41** **42 43** **44 45** **46 47** **48**025.326.427.428.429.530.531.532.633.634.735.736.837.838.940.041.042.143.144.245.346.347.448.549.5126.627.728.729.830.831.932.934.035.036.137.238.239.340.441.442.543.644.745.746.847.949.050.151.2227.929.030.031.132.233.234.335.436.537.538.639.740.841.943.044.045.146.247.348.449.550.651.752.8329.330.431.432.533.634.735.836.837.939.040.141.242.343.444.545.646.747.848.950.051.252.353.454.5430.731.832.833.935.036.137.238.339.440.541.642.843.945.046.147.248.349.550.651.752.954.055.156.3532.133.234.335.436.537.638.739.941.042.143.244.445.546.647.748.950.051.252.353.554.655.856.958.1633.534.735.836.938.039.240.341.442.643.744.846.047.148.349.450.651.752.954.155.256.457.658.759.9735.036.237.338.439.640.741.943.044.245.346.547.748.850.051.252.353.554.755.957.158.359.460.661.8836.637.738.940.041.242.343.544.745.947.048.249.450.651.853.054.155.356.557.759.060.261.462.663.8938.139.340.541.642.844.045.246.447.648.850.051.252.453.654.856.057.258.459.760.962.163.464.665.81039.740.942.143.344.545.746.948.149.350.651.853.054.255.556.757.959.260.461.762.964.265.466.767.91141.442.643.845.046.347.548.749.951.252.453.754.956.157.458.659.961.262.463.765.066.367.568.870.11243.144.345.646.848.149.350.651.853.154.355.656.858.159.460.762.063.264.565.867.168.469.771.072.41344.946.147.448.649.951.252.553.755.056.357.658.960.261.562.864.165.466.768.069.370.772.073.374.71446.748.049.350.551.853.154.455.757.058.359.660.962.363.664.966.367.668.970.371.673.074.475.777.11548.649.951.252.553.855.156.457.859.160.461.863.164.565.867.268.569.971.372.674.075.476.878.279.61650.551.853.254.555.857.258.559.961.262.664.065.366.768.169.570.972.373.775.176.577.979.380.882.21752.553.955.256.658.059.360.762.163.564.966.367.769.170.571.973.374.876.277.679.180.582.083.584.91854.656.057.458.860.261.663.064.465.867.268.670.171.573.074.475.977.378.880.381.883.384.886.387.81956.858.259.661.062.463.965.366.868.269.771.172.674.175.577.078.580.081.583.184.686.187.689.290.72059.060.461.963.364.866.367.769.270.772.273.775.276.778.279.881.382.884.485.987.589.190.792.293.82161.362.864.365.867.368.870.371.873.374.976.477.979.581.182.684.285.887.489.090.692.293.895.497.12263.865.366.868.369.871.472.974.576.177.679.280.882.484.085.687.288.990.592.193.895.597.198.8100.52366.367.869.471.072.574.175.777.378.980.582.283.885.487.188.790.492.193.895.597.298.9100.6102.4104.12468.970.572.173.775.377.078.680.381.983.685.286.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92571.773.375.076.678.380.081.783.385.186.888.590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.0111.92674.676.378.079.781.483.184.886.688.490.191.993.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22777.679.481.182.984.686.488.290.091.993.795.597.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.72880.882.684.486.388.189.991.893.795.697.599.4101.3103.3105.2107.2109.2111.2113.2115.2117.3119.3121.4123.5125.62984.286.187.989.891.793.795.697.599.5101.5103.5105.5107.5109.5111.6113.7115.7117.8120.0122.1124.2126.4128.6130.83087.889.791.793.695.697.699.6101.6103.7105.7107.8109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.43191.693.695.697.799.7101.8103.9106.0108.2110.3112.5114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.53295.797.899.9102.0104.2106.3108.5110.7113.0115.2117.5119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.9125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.8131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.4145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.3153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.2162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.4173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.1185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.8199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.4216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.8238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.1265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.3302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.3355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.8436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.3574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4866.4920.8980.41046.21119.91203.31299.71413.61553.11732.91986.32419.6>2419.6

# IDEXX Quanti-Tray/2000

Versíon Español

Quanti-Tray y Defined Substrate Technology son marcas o marcas registradas de IDEXX Laboratories, Inc. en los

Estados Unidos y/o otros paises. Patente norteamericanas numero 5,518,892; 5,620,895; and 5,753,456. Otras patentes

pendientes de concesión. 

# Introducción

Los dispositivos Quanti-Tray/2000 de IDEXX están diseñados para producir recuentos bacterianos cuantificados de muestras de 100 mL,

al ser utilizadas con productos de reactivos de la Tecnología del Sustrato Definido (Defined Substrate Technology) de IDEXX. Agregue

la mezcla de reactivo y muestra a un dispositivo Quanti-Tray/2000, séllelo en el Quanti-Tray Sealer (Sellador) e incúbela según las

instrucciones del reactivo. Luego cuente el número de celdas positivas y utilice la tabla de NMP adjunta para determinar el Número Más

Probable (NMP).



# Contenido

Este paquete contiene dispositivos Quanti-Tray/2000 estériles.



# Soporte técnico de IDEXX

**00800-4339-9111**

**www.idexx.es/agua** 

# Instrucciones para el usuario

1. Sostenga en una mano el dispositivo

Quanti-Tray en posición vertical, con

el lado de las celdas orientado hacia

la palma. 2. Apriete la parte superior de el

dispositivo Quanti-Tray de modo de

doblarla hacia la palma. 3. Abra el dispositivo Quanti-Tray

desprendiendo la lengüeta metálica del

lado que contiene las celdas. Evite tocar

el interior del metal o del dispositivo.

4. Vierta la mezcla de reactivo y la

muestra directamente dentro de el

dispositivo Quanti-Tray, evitando tocar

la lengüeta metálica. Golpear los

pequeños pocillos 2 o 3 veces para

eliminar posibles burbujas de aire.

Deja reposar la espuma. 5. Coloque el dispositivo Quanti-Tray lleno

de muestra sobre el portadispositivo

de goma del sellador Quanti-Tray,

orientando el lado de las celdas

(plásticas) del dispositivo hacia

abajo de manera que quepa en el

portadispositivo. 6. Selle el dispositivo según las

instrucciones del sellador.

7. Incube de acuerdo con las

instrucciones del reactivo.

8. Cuente las celdas positivas. Para

determinar el número más probable,

recurra a la tabla NMP al dorso de

esta hoja de instrucciones.

9. Verter el medio de cultivo conforme a

Buenas Prácticas del Laboratorio.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA**IDEXX Quanti-Tray/2000 Tabla, número más probable # Pocillos** **Grandes Positivos** **# Pocillos pequeños positivos 0** **1 2** **3 4** **5 6** **7 8** **9 10** **11 12** **13 14** **15 16** **17 18** **19 20** **21 22** **23 24** 0<11.02.03.04.05.06.07.08.09.010.011.012.013.014.115.116.117.118.119.120.221.222.223.324.311.02.03.04.05.06.07.18.19.110.111.112.113.214.215.216.217.318.319.320.421.422.423.524.525.622.03.04.15.16.17.18.19.210.211.212.213.314.315.416.417.418.519.520.621.622.723.724.825.826.933.14.15.16.17.28.29.210.311.312.413.414.515.516.517.618.619.720.821.822.923.925.026.127.128.244.15.26.27.28.39.310.411.412.513.514.615.616.717.818.819.921.022.023.124.225.326.327.428.529.655.26.37.38.49.410.511.512.613.714.715.816.917.919.020.121.222.223.324.425.526.627.728.829.931.066.37.48.49.510.611.612.713.814.916.017.018.119.220.321.422.523.624.725.826.928.029.130.231.332.477.58.59.610.711.812.813.915.016.117.218.319.420.521.622.723.824.926.027.128.329.430.531.632.833.988.69.710.811.913.014.115.216.317.418.519.620.721.822.924.125.226.327.428.629.730.832.033.134.335.499.810.912.013.114.215.316.417.618.719.820.922.023.224.325.426.627.728.930.031.232.333.534.635.837.01011.012.113.214.415.516.617.718.920.021.122.323.424.625.726.928.029.230.331.532.733.835.036.237.438.61112.213.414.515.616.817.919.120.221.422.523.724.826.027.228.329.530.731.933.034.235.436.637.839.040.21213.514.615.816.918.119.320.421.622.823.925.126.327.528.629.831.032.233.434.635.837.038.239.540.741.91314.816.017.118.319.520.621.823.024.225.426.627.829.030.231.432.633.835.036.237.538.739.941.242.443.61416.117.318.519.720.922.123.324.525.726.928.129.330.531.733.034.235.436.737.939.140.441.642.944.245.41517.518.719.921.122.323.524.725.927.228.429.630.932.133.334.635.837.138.439.640.942.243.444.746.047.31618.920.121.322.623.825.026.227.528.730.031.232.533.735.036.337.538.840.141.442.744.045.346.647.949.21720.321.622.824.125.326.627.829.130.331.632.934.135.436.738.039.340.641.943.244.545.947.248.549.851.21821.823.124.325.626.928.129.430.732.033.334.635.937.238.539.841.142.443.845.146.547.849.250.551.953.21923.324.625.927.228.529.831.132.433.735.036.337.639.040.341.643.044.345.747.148.449.851.252.654.055.42024.926.227.528.830.131.532.834.135.436.838.139.540.842.243.644.946.347.749.150.551.953.354.756.157.62126.527.929.230.531.833.234.535.937.338.640.041.442.844.145.546.948.449.851.252.654.155.556.958.459.92228.229.530.932.333.635.036.437.739.140.541.943.344.846.247.649.050.551.953.454.856.357.859.360.862.32329.931.332.734.135.536.838.339.741.142.543.945.446.848.349.751.252.754.255.657.158.660.261.763.264.72431.733.134.535.937.338.840.241.743.144.646.047.549.050.552.053.555.056.558.059.561.162.664.265.867.32533.635.036.437.939.340.842.243.745.246.748.249.751.252.754.355.857.358.960.562.063.665.266.868.470.02635.536.938.439.941.442.844.345.947.448.950.452.053.555.156.758.259.861.463.064.766.367.969.671.272.92737.438.940.442.043.545.046.548.149.651.252.854.456.057.659.260.862.464.165.767.469.170.872.574.275.92839.541.042.644.145.747.348.850.452.053.655.256.958.560.261.863.565.266.968.670.372.073.775.577.379.02941.743.244.846.448.049.651.252.854.556.157.859.561.262.964.666.368.069.871.573.375.176.978.780.582.43043.945.547.148.750.452.053.755.457.158.860.562.264.065.767.569.371.072.974.776.578.380.282.184.085.93146.247.949.551.252.954.656.358.159.861.663.365.166.968.770.572.474.276.178.079.981.883.785.787.689.63248.750.452.153.855.657.359.160.962.764.566.368.270.071.973.875.777.679.581.583.585.487.589.591.593.63351.253.054.856.558.360.262.063.865.767.669.571.473.375.277.279.281.283.285.287.389.391.493.695.797.83453.955.757.659.461.363.165.067.068.970.872.874.876.878.880.882.985.087.189.291.493.595.797.9100.2102.43556.858.660.562.464.466.368.370.372.374.376.378.480.582.684.786.989.191.393.595.798.0100.3102.6105.0107.33659.861.763.765.767.769.771.773.875.978.080.182.384.586.788.991.293.595.898.1100.5102.9105.3107.7110.2112.73762.965.067.069.171.273.375.477.679.882.084.286.588.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.9118.63866.368.470.672.774.977.179.481.683.986.288.691.093.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.2125.03970.072.274.476.778.981.383.686.088.490.993.495.998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.2132.24073.876.278.580.983.385.788.290.893.395.998.5101.2103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.0140.34178.080.583.085.588.090.693.395.998.7101.4104.3107.1110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.9149.54282.685.287.890.593.296.098.8101.7104.6107.6110.6113.7116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.1160.24387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.1124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.2172.84493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.6133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.9188.24599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.6143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.2207.546106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.5156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.4233.347114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.4172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.5270.048123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.0193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.0328.249135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6435.2Quanti-Tray y Defined Substrate Technology son marcas o marcas registradas de IDEXX Laboratories, Inc. en los Estados Unidos y/o otros paises. Patente norteamericanas numero 4,925,789; 5,429,933; 5,518,892. Otras patentes pendientes de concesión.**IDEXX Quanti-Tray/2000 Tabla, número más probable # Pocillos** **Grandes Positivos** **# Pocillos pequeños positivos 25** **26 27** **28 29** **30 31** **32 33** **34 35** **36 37** **38 39** **40 41** **42 43** **44 45** **46 47** **48**025.326.427.428.429.530.531.532.633.634.735.736.837.838.940.041.042.143.144.245.346.347.448.549.5126.627.728.729.830.831.932.934.035.036.137.238.239.340.441.442.543.644.745.746.847.949.050.151.2227.929.030.031.132.233.234.335.436.537.538.639.740.841.943.044.045.146.247.348.449.550.651.752.8329.330.431.432.533.634.735.836.837.939.040.141.242.343.444.545.646.747.848.950.051.252.353.454.5430.731.832.833.935.036.137.238.339.440.541.642.843.945.046.147.248.349.550.651.752.954.055.156.3532.133.234.335.436.537.638.739.941.042.143.244.445.546.647.748.950.051.252.353.554.655.856.958.1633.534.735.836.938.039.240.341.442.643.744.846.047.148.349.450.651.752.954.155.256.457.658.759.9735.036.237.338.439.640.741.943.044.245.346.547.748.850.051.252.353.554.755.957.158.359.460.661.8836.637.738.940.041.242.343.544.745.947.048.249.450.651.853.054.155.356.557.759.060.261.462.663.8938.139.340.541.642.844.045.246.447.648.850.051.252.453.654.856.057.258.459.760.962.163.464.665.81039.740.942.143.344.545.746.948.149.350.651.853.054.255.556.757.959.260.461.762.964.265.466.767.91141.442.643.845.046.347.548.749.951.252.453.754.956.157.458.659.961.262.463.765.066.367.568.870.11243.144.345.646.848.149.350.651.853.154.355.656.858.159.460.762.063.264.565.867.168.469.771.072.41344.946.147.448.649.951.252.553.755.056.357.658.960.261.562.864.165.466.768.069.370.772.073.374.71446.748.049.350.551.853.154.455.757.058.359.660.962.363.664.966.367.668.970.371.673.074.475.777.11548.649.951.252.553.855.156.457.859.160.461.863.164.565.867.268.569.971.372.674.075.476.878.279.61650.551.853.254.555.857.258.559.961.262.664.065.366.768.169.570.972.373.775.176.577.979.380.882.21752.553.955.256.658.059.360.762.163.564.966.367.769.170.571.973.374.876.277.679.180.582.083.584.91854.656.057.458.860.261.663.064.465.867.268.670.171.573.074.475.977.378.880.381.883.384.886.387.81956.858.259.661.062.463.965.366.868.269.771.172.674.175.577.078.580.081.583.184.686.187.689.290.72059.060.461.963.364.866.367.769.270.772.273.775.276.778.279.881.382.884.485.987.589.190.792.293.82161.362.864.365.867.368.870.371.873.374.976.477.979.581.182.684.285.887.489.090.692.293.895.497.12263.865.366.868.369.871.472.974.576.177.679.280.882.484.085.687.288.990.592.193.895.597.198.8100.52366.367.869.471.072.574.175.777.378.980.582.283.885.487.188.790.492.193.895.597.298.9100.6102.4104.12468.970.572.173.775.377.078.680.381.983.685.286.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92571.773.375.076.678.380.081.783.385.186.888.590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.0111.92674.676.378.079.781.483.184.886.688.490.191.993.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22777.679.481.182.984.686.488.290.091.993.795.597.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.72880.882.684.486.388.189.991.893.795.697.599.4101.3103.3105.2107.2109.2111.2113.2115.2117.3119.3121.4123.5125.62984.286.187.989.891.793.795.697.599.5101.5103.5105.5107.5109.5111.6113.7115.7117.8120.0122.1124.2126.4128.6130.83087.889.791.793.695.697.699.6101.6103.7105.7107.8109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.43191.693.695.697.799.7101.8103.9106.0108.2110.3112.5114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.53295.797.899.9102.0104.2106.3108.5110.7113.0115.2117.5119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.9125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.8131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.4145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.3153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.2162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.4173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.1185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.8199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.4216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.8238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.1265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.3302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.3355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.8436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.3574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4866.4920.8980.41046.21119.91203.31299.71413.61553.11732.91986.32419.6>2419.6

# IDEXX Quanti-Tray/2000

Deutsche Version

Quanti-Tray und Defined Substrate Technology sind Schutzmarken oder eingetragene Schutzmarken von

IDEXX Laboratories, Inc. in den Vereinigten Staaten und/oder in anderen Ländern. Geschützt durch U.S.-Patent

Nummern 5,518,892; 5,620,895; and 5,753,456. Andere Patenten angemeldet. 

# Einführung

Quanti-Tray/2000 von IDEXX dienen zur quantitativen Bakterienzählung in 100-mL-Proben unter Verwendung von IDEXX Defined

Substrate Technology-Reagenzien. Die Mischung aus Probe und Reagenz in einen Quanti-Tray/2000 geben, diesen in einem Quanti-TrayVersiegelungsgerät verschließen und laut Reagenz-Packungsbeilage inkubieren. Anschließend die Anzahl der positiven Vertiefungen

zählen und mit der beigefügten MPN-Tabelle die “Wahrscheinlichste Anzahl” (MPN) bestimmen.



# Inhalt

Diese Packung enthält sterile Quanti-Tray/2000.



# IDEXX Technischer Dienst

**00800-4339-9111**

**www.idexx.de/wasser** 

# Gebrauchsanweisung

1. Quanti-Tray mit einer Hand fassen und

senkrecht halten, wobei die Seite mit

den Vertiefungen zur Handfläche zeigt. 2. Den oberen Teil des Quanti-Trays

zusammendrücken, so daß sich der

Träger zur Handfläche hin biegt. 3. Quanti-Tray öffnen, indem die

Folienlasche von der Seite mit den

Vertiefungen abgezogen wird. Die

Innenseite der Folie oder des Trays

nicht berühren.

4. Die Mischung aus Probe und

Reagenz direkt in einen Quanti-Tray

gießen. Dabei eine Berührung der

Folienlasche vermeiden. Seitlich

2–3 mal an die kleinen Vertiefungen

klopfen, um eventuelle Luftblasen zu

entfernen. Schaum einwirken lassen. 5. Den mit der Probe gefüllten QuantiTray auf die Gummi-Trägerunterlage

des Quanti-Tray-Versiegelungsgeräts

legen. Die Seite mit den Vertiefungen

(Kunststoff) des Quanti-Trays muß

dabei nach unten zeigen. 6. Das Verschließen erfolgt

laut Anweisungen für das

Versiegelungsgerät.

7. Entsprechend den

Reagenzanweisungen inkubieren.

8. Die positiven Vertiefungen zählen

und mit Hilfe der MPN-Tabelle auf der

Rückseite dieser Packungsbeilage

die Wahrscheinlichste Anzahl (MPN)

ermitteln.

9. Entsorgen Sie die Quanti-Trays gemäß

guter Laborpraxis.

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA**IDEXX Quanti-Tray/2000 MPN-Tabelle # Positiven** **Großen Vertiefungen** **# Positiven Kleinen Vertiefungen 0** **1 2** **3 4** **5 6** **7 8** **9 10** **11 12** **13 14** **15 16** **17 18** **19 20** **21 22** **23 24** 0<11.02.03.04.05.06.07.08.09.010.011.012.013.014.115.116.117.118.119.120.221.222.223.324.311.02.03.04.05.06.07.18.19.110.111.112.113.214.215.216.217.318.319.320.421.422.423.524.525.622.03.04.15.16.17.18.19.210.211.212.213.314.315.416.417.418.519.520.621.622.723.724.825.826.933.14.15.16.17.28.29.210.311.312.413.414.515.516.517.618.619.720.821.822.923.925.026.127.128.244.15.26.27.28.39.310.411.412.513.514.615.616.717.818.819.921.022.023.124.225.326.327.428.529.655.26.37.38.49.410.511.512.613.714.715.816.917.919.020.121.222.223.324.425.526.627.728.829.931.066.37.48.49.510.611.612.713.814.916.017.018.119.220.321.422.523.624.725.826.928.029.130.231.332.477.58.59.610.711.812.813.915.016.117.218.319.420.521.622.723.824.926.027.128.329.430.531.632.833.988.69.710.811.913.014.115.216.317.418.519.620.721.822.924.125.226.327.428.629.730.832.033.134.335.499.810.912.013.114.215.316.417.618.719.820.922.023.224.325.426.627.728.930.031.232.333.534.635.837.01011.012.113.214.415.516.617.718.920.021.122.323.424.625.726.928.029.230.331.532.733.835.036.237.438.61112.213.414.515.616.817.919.120.221.422.523.724.826.027.228.329.530.731.933.034.235.436.637.839.040.21213.514.615.816.918.119.320.421.622.823.925.126.327.528.629.831.032.233.434.635.837.038.239.540.741.91314.816.017.118.319.520.621.823.024.225.426.627.829.030.231.432.633.835.036.237.538.739.941.242.443.61416.117.318.519.720.922.123.324.525.726.928.129.330.531.733.034.235.436.737.939.140.441.642.944.245.41517.518.719.921.122.323.524.725.927.228.429.630.932.133.334.635.837.138.439.640.942.243.444.746.047.31618.920.121.322.623.825.026.227.528.730.031.232.533.735.036.337.538.840.141.442.744.045.346.647.949.21720.321.622.824.125.326.627.829.130.331.632.934.135.436.738.039.340.641.943.244.545.947.248.549.851.21821.823.124.325.626.928.129.430.732.033.334.635.937.238.539.841.142.443.845.146.547.849.250.551.953.21923.324.625.927.228.529.831.132.433.735.036.337.639.040.341.643.044.345.747.148.449.851.252.654.055.42024.926.227.528.830.131.532.834.135.436.838.139.540.842.243.644.946.347.749.150.551.953.354.756.157.62126.527.929.230.531.833.234.535.937.338.640.041.442.844.145.546.948.449.851.252.654.155.556.958.459.92228.229.530.932.333.635.036.437.739.140.541.943.344.846.247.649.050.551.953.454.856.357.859.360.862.32329.931.332.734.135.536.838.339.741.142.543.945.446.848.349.751.252.754.255.657.158.660.261.763.264.72431.733.134.535.937.338.840.241.743.144.646.047.549.050.552.053.555.056.558.059.561.162.664.265.867.32533.635.036.437.939.340.842.243.745.246.748.249.751.252.754.355.857.358.960.562.063.665.266.868.470.02635.536.938.439.941.442.844.345.947.448.950.452.053.555.156.758.259.861.463.064.766.367.969.671.272.92737.438.940.442.043.545.046.548.149.651.252.854.456.057.659.260.862.464.165.767.469.170.872.574.275.92839.541.042.644.145.747.348.850.452.053.655.256.958.560.261.863.565.266.968.670.372.073.775.577.379.02941.743.244.846.448.049.651.252.854.556.157.859.561.262.964.666.368.069.871.573.375.176.978.780.582.43043.945.547.148.750.452.053.755.457.158.860.562.264.065.767.569.371.072.974.776.578.380.282.184.085.93146.247.949.551.252.954.656.358.159.861.663.365.166.968.770.572.474.276.178.079.981.883.785.787.689.63248.750.452.153.855.657.359.160.962.764.566.368.270.071.973.875.777.679.581.583.585.487.589.591.593.63351.253.054.856.558.360.262.063.865.767.669.571.473.375.277.279.281.283.285.287.389.391.493.695.797.83453.955.757.659.461.363.165.067.068.970.872.874.876.878.880.882.985.087.189.291.493.595.797.9100.2102.43556.858.660.562.464.466.368.370.372.374.376.378.480.582.684.786.989.191.393.595.798.0100.3102.6105.0107.33659.861.763.765.767.769.771.773.875.978.080.182.384.586.788.991.293.595.898.1100.5102.9105.3107.7110.2112.73762.965.067.069.171.273.375.477.679.882.084.286.588.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.9118.63866.368.470.672.774.977.179.481.683.986.288.691.093.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.2125.03970.072.274.476.778.981.383.686.088.490.993.495.998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.2132.24073.876.278.580.983.385.788.290.893.395.998.5101.2103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.0140.34178.080.583.085.588.090.693.395.998.7101.4104.3107.1110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.9149.54282.685.287.890.593.296.098.8101.7104.6107.6110.6113.7116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.1160.24387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.1124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.2172.84493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.6133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.9188.24599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.6143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.2207.546106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.5156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.4233.347114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.4172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.5270.048123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.0193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.0328.249135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6435.2Quanti-Tray und Defined Substrate Technology sind Schutzmarken oder eingetragene Schutzmarken von IDEXX Laboratories, Inc. in den Vereinigten Staaten und/oder in anderen Ländern. Geschützt durch U.S.-Patent Nummern 4,925,789; 5,429,933; 5,518,892. Andere Patenten angemeldet.**IDEXX Quanti-Tray/2000 MPN-Tabelle # Positiven** **Großen Vertiefungen** **# Positiven Kleinen Vertiefungen 25** **26 27** **28 29** **30 31** **32 33** **34 35** **36 37** **38 39** **40 41** **42 43** **44 45** **46 47** **48**025.326.427.428.429.530.531.532.633.634.735.736.837.838.940.041.042.143.144.245.346.347.448.549.5126.627.728.729.830.831.932.934.035.036.137.238.239.340.441.442.543.644.745.746.847.949.050.151.2227.929.030.031.132.233.234.335.436.537.538.639.740.841.943.044.045.146.247.348.449.550.651.752.8329.330.431.432.533.634.735.836.837.939.040.141.242.343.444.545.646.747.848.950.051.252.353.454.5430.731.832.833.935.036.137.238.339.440.541.642.843.945.046.147.248.349.550.651.752.954.055.156.3532.133.234.335.436.537.638.739.941.042.143.244.445.546.647.748.950.051.252.353.554.655.856.958.1633.534.735.836.938.039.240.341.442.643.744.846.047.148.349.450.651.752.954.155.256.457.658.759.9735.036.237.338.439.640.741.943.044.245.346.547.748.850.051.252.353.554.755.957.158.359.460.661.8836.637.738.940.041.242.343.544.745.947.048.249.450.651.853.054.155.356.557.759.060.261.462.663.8938.139.340.541.642.844.045.246.447.648.850.051.252.453.654.856.057.258.459.760.962.163.464.665.81039.740.942.143.344.545.746.948.149.350.651.853.054.255.556.757.959.260.461.762.964.265.466.767.91141.442.643.845.046.347.548.749.951.252.453.754.956.157.458.659.961.262.463.765.066.367.568.870.11243.144.345.646.848.149.350.651.853.154.355.656.858.159.460.762.063.264.565.867.168.469.771.072.41344.946.147.448.649.951.252.553.755.056.357.658.960.261.562.864.165.466.768.069.370.772.073.374.71446.748.049.350.551.853.154.455.757.058.359.660.962.363.664.966.367.668.970.371.673.074.475.777.11548.649.951.252.553.855.156.457.859.160.461.863.164.565.867.268.569.971.372.674.075.476.878.279.61650.551.853.254.555.857.258.559.961.262.664.065.366.768.169.570.972.373.775.176.577.979.380.882.21752.553.955.256.658.059.360.762.163.564.966.367.769.170.571.973.374.876.277.679.180.582.083.584.91854.656.057.458.860.261.663.064.465.867.268.670.171.573.074.475.977.378.880.381.883.384.886.387.81956.858.259.661.062.463.965.366.868.269.771.172.674.175.577.078.580.081.583.184.686.187.689.290.72059.060.461.963.364.866.367.769.270.772.273.775.276.778.279.881.382.884.485.987.589.190.792.293.82161.362.864.365.867.368.870.371.873.374.976.477.979.581.182.684.285.887.489.090.692.293.895.497.12263.865.366.868.369.871.472.974.576.177.679.280.882.484.085.687.288.990.592.193.895.597.198.8100.52366.367.869.471.072.574.175.777.378.980.582.283.885.487.188.790.492.193.895.597.298.9100.6102.4104.12468.970.572.173.775.377.078.680.381.983.685.286.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92571.773.375.076.678.380.081.783.385.186.888.590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.0111.92674.676.378.079.781.483.184.886.688.490.191.993.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22777.679.481.182.984.686.488.290.091.993.795.597.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.72880.882.684.486.388.189.991.893.795.697.599.4101.3103.3105.2107.2109.2111.2113.2115.2117.3119.3121.4123.5125.62984.286.187.989.891.793.795.697.599.5101.5103.5105.5107.5109.5111.6113.7115.7117.8120.0122.1124.2126.4128.6130.83087.889.791.793.695.697.699.6101.6103.7105.7107.8109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.43191.693.695.697.799.7101.8103.9106.0108.2110.3112.5114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.53295.797.899.9102.0104.2106.3108.5110.7113.0115.2117.5119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.9125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.8131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.4145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.3153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.2162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.4173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.1185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.8199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.4216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.8238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.1265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.3302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.3355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.8436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.3574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4866.4920.8980.41046.21119.91203.31299.71413.61553.11732.91986.32419.6>2419.6

# IDEXX Quanti-Tray/2000

日本語版

Quanti-Tray および Defined Substrate Technology は、 米国および / または他国の IDEXX Laboratories,

Inc, の商標または登録商標です。 米国特許番号 5,518,892; 5,620,895; and 5,753,456 によって保護されています。他の特許は出願中です。 

# はじめに

IDEXX Quanti-Tray/2000 は、 IDEXX が特許を取得している Defined Substrate Technology (特定酵素基質法)に基づいた Colilert を用い、 検

体 100 mL 中の最確数を測定するシステムです。 Colilert と検体の混合液を Quanti-Tray/2000 に加え、 シーラーで封入し、取扱説明書に従って培

養してください。 その後、陽性ウェル数を数え、添付の最確数 (MPN) 表を用いて、 MPN を判定します。



# 内容

このパッケージには、滅菌 Quanti-Tray/2000 が 100 個入っています。



# テクニカル・サポート

アイデックス ラボラトリーズ株式会社

TEL:0422-71-5921

**www.idexx.co.jp/water** 

# 操作手順

1. ウェル側を手のひらに向けて、 トレイ を

垂直に持ってください。 2. トレイ が手のひらに向かって曲がるよう

に、 トレイ の上部を強く握ってくださ

い。 3. トレイのつまみをウェル側から引き離し

て、注入口を広げて下さい。 トレイのつ

まみまたはトレイの内部に触れないように

してください。

4. トレイのつまみにかからないように直接、

Colilert と検体の混合液を注いでくださ

い。 小さいウェルを 2 ~ 3 回軽くたた

き、気泡を抜いてください。 5. 検体の入った トレイ を シーラーの ゴム

製シートの上に、 ウェル側 を下に向けて

セットしてください。 6. シーラーの 取扱説明書に従って、密封して

ください。

7. Colilert の取扱説明書に従って、培養して

ください。

8. 大小の陽性ウェル数を数え、 添付の最

確数( MPN) 表を使用して MPN を求めて

ください。

9. 検体及びトレイの廃棄は高圧滅菌してく

ださい。

IDEXX Laboratories, Inc.

One IDEXX Drive

Westbrook, Maine 04092 USA**IDEXX Quanti-Tray/2000 MPN**表大きいウェル陽性数小さいウェル陽性数**0 1** **2 3** **4 5** **6 7** **8 9** **10 11** **12 13** **14 15** **16 17** **18 19** **20 21** **22 23** **24**0<11.02.03.04.05.06.07.08.09.010.011.012.013.014.115.116.117.118.119.120.221.222.223.324.311.02.03.04.05.06.07.18.19.110.111.112.113.214.215.216.217.318.319.320.421.422.423.524.525.622.03.04.15.16.17.18.19.210.211.212.213.314.315.416.417.418.519.520.621.622.723.724.825.826.933.14.15.16.17.28.29.210.311.312.413.414.515.516.517.618.619.720.821.822.923.925.026.127.128.244.15.26.27.28.39.310.411.412.513.514.615.616.717.818.819.921.022.023.124.225.326.327.428.529.655.26.37.38.49.410.511.512.613.714.715.816.917.919.020.121.222.223.324.425.526.627.728.829.931.066.37.48.49.510.611.612.713.814.916.017.018.119.220.321.422.523.624.725.826.928.029.130.231.332.477.58.59.610.711.812.813.915.016.117.218.319.420.521.622.723.824.926.027.128.329.430.531.632.833.988.69.710.811.913.014.115.216.317.418.519.620.721.822.924.125.226.327.428.629.730.832.033.134.335.499.810.912.013.114.215.316.417.618.719.820.922.023.224.325.426.627.728.930.031.232.333.534.635.837.01011.012.113.214.415.516.617.718.920.021.122.323.424.625.726.928.029.230.331.532.733.835.036.237.438.61112.213.414.515.616.817.919.120.221.422.523.724.826.027.228.329.530.731.933.034.235.436.637.839.040.21213.514.615.816.918.119.320.421.622.823.925.126.327.528.629.831.032.233.434.635.837.038.239.540.741.91314.816.017.118.319.520.621.823.024.225.426.627.829.030.231.432.633.835.036.237.538.739.941.242.443.61416.117.318.519.720.922.123.324.525.726.928.129.330.531.733.034.235.436.737.939.140.441.642.944.245.41517.518.719.921.122.323.524.725.927.228.429.630.932.133.334.635.837.138.439.640.942.243.444.746.047.31618.920.121.322.623.825.026.227.528.730.031.232.533.735.036.337.538.840.141.442.744.045.346.647.949.21720.321.622.824.125.326.627.829.130.331.632.934.135.436.738.039.340.641.943.244.545.947.248.549.851.21821.823.124.325.626.928.129.430.732.033.334.635.937.238.539.841.142.443.845.146.547.849.250.551.953.21923.324.625.927.228.529.831.132.433.735.036.337.639.040.341.643.044.345.747.148.449.851.252.654.055.42024.926.227.528.830.131.532.834.135.436.838.139.540.842.243.644.946.347.749.150.551.953.354.756.157.62126.527.929.230.531.833.234.535.937.338.640.041.442.844.145.546.948.449.851.252.654.155.556.958.459.92228.229.530.932.333.635.036.437.739.140.541.943.344.846.247.649.050.551.953.454.856.357.859.360.862.32329.931.332.734.135.536.838.339.741.142.543.945.446.848.349.751.252.754.255.657.158.660.261.763.264.72431.733.134.535.937.338.840.241.743.144.646.047.549.050.552.053.555.056.558.059.561.162.664.265.867.32533.635.036.437.939.340.842.243.745.246.748.249.751.252.754.355.857.358.960.562.063.665.266.868.470.02635.536.938.439.941.442.844.345.947.448.950.452.053.555.156.758.259.861.463.064.766.367.969.671.272.92737.438.940.442.043.545.046.548.149.651.252.854.456.057.659.260.862.464.165.767.469.170.872.574.275.92839.541.042.644.145.747.348.850.452.053.655.256.958.560.261.863.565.266.968.670.372.073.775.577.379.02941.743.244.846.448.049.651.252.854.556.157.859.561.262.964.666.368.069.871.573.375.176.978.780.582.43043.945.547.148.750.452.053.755.457.158.860.562.264.065.767.569.371.072.974.776.578.380.282.184.085.93146.247.949.551.252.954.656.358.159.861.663.365.166.968.770.572.474.276.178.079.981.883.785.787.689.63248.750.452.153.855.657.359.160.962.764.566.368.270.071.973.875.777.679.581.583.585.487.589.591.593.63351.253.054.856.558.360.262.063.865.767.669.571.473.375.277.279.281.283.285.287.389.391.493.695.797.83453.955.757.659.461.363.165.067.068.970.872.874.876.878.880.882.985.087.189.291.493.595.797.9100.2102.43556.858.660.562.464.466.368.370.372.374.376.378.480.582.684.786.989.191.393.595.798.0100.3102.6105.0107.33659.861.763.765.767.769.771.773.875.978.080.182.384.586.788.991.293.595.898.1100.5102.9105.3107.7110.2112.73762.965.067.069.171.273.375.477.679.882.084.286.588.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.9118.63866.368.470.672.774.977.179.481.683.986.288.691.093.495.898.3100.8103.4105.9108.6111.2113.9116.6119.4122.2125.03970.072.274.476.778.981.383.686.088.490.993.495.998.4101.0103.6106.3109.0111.8114.6117.4120.3123.2126.1129.2132.24073.876.278.580.983.385.788.290.893.395.998.5101.2103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.0140.34178.080.583.085.588.090.693.395.998.7101.4104.3107.1110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.9149.54282.685.287.890.593.296.098.8101.7104.6107.6110.6113.7116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.1160.24387.690.493.296.099.0101.9105.0108.1111.2114.5117.8121.1124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.2172.84493.196.199.1102.2105.4108.6111.9115.3118.7122.3125.9129.6133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.9188.24599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.6143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.2207.546106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.5156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.4233.347114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.4172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.5270.048123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.0193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.0328.249135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6435.2Quanti-TrayおよびDefined Substrate Technologyは、米国および/または他国のIDEXX Laboratories, Inc.の商標または登録商標です。米国特許番号4.925.789; 5.429.933; 5.518.892によって保護されています。他の特許は出願中です。**IDEXX Quanti-Tray/2000 MPN**表大きいウェル陽性数小さいウェル陽性数**25 26** **27 28** **29 30** **31 32** **33 34** **35 36** **37 38** **39 40** **41 42** **43 44** **45 46** **47 48** 025.326.427.428.429.530.531.532.633.634.735.736.837.838.940.041.042.143.144.245.346.347.448.549.5126.627.728.729.830.831.932.934.035.036.137.238.239.340.441.442.543.644.745.746.847.949.050.151.2227.929.030.031.132.233.234.335.436.537.538.639.740.841.943.044.045.146.247.348.449.550.651.752.8329.330.431.432.533.634.735.836.837.939.040.141.242.343.444.545.646.747.848.950.051.252.353.454.5430.731.832.833.935.036.137.238.339.440.541.642.843.945.046.147.248.349.550.651.752.954.055.156.3532.133.234.335.436.537.638.739.941.042.143.244.445.546.647.748.950.051.252.353.554.655.856.958.1633.534.735.836.938.039.240.341.442.643.744.846.047.148.349.450.651.752.954.155.256.457.658.759.9735.036.237.338.439.640.741.943.044.245.346.547.748.850.051.252.353.554.755.957.158.359.460.661.8836.637.738.940.041.242.343.544.745.947.048.249.450.651.853.054.155.356.557.759.060.261.462.663.8938.139.340.541.642.844.045.246.447.648.850.051.252.453.654.856.057.258.459.760.962.163.464.665.81039.740.942.143.344.545.746.948.149.350.651.853.054.255.556.757.959.260.461.762.964.265.466.767.91141.442.643.845.046.347.548.749.951.252.453.754.956.157.458.659.961.262.463.765.066.367.568.870.11243.144.345.646.848.149.350.651.853.154.355.656.858.159.460.762.063.264.565.867.168.469.771.072.41344.946.147.448.649.951.252.553.755.056.357.658.960.261.562.864.165.466.768.069.370.772.073.374.71446.748.049.350.551.853.154.455.757.058.359.660.962.363.664.966.367.668.970.371.673.074.475.777.11548.649.951.252.553.855.156.457.859.160.461.863.164.565.867.268.569.971.372.674.075.476.878.279.61650.551.853.254.555.857.258.559.961.262.664.065.366.768.169.570.972.373.775.176.577.979.380.882.21752.553.955.256.658.059.360.762.163.564.966.367.769.170.571.973.374.876.277.679.180.582.083.584.91854.656.057.458.860.261.663.064.465.867.268.670.171.573.074.475.977.378.880.381.883.384.886.387.81956.858.259.661.062.463.965.366.868.269.771.172.674.175.577.078.580.081.583.184.686.187.689.290.72059.060.461.963.364.866.367.769.270.772.273.775.276.778.279.881.382.884.485.987.589.190.792.293.82161.362.864.365.867.368.870.371.873.374.976.477.979.581.182.684.285.887.489.090.692.293.895.497.12263.865.366.868.369.871.472.974.576.177.679.280.882.484.085.687.288.990.592.193.895.597.198.8100.52366.367.869.471.072.574.175.777.378.980.582.283.885.487.188.790.492.193.895.597.298.9100.6102.4104.12468.970.572.173.775.377.078.680.381.983.685.286.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.92571.773.375.076.678.380.081.783.385.186.888.590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.0111.92674.676.378.079.781.483.184.886.688.490.191.993.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22777.679.481.182.984.686.488.290.091.993.795.597.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.72880.882.684.486.388.189.991.893.795.697.599.4101.3103.3105.2107.2109.2111.2113.2115.2117.3119.3121.4123.5125.62984.286.187.989.891.793.795.697.599.5101.5103.5105.5107.5109.5111.6113.7115.7117.8120.0122.1124.2126.4128.6130.83087.889.791.793.695.697.699.6101.6103.7105.7107.8109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.43191.693.695.697.799.7101.8103.9106.0108.2110.3112.5114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.53295.797.899.9102.0104.2106.3108.5110.7113.0115.2117.5119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133100.0102.2104.4106.6108.9111.2113.5115.8118.2120.5122.9125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434104.7107.0109.3111.7114.0116.4118.9121.3123.8126.3128.8131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.3138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.4145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.3153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.2162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.4173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.1185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.8199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.4216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.8238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.1265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.3302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.3355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.8436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.3574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4866.4920.8980.41046.21119.91203.31299.71413.61553.11732.91986.32419.6>2419.6