(a) Routine monitoring. (1) Public water systems must collect total coliform samples at sites which are representative of water throughout the distribution system according to a written sample siting plan. These plans are subject to State review and revision.
(1) Public water systems must collect total coliform samples at sites which are representative of water throughout the distribution system according to a written sample siting plan. These plans are subject to State review and revision.
(2) The monitoring frequency for total coliforms for community water systems is based on the population served by the system, as follows:
Total Coliform Monitoring Frequency for Community Water Systems------------------------------------------------------------------------
Minimum
number of
Population served samples
per month------------------------------------------------------------------------25 to 1,000 \1\.............................................. 11,001 to 2,500............................................... 22,501 to 3,300............................................... 33,301 to 4,100............................................... 44,101 to 4,900............................................... 54,901 to 5,800............................................... 65,801 to 6,700............................................... 76,701 to 7,600............................................... 87,601 to 8,500............................................... 98,501 to 12,900.............................................. 1012,901 to 17,200............................................. 1517,201 to 21,500............................................. 2021,501 to 25,000............................................. 2525,001 to 33,000............................................. 3033,001 to 41,000............................................. 4041,001 to 50,000............................................. 5050,001 to 59,000............................................. 6059,001 to 70,000............................................. 7070,001 to 83,000............................................. 8083,001 to 96,000............................................. 9096,001 to 130,000............................................ 100130,001 to 220,000........................................... 120220,001 to 320,000........................................... 150320,001 to 450,000........................................... 180450,001 to 600,000........................................... 210600,001 to 780,000........................................... 240780,001 to 970,000........................................... 270970,001 to 1,230,000......................................... 3001,230,001 to 1,520,000....................................... 3301,520,001 to 1,850,000....................................... 3601,850,001 to 2,270,000....................................... 3902,270,001 to 3,020,000....................................... 4203,020,001 to 3,960,000....................................... 4503,960,001 or more............................................ 480------------------------------------------------------------------------\1\ Includes public water systems which have at least 15 service
connections, but serve fewer than 25 persons. If a community water system serving 25 to 1,000 persons has no history of total coliform contamination in its current configuration and a sanitary survey conducted in the past five years shows that the system is supplied solely by a protected groundwater source and is free of sanitary defects, the State may reduce the monitoring frequency specified above, except that in no case may the State reduce the monitoring frequency to less than one sample per quarter. The State must approve the reduced monitoring frequency in writing.
(3) The monitoring frequency for total coliforms for non-community water systems is as follows:
(i) A non-community water system using only ground water (except ground water under the direct influence of surface water, as defined in Sec. 141.2) and serving 1,000 persons or fewer must monitor each calendar quarter that the system provides water to the public, except that the State may reduce this monitoring frequency, in writing, if a sanitary survey shows that the system is free of sanitary defects. Beginning June 29, 1994, the State cannot reduce the monitoring frequency for a non-community water system using only ground water (except ground water under the direct influence of surface water, as defined in Sec. 141.2) and serving 1,000 persons or fewer to less than once/year.
(ii) A non-community water system using only ground water (except ground water under the direct influence of surface water, as defined in Sec. 141.2) and serving more than 1,000 persons during any month must monitor at the same frequency as a like-sized community water system, as specified in paragraph (a)(2) of this section, except the State may reduce this monitoring frequency, in writing, for any month the system serves 1,000 persons or fewer. The State cannot reduce the monitoring frequency to less than once/year. For systems using ground water under the direct influence of surface water, paragraph (a)(3)(iv) of this section applies.
(iii) A non-community water system using surface water, in total or in part, must monitor at the same frequency as a like-sized community water system, as specified in paragraph (a)(2) of this section, regardless of the number of persons it serves.
(iv) A non-community water system using ground water under the direct influence of surface water, as defined in Sec. 141.2, must monitor at the same frequency as a like-sized community water system, as specified in paragraph (a)(2) of this section. The system must begin monitoring at this frequency beginning six months after the State determines that the ground water is under the direct influence of surface water.
(4) The public water system must collect samples at regular time intervals throughout the month, except that a system which uses only ground water (except ground water under the direct influence of surface water, as defined in Sec. 141.2), and serves 4,900 persons or fewer, may collect all required samples on a single day if they are taken from different sites.
(5) A public water system that uses surface water or ground water under the direct influence of surface water, as defined in Sec. 141.2, and does not practice filtration in compliance with Subpart H must collect at least one sample near the first service connection each day the turbidity level of the source water, measured as specified in Sec. 141.74(b)(2), exceeds 1 NTU. This sample must be analyzed for the presence of total coliforms. When one or more turbidity measurements in any day exceed 1 NTU, the system must collect this coliform sample within 24 hours of the first exceedance, unless the State determines that the system, for logistical reasons outside the system's control, cannot have the sample analyzed within 30 hours of collection. Sample results from this coliform monitoring must be included in determining compliance with the MCL for total coliforms in Sec. 141.63.
(6) Special purpose samples, such as those taken to determine whether disinfection practices are sufficient following pipe placement, replacement, or repair, shall not be used to determine compliance with the MCL for total coliforms in Sec. 141.63. Repeat samples taken pursuant to paragraph (b) of this section are not considered special purpose samples, and must be used to determine compliance with the MCL for total coliforms in Sec. 141.63.
(b) Repeat monitoring. (1) If a routine sample is total coliform-positive, the public water system must collect a set of repeat samples within 24 hours of being notified of the positive result. A system which collects more than one routine sample/month must collect no fewer than three repeat samples for each total coliform-positive sample found. A system which collects one routine sample/month or fewer must collect no fewer than four repeat samples for each total coliform-positive sample found. The State may extend the 24-hour limit on a case-by-case basis if the system has a logistical problem in collecting the repeat samples within 24 hours that is beyond its control. In the case of an extension, the State must specify how much time the system has to collect the repeat samples.
(1) If a routine sample is total coliform-positive, the public water system must collect a set of repeat samples within 24 hours of being notified of the positive result. A system which collects more than one routine sample/month must collect no fewer than three repeat samples for each total coliform-positive sample found. A system which collects one routine sample/month or fewer must collect no fewer than four repeat samples for each total coliform-positive sample found. The State may extend the 24-hour limit on a case-by-case basis if the system has a logistical problem in collecting the repeat samples within 24 hours that is beyond its control. In the case of an extension, the State must specify how much time the system has to collect the repeat samples.
(2) The system must collect at least one repeat sample from the sampling tap where the original total coliform-positive sample was taken, and at least one repeat sample at a tap within five service connections upstream and at least one repeat sample at a tap within five service connections downstream of the original sampling site. If a total coliform-positive sample is at the end of the distribution system, or one away from the end of the distribution system, the State may waive the requirement to collect at least one repeat sample upstream or downstream of the original sampling site.
(3) The system must collect all repeat samples on the same day, except that the State may allow a system with a single service connection to collect the required set of repeat samples over a four-day period or to collect a larger volume repeat sample(s) in one or more sample containers of any size, as long as the total volume collected is at least 400 ml (300 ml for systems which collect more than one routine sample/month).
(4) If one or more repeat samples in the set is total coliform-positive, the public water system must collect an additional set of repeat samples in the manner specified in paragraphs (b) (1)-(3) of this section. The additional samples must be collected within 24 hours of being notified of the positive result, unless the State extends the limit as provided in paragraph (b)(1) of this section. The system must repeat this process until either total coliforms are not detected in one complete set of repeat samples or the system determines that the MCL for total coliforms in Sec. 141.63 has been exceeded and notifies the State.
(5) If a system collecting fewer than five routine samples/month has one or more total coliform-positive samples and the State does not invalidate the sample(s) under paragraph (c) of this section, it must collect at least five routine samples during the next month the system provides water to the public, except that the State may waive this requirement if the conditions of paragraph (b)(5) (i) or (ii) of this section are met. The State cannot waive the requirement for a system to collect repeat samples in paragraphs (b) (1)-(4) of this section.
(i) The State may waive the requirement to collect five routine samples the next month the system provides water to the public if the State, or an agent approved by the State, performs a site visit before the end of the next month the system provides water to the public. Although a sanitary survey need not be performed, the site visit must be sufficiently detailed to allow the State to determine whether additional monitoring and/or any corrective action is needed. The State cannot approve an employee of the system to perform this site visit, even if the employee is an agent approved by the State to perform sanitary surveys.
(ii) The State may waive the requirement to collect five routine samples the next month the system provides water to the public if the State has determined why the sample was total coliform-positive and establishes that the system has corrected the problem or will correct the problem before the end of the next month the system serves water to the public. In this case, the State must document this decision to waive the following month's additional monitoring requirement in writing, have it approved and signed by the supervisor of the State official who recommends such a decision, and make this document available to the EPA and public. The written documentation must describe the specific cause of the total coliform-positive sample and what action the system has taken and/or will take to correct this problem. The State cannot waive the requirement to collect five routine samples the next month the system provides water to the public solely on the grounds that all repeat samples are total coliform-negative. Under this paragraph, a system must still take at least one routine sample before the end of the next month it serves water to the public and use it to determine compliance with the MCL for total coliforms in Sec. 141.63, unless the State has determined that the system has corrected the contamination problem before the system took the set of repeat samples required in paragraphs (b) (1)-(4) of this section, and all repeat samples were total coliform-negative.
(6) After a system collects a routine sample and before it learns the results of the analysis of that sample, if it collects another routine sample(s) from within five adjacent service connections of the initial sample, and the initial sample, after analysis, is found to contain total coliforms, then the system may count the subsequent sample(s) as a repeat sample instead of as a routine sample.
(7) Results of all routine and repeat samples not invalidated by the State must be included in determining compliance with the MCL for total coliforms in Sec. 141.63.
(c) Invalidation of total coliform samples. A total coliform-positive sample invalidated under this paragraph (c) does not count towards meeting the minimum monitoring requirements of this section.
(1) The State may invalidate a total coliform-positive sample only if the conditions of paragraph (c)(1) (i), (ii), or (iii) of this section are met.
(i) The laboratory establishes that improper sample analysis caused the total coliform-positive result.
(ii) The State, on the basis of the results of repeat samples collected as required by paragraphs (b) (1) through (4) of this section, determines that the total coliform-positive sample resulted from a domestic or other non-distribution system plumbing problem. The State cannot invalidate a sample on the basis of repeat sample results unless all repeat sample(s) collected at the same tap as the original total coliform-positive sample are also total coliform-positive, and all repeat samples collected within five service connections of the original tap are total coliform-negative (e.g., a State cannot invalidate a total coliform-positive sample on the basis of repeat samples if all the repeat samples are total coliform-negative, or if the public water system has only one service connection).
(iii) The State has substantial grounds to believe that a total coliform-positive result is due to a circumstance or condition which does not reflect water quality in the distribution system. In this case, the system must still collect all repeat samples required under paragraphs (b) (1)-(4) of this section, and use them to determine compliance with the MCL for total coliforms in Sec. 141.63. To invalidate a total coliform-positive sample under this paragraph, the decision with the rationale for the decision must be documented in writing, and approved and signed by the supervisor of the State official who recommended the decision. The State must make this document available to EPA and the public. The written documentation must state the specific cause of the total coliform-positive sample, and what action the system has taken, or will take, to correct this problem. The State may not invalidate a total coliform-positive sample solely on the grounds that all repeat samples are total coliform-negative.
(2) A laboratory must invalidate a total coliform sample (unless total coliforms are detected) if the sample produces a turbid culture in the absence of gas production using an analytical method where gas formation is examined (e.g., the Multiple-Tube Fermentation Technique), produces a turbid culture in the absence of an acid reaction in the Presence-Absence (P-A) Coliform Test, or exhibits confluent growth or produces colonies too numerous to count with an analytical method using a membrane filter (e.g., Membrane Filter Technique). If a laboratory invalidates a sample because of such interference, the system must collect another sample from the same location as the original sample within 24 hours of being notified of the interference problem, and have it analyzed for the presence of total coliforms. The system must continue to re-sample within 24 hours and have the samples analyzed until it obtains a valid result. The State may waive the 24-hour time limit on a case-by-case basis.
(d) Sanitary surveys. (1)(i) Public water systems which do not collect five or more routine samples/month must undergo an initial sanitary survey by June 29, 1994, for community public water systems and June 29, 1999, for non-community water systems. Thereafter, systems must undergo another sanitary survey every five years, except that non-community water systems using only protected and disinfected ground water, as defined by the State, must undergo subsequent sanitary surveys at least every ten years after the initial sanitary survey. The State must review the results of each sanitary survey to determine whether the existing monitoring frequency is adequate and what additional measures, if any, the system needs to undertake to improve drinking water quality.
(1)(i) Public water systems which do not collect five or more routine samples/month must undergo an initial sanitary survey by June 29, 1994, for community public water systems and June 29, 1999, for non-community water systems. Thereafter, systems must undergo another sanitary survey every five years, except that non-community water systems using only protected and disinfected ground water, as defined by the State, must undergo subsequent sanitary surveys at least every ten years after the initial sanitary survey. The State must review the results of each sanitary survey to determine whether the existing monitoring frequency is adequate and what additional measures, if any, the system needs to undertake to improve drinking water quality.
(i) Public water systems which do not collect five or more routine samples/month must undergo an initial sanitary survey by June 29, 1994, for community public water systems and June 29, 1999, for non-community water systems. Thereafter, systems must undergo another sanitary survey every five years, except that non-community water systems using only protected and disinfected ground water, as defined by the State, must undergo subsequent sanitary surveys at least every ten years after the initial sanitary survey. The State must review the results of each sanitary survey to determine whether the existing monitoring frequency is adequate and what additional measures, if any, the system needs to undertake to improve drinking water quality.
(ii) In conducting a sanitary survey of a system using ground water in a State having an EPA-approved wellhead protection program under section 1428 of the Safe Drinking Water Act, information on sources of contamination within the delineated wellhead protection area that was collected in the course of developing and implementing the program should be considered instead of collecting new information, if the information was collected since the last time the system was subject to a sanitary survey.
(2) Sanitary surveys must be performed by the State or an agent approved by the State. The system is responsible for ensuring the survey takes place.
(3) Sanitary surveys conducted by the State under the provisions of Sec. 142.16(o)(2) of this chapter may be used to meet the sanitary survey requirements of this section.
(e) Fecal coliforms/Escherichia coli (E. coli) testing. (1) If any routine or repeat sample is total coliform-positive, the system must analyze that total coliform-positive culture medium to determine if fecal coliforms are present, except that the system may test for E. coli in lieu of fecal coliforms. If fecal coliforms or E. coli are present, the system must notify the State by the end of the day when the system is notified of the test result, unless the system is notified of the result after the State office is closed, in which case the system must notify the State before the end of the next business day.
(1) If any routine or repeat sample is total coliform-positive, the system must analyze that total coliform-positive culture medium to determine if fecal coliforms are present, except that the system may test for E. coli in lieu of fecal coliforms. If fecal coliforms or E. coli are present, the system must notify the State by the end of the day when the system is notified of the test result, unless the system is notified of the result after the State office is closed, in which case the system must notify the State before the end of the next business day.
(2) The State has the discretion to allow a public water system, on a case-by-case basis, to forgo fecal coliform or E. coli testing on a total coliform-positive sample if that system assumes that the total coliform-positive sample is fecal coliform-positive or E. coli-positive. Accordingly, the system must notify the State as specified in paragraph (e)(1) of this section and the provisions of Sec. 141.63(b) apply.
(f) Analytical methodology. (1) The standard sample volume required for total coliform analysis, regardless of analytical method used, is 100 ml.
(1) The standard sample volume required for total coliform analysis, regardless of analytical method used, is 100 ml.
(2) Public water systems need only determine the presence or absence of total coliforms; a determination of total coliform density is not required.
(3) Public water systems must conduct total coliform analyses in accordance with one of the analytical methods in the following table or one of the alternative methods listed in appendix A to subpart C of this part. ----------------------------------------------------------------------------------------------------------------
Organism Methodology \12\ Citation \1\----------------------------------------------------------------------------------------------------------------Total Coliforms \2\................... Total Coliform Fermentation Technique \3 9221A, B.
4 5\.
Total Coliform Membrane Filter Technique 9222A, B, C.
\6\.
Presence-Absence (P-A) Coliform Test \5 9221D.
7\.
ONPG-MUG Test \8\....................... 9223.
Colisure Test. \9\
E*Colite [supreg] Test. \10\
m-ColiBlue24 [supreg] Test. \11\
Readycult [supreg] Coliforms 100
Presence/Absence Test. \13\
Membrane Filter Technique using
Chromocult [supreg] Coliform Agar. \14\
Colitag [supreg] Test. \15\----------------------------------------------------------------------------------------------------------------The procedures shall be done in accordance with the documents listed below. The incorporation by reference of
the following documents listed in footnotes 1, 6, 8, 9, 10 , 11, 13, 14 and 15 was approved by the Director of
the Federal Register in accordance with 5 U.S.C. 552(a) and 1 CFR part 51. Copies of the documents may be
obtained from the sources listed below. Information regarding obtaining these documents can be obtained from
the Safe Drinking Water Hotline at 800-426-4791. Documents may be inspected at EPA's Drinking Water Docket,
EPA West, 1301 Constitution Avenue, NW., EPA West, Room B102, Washington DC 20460 (Telephone: 202-566-2426);
or at the National Archives and Records Administration (NARA). For information on the availability of this
material at NARA, call 202-741-6030, or go to: http://www.archives.gov/federal--register/code--of--federal--
regulations/ibr--locations.html.
\1\ Standard Methods for the Examination of Water and Wastewater, 18th edition (1992), 19th edition (1995), or
20th edition (1998). American Public Health Association, 1015 Fifteenth Street, NW., Washington, DC 20005. The
cited methods published in any of these three editions may be used. In addition, the following online versions
may also be used: 9221 A, B, D-99, 9222 A, B, C-97, and 9223 B-97. Standard Methods Online are available at
http://www.standardmethods.org. The year in which each method was approved by the Standard Methods Committee
is designated by the last two digits in the method number. The methods listed are the only Online versions
that may be used.\2\ The time from sample collection to initiation of analysis may not exceed 30 hours. Systems are encouraged
but not required to hold samples below 10 deg. C during transit.\3\ Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system
conducts at least 25 parallel tests between this medium and lauryl tryptose broth using the water normally
tested, and this comparison demonstrates that the false-positive rate and false-negative rate for total
coliform, using lactose broth, is less than 10 percent.\4\ If inverted tubes are used to detect gas production, the media should cover these tubes at least one-half to
two-thirds after the sample is added.\5\ No requirement exists to run the completed phase on 10 percent of all total coliform-positive confirmed
tubes.\6\ MI agar also may be used. Preparation and use of MI agar is set forth in the article, ``New medium for the
simultaneous detection of total coliform and Escherichia coli in water'' by Brenner, K.P., et. al., 1993,
Appl. Environ. Microbiol. 59:3534-3544. Also available from the Office of Water Resource Center (RC-4100T),
1200 Pennsylvania Avenue, NW., Washington, DC 20460, EPA/600/J-99/225. Verification of colonies is not
required.\7\ Six-times formulation strength may be used if the medium is filter-sterilized rather than autoclaved.\8\ The ONPG-MUG Test is also known as the Autoanalysis Collect System.\9\ A description of the Colisure Test, Feb 28, 1994, may be obtained from IDEXX Laboratories, Inc., One IDEXX
Drive, Westbrook, Maine 04092. The Colisure Test may be read after an incubation time of 24 hours.\10\ A description of the E*Colite [supreg] Test, ``Presence/Absence for Coliforms and E. Coli in Water,'' Dec
21, 1997, is available from Charm Sciences, Inc., 36 Franklin Street, Malden, MA 02148-4120.\11\ A description of the m-ColiBlue24 [supreg] Test, Aug 17, 1999, is available from the Hach Company, 100
Dayton Avenue, Ames, IA 50010.\12\ EPA strongly recommends that laboratories evaluate the false-positive and negative rates for the method(s)
they use for monitoring total coliforms. EPA also encourages laboratories to establish false-positive and
false-negative rates within their own laboratory and sample matrix (drinking water or source water) with the
intent that if the method they choose has an unacceptable false-positive or negative rate, another method can
be used. The Agency suggests that laboratories perform these studies on a minimum of 5% of all total coliform-
positive samples, except for those methods where verification/confirmation is already required, e.g., the M-
Endo and LES Endo Membrane Filter Tests, Standard Total Coliform Fermentation Technique, and Presence-Absence
Coliform Test. Methods for establishing false-positive and negative-rates may be based on lactose
fermentation, the rapid test for [beta]-galactosidase and cytochrome oxidase, multi-test identification
systems, or equivalent confirmation tests. False-positive and false-negative information is often available in
published studies and/or from the manufacturer(s).\13\ The Readycult [supreg] Coliforms 100 Presence/Absence Test is described in the document, ``Readycult
[supreg] Coliforms 100 Presence/Absence Test for Detection and Identification of Coliform Bacteria and
Escherichla coli in Finished Waters'', November 2000, Version 1.0, available from EM Science (an affiliate of
Merck KGgA, Darmstadt Germany), 480 S. Democrat Road, Gibbstown, NJ 08027-1297. Telephone number is (800) 222-
0342, e-mail address is: adellenbusch@emscience.com.\14\ Membrane Filter Technique using Chromocult [supreg] Coliform Agar is described in the document,
``Chromocult [supreg] Coliform Agar Presence/Absence Membrane Filter Test Method for Detection and
Identification of Coliform Bacteria and Escherichla coli in Finished Waters'', November 2000, Version 1.0,
available from EM Science (an affiliate of Merck KGgA, Darmstadt Germany), 480 S. Democrat Road, Gibbstown, NJ
08027-1297. Telephone number is (800) 222-0342, e-mail address is: adellenbusch@emscience.com.\15\ Colitag [supreg] product for the determination of the presence/absence of total coliforms and E. coli is
described in ``Colitag [supreg] Product as a Test for Detection and Identification of Coliforms and E. coli
Bacteria in Drinking Water and Source Water as Required in National Primary Drinking Water Regulations,''
August 2001, available from CPI International, Inc., 5580 Skylane Blvd., Santa Rosa, CA, 95403, telephone
(800) 878-7654, Fax (707) 545-7901, Internet address http://www.cpiinternational.com.
(4) [Reserved]
(5) Public water systems must conduct fecal coliform analysis in accordance with the following procedure. When the MTF Technique or Presence-Absence (PA) Coliform Test is used to test for total coliforms, shake the lactose-positive presumptive tube or P-A vigorously and transfer the growth with a sterile 3-mm loop or sterile applicator stick into brilliant green lactose bile broth and EC medium to determine the presence of total and fecal coliforms, respectively. For EPA-approved analytical methods which use a membrane filter, transfer the total coliform-positive culture by one of the following methods: remove the membrane containing the total coliform colonies from the substrate with a sterile forceps and carefully curl and insert the membrane into a tube of EC medium (the laboratory may first remove a small portion of selected colonies for verification), swab the entire membrane filter surface with a sterile cotton swab and transfer the inoculum to EC medium (do not leave the cotton swab in the EC medium), or inoculate individual total coliform-positive colonies into EC Medium. Gently shake the inoculated tubes of EC medium to insure adequate mixing and incubate in a waterbath at 44.5 0.2 [deg]C for 24 2 hours. Gas production of any amount in the inner fermentation tube of the EC medium indicates a positive fecal coliform test. The preparation of EC medium is described in Method 9221E (paragraph 1a) in Standard Methods for the Examination of Water and Wastewater, 18th edition (1992), 19th edition (1995), and 20th edition (1998); the cited method in any one of these three editions may be used. Public water systems need only determine the presence or absence of fecal coliforms; a determination of fecal coliform density is not required.
(6) Public water systems must conduct analysis of Escherichia coli in accordance with one of the following analytical methods or one of the alternative methods listed in appendix A to subpart C of this part.
(i) EC medium supplemented with 50 [micro]g/mL of 4-methylumbelliferyl-beta-D-glucuronide (MUG) (final concentration), as described in Method 9222G in Standard Methods for the Examination of Water and Wastewater, 19th edition (1995) and 20th edition (1998). Either edition may be used. Alternatively, the 18th edition (1992) may be used if at least 10 mL of EC medium, as described in paragraph (f)(5) of this section, is supplemented with 50 [micro]g/mL of MUG before autoclaving. The inner inverted fermentation tube may be omitted. If the 18th edition is used, apply the procedure in paragraph (f)(5) of this section for transferring a total coliform-positive culture to EC medium supplemented with MUG, incubate the tube at 44.5 0.2 [deg]C for 24 2 hours, and then observe fluorescence with an ultraviolet light (366 nm) in the dark. If fluorescence is visible, E. coli are present.
(ii) Nutrient agar supplemented with 100 [micro]g/mL of 4-methylumbelliferyl-beta-D-glucuronide (MUG) (final concentration), as described in Method 9222G in Standard Methods for the Examination of Water and Wastewater, 19th edition (1995) and 20th edition (1998). Either edition may be used for determining if a total coliform-positive sample, as determined by a membrane filter technique, contains E. coli. Alternatively, the 18th edition (1992) may be used if the membrane filter containing a total coliform-positive colony(ies) is transferred to nutrient agar, as described in Method 9221B (paragraph 3) of Standard Methods (18th edition), supplemented with 100 [micro]g/mL of MUG. If the 18th edition is used, incubate the agar plate at 35 [deg]C for 4 hours and then observe the colony(ies) under ultraviolet light (366 nm) in the dark for fluorescence. If fluorescence is visible, E. coli are present.
(iii) Minimal Medium ONPG-MUG (MMO-MUG) Test, as set forth in the article ``National Field Evaluation of a Defined Substrate Method for the Simultaneous Detection of Total Coliforms and Escherichia coli from Drinking Water: Comparison with Presence-Absence Techniques'' (Edberg et al.), Applied and Environmental Microbiology, Volume 55, pp. 1003-1008, April 1989. (Note: The Autoanalysis Colilert System is an MMO-MUG test). If the MMO-MUG test is total coliform-positive after a 24-hour incubation, test the medium for fluorescence with a 366-nm ultraviolet light (preferably with a 6-watt lamp) in the dark. If fluorescence is observed, the sample is E. coli-positive. If fluorescence is questionable (cannot be definitively read) after 24 hours incubation, incubate the culture for an additional four hours (but not to exceed 28 hours total), and again test the medium for fluorescence. The MMO-MUG Test with hepes buffer in lieu of phosphate buffer is the only approved formulation for the detection of E. coli.
(iv) The Colisure Test. A description of the Colisure Test may be obtained from the Millipore Corporation, Technical Services Department, 80 Ashby Road, Bedford, MA 01730.
(v) The membrane filter method with MI agar, a description of which is cited in footnote 6 to the table in paragraph (f)(3) of this section.
(vi) E*Colite [supreg] Test, a description of which is cited in footnote 10 to the table at paragraph (f)(3) of this section.
(vii) m-ColiBlue24 [supreg] Test, a description of which is cited in footnote 11 to the table in paragraph (f)(3) of this section.
(viii) Readycult [supreg] Coliforms 100 Presence/Absence Test, a description of which is cited in footnote 13 to the table at paragraph (f)(3) of this section.
(ix) Membrane Filter Technique using Chromocult [supreg] Coliform Agar, a description of which is cited in footnote 14 to the table at paragraph (f)(3) of this section.
(x) Colitag [supreg], a description of which is cited in footnote 15 to the table at paragraph (f)(3) of this section.
(7) As an option to paragraph (f)(6)(iii) of this section, a system with a total coliform-positive, MUG-negative, MMO-MUG test may further analyze the culture for the presence of E. coli by transferring a 0.1 ml, 28-hour MMO-MUG culture to EC Medium + MUG with a pipet. The formulation and incubation conditions of EC Medium + MUG, and observation of the results are described in paragraph (f)(6)(i) of this section.
(8) The following materials are incorporated by reference in this section with the approval of the Director of the Federal Register in accordance with 5 U.S.C. 552(a) and 1 CFR part 51. Copies of the analytical methods cited in Standard Methods for the Examination of Water and Wastewater (18th, 19th, and 20th editions) may be obtained from the American Public Health Association et al.; 1015 Fifteenth Street, NW., Washington, DC 20005-2605. Copies of the MMO-MUG Test, as set forth in the article ``National Field Evaluation of a Defined Substrate Method for the Simultaneous Enumeration of Total Coliforms and Escherichia coli from Drinking Water: Comparison with the Standard Multiple Tube Fermentation Method'' (Edberg et al.) may be obtained from the American Water Works Association Research Foundation, 6666 West Quincy Avenue, Denver, CO 80235. Copies of the MMO-MUG Test as set forth in the article ``National Field Evaluation of a Defined Substrate Method for the Simultaneous Enumeration of Total Coliforms and Escherichia coli from Drinking Water: Comparison with the Standard Multiple Tube Fermentation Method'' (Edberg et al.) may be obtained from the American Water Works Association Research Foundation, 6666 West Quincy Avenue, Denver, CO 80235. A description of the Colisure Test may be obtained from the Millipore Corp., Technical Services Department, 80 Ashby Road, Bedford, MA 01730. Copies may be inspected at EPA's Drinking Water Docket; 401 M St., SW.; Washington, DC 20460, or at the National Archives and Records Administration (NARA). For information on the availability of this material at NARA, call 202-741-6030, or go to: http://www.archives.gov/federal--register/code--of--federal--regulations/ibr--locations.html.
(g) Response to violation. (1) A public water system which has exceeded the MCL for total coliforms in Sec. 141.63 must report the violation to the State no later than the end of the next business day after it learns of the violation, and notify the public in accordance with subpart Q.
(1) A public water system which has exceeded the MCL for total coliforms in Sec. 141.63 must report the violation to the State no later than the end of the next business day after it learns of the violation, and notify the public in accordance with subpart Q.
(2) A public water system which has failed to comply with a coliform monitoring requirement, including the sanitary survey requirement, must report the monitoring violation to the State within ten days after the system discovers the violation, and notify the public in accordance with subpart Q.
(h) The provisions of paragraphs (a) and (d) of this section are applicable until March 31, 2016. The provisions of paragraphs (b), (c), (e), (f), and (g) of this section are applicable until all required repeat monitoring under paragraph (b) of this section and fecal coliform or E. coli testing under paragraph (e) of this section that was initiated by a total coliform-positive sample taken before April 1, 2016 is completed, as well as analytical method, reporting, recordkeeping, public notification, and consumer confidence report requirements associated with that monitoring and testing. Beginning April 1, 2016, the provisions of subpart Y of this part are applicable, with systems required to begin regular monitoring at the same frequency as the system-specific frequency required on March 31, 2016. [54 FR 27562, June 29, 1989]
Editorial Note: For Federal Register citations affecting Sec. 141.21, see the List of CFR Sections Affected, which appears in the Finding Aids section of the printed volume and at www.fdsys.gov.