Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results –...

40
Coliform Sample Collection Module 4 Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Transcript of Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results –...

Page 1: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Coliform Sample Collection

Module 4

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 2: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Learning Objectives

• Be able to take and explain how to take a good coliform sample

• Recognize the challenges in taking a good sample

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 3: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Coliform Sampling – Why? • Indicator of pathogen

contamination • Total Coliform (TC)

– Not necessarily a health threat in itself; used to indicate other potentially harmful bacteria

– A very common microbe – Should be absent if chlorine residual is

adequate. • E. coli

– A subset of total coliform which indicates fecal waste contamination from mammals (humans, cows, etc)

– Found only in mammal feces

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 4: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Coliform Sampling – Best Practices • Collecting total coliform samples correctly

and properly is absolutely critical in protecting public health

• Improper sampling is the most common reason for positive results (false positive) – Repeated sampling requires extra effort, time,

and money – May lead to unnecessary MCL violation and

subsequent corrective measures

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 5: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Sampling Procedures 1. Assemble sampling supplies 2. Go to sampling location(s) specified in the

sampling plan 3. Remove any aerators, strainers, or hoses from

the tap 4. Open the cold water tap for about 2 to 3 minutes

before collecting the sample 5. Fill out label, tag, and lab form in waterproof ink 6. Adjust the flow to about the width of a pencil

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 6: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Sampling Procedures, cont.

7. Remove the bottle cap 8. Fill the bottle to the shoulder or about ¼ inch from the top 9. Place the cap on the bottle and screw it down tightly 10. Turn the tap off and replace the aerator, strainer, or hose 11. Check the information on the label 12. Complete any additional lab forms that come with the

sample bottle 13. Refrigerate or ice the samples; samples much reach the

lab for processing within 30 hours of sample collection

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 7: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

1. Assemble Supplies

• 125 ml sterilized plastic bottles and bottle rack • Dechlorination agent (do not rinse out bottle) • Cooler with ice or dry ice • Label and lab form (chain of custody form) • Pen / Sharpee

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 8: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Wash Your Hands! THINK STERILE! Assume your hands are dirty even after you wash them…

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 9: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Preparation and Handling

• Wear clean clothing, wash your hands and try to keep them bacteria free for the collection process

• Watch for contamination sources nearby activities – soil disturbances- sewer lift stations –

animals/manure

• Avoid talking and disturbing the air while collecting (sneezing/coughing)

• Smoking during sample collection is not advised. If it is TC+ it will be you who has to recollect

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 10: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

2. Go to Location(s) in Sampling Plan - Sample Tap - Do’s

• Tap should be clean, in good shape, and free of attachments

• Sample cold water only – Valves that control hot and cold independently – Water heaters can be laden with bacteria

• Use a line directly connected to the main • Sample indoors, when possible

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 11: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Sampling Taps –DON’Ts • Sample tap should NOT be:

– Outdoors – Too close to the bottom of the sink – Swivel-type with a single valve for both hot and cold

water – Leaking or on a leaky pipe – Threaded in the interior – Upward flowing (e.g. drinking fountains) – Located in a room of questionable sanitary conditions – Attached to any household point-of-entry or point-of-use

devices (e.g. aerators) – Drinking fountains

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 12: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

What to Avoid • Faucets to avoid:

– Swivel-type faucets that have a single valve for hot and cold water

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 13: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

What to Avoid

– Outdoor faucets – Faucets close to or below ground level

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 14: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

What to Avoid – Faucets that point upward

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 15: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

What to Avoid – Faucets in places highly prone to

contaminations (e.g. janitor’s closet, public rest rooms)

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 16: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

3. Remove Aerator, Strainer, or Hose

• Can trap sediment or particulates

• Biofilms can form in a hose

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 17: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

4. Open Cold Water for 2-3 Minutes

• Want to get water representative of conditions in the water main

• When temperature stabilizes is a good guide

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 18: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

5. Fill out Label, Tag, and Lab Form

• In waterproof ink

• Write clearly

• Write the Date and Time sample collected

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 19: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

6. Adjust Flow to Width of a Pencil

• You want a steady, controlled flow

• Don’t change the flow once you start sampling (could dislodge microbial growth)

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 20: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

7. Remove the Bottle Cap • Be careful not to touch the inside of the

bottle or bottle cap.

• Do not lay the cap down or put it in your pocket.

• STERILE, STERILE, STERILE!!!!

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 21: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

8. Fill Bottle to the Shoulder, ¼ Inch From the Top

Don’t rinse the bottle

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 22: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

9. Place Cap on Bottle and Screw it Down Tightly

Think, STERILE

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 23: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

10. Turn the Tap Off and Replace the Aerator, Strainer,

or Hose

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 24: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

11. Check the Information on the Label

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 25: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

12. Complete any Additional Lab Forms

• Chain of custody • Make sure to write clearly in ink • Who Transported Sample • Date and Time of Delivery/drop off • Number of Samples dropped off

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 26: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Developed by American Water Works Association with funds from the U.S. Environmental Protection Agency, Published 2015

Box #1 Matrix Types Box #2 Preservative

City of Chandler DW = Drinking Water A = HClWater Treatment Plant Laboratory WW = Wastew ater B = HNO3

1475 E. Pecos Road SW = Surface Water C = H2SO4

Chandler, Arizona 85225 GW = Groundw ater D = NaOHPhone (480) 782-3655 or (480) 782-3659 or (480) 782-3656 S = Soil/Solid E = Ice Only

T = Travel Blank F = Other________________

Project Name: TCR G = Sludge

Project Manager: Anupa Jain O = Other_______________

Sampled By: Matt Dregely

Date/Time Sampled Remarks

Location 1 X DW 2 X

Location 2 X DW 1 X

Comments/Special Instructions:

Date/TimePrint Print

Signature Signature

Print Print

Signature Signature

Total No. of Containers: Temperature: Samples received on ice? Yes No

Received by

1

2

Relinquished by Date/Time

LaboratoryIdentification

NumberCom

posit

e

Gra

b

Mat

rix (B

ox #

1)

Pres

erva

tive

(Box

#2)

# of

Con

taine

rs

CHAIN OF CUSTODY RECORD

Client'sSample Identification and/or

Sample Location

REQUESTED ANALYSES

Tota

l Col

iform

s and

E.C

oli

Page 27: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

13. Ice and Send to Lab for Processing Within 30 Hours

• Refrigeration recommended; Cooler with blue ice

• The quicker it gets to the lab the better • Use a certified laboratory for analysis

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 28: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal
Page 29: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal
Page 30: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal
Page 31: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Helpful Hints

• Sample early in the week or month

• If you feel something went wrong, resample – Bottles are cheap, but false positive samples

are not

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 32: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Common Issues that can lead to undesired results

Improper Sampling Techniques

• Not Flushing the Tap

• Improper Handling of Bags

• Exceed 30 Hour Holding Time

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 33: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Avoid Sampling in the Rain

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 34: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Keep your faucets maintained – no spray

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 35: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Who’s Responsible??

• The WATER SYSTEM PERSONNEL are responsible for insuring that all water samples are collected during the correct compliance period

• Utility responsible that the results go to the regulatory agency

• Violation occurs if no sample taken or reported – Includes Public Notice and other measures

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 36: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Laboratory Results • You will be notified by Region/District or

Lab if you have a TC+ Sample • Collect Repeats and Triggered Source

samples within 24 hours or as scheduled • May require corrective action be taken to

resolve contamination

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 37: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Colilert Test 1. Collect proper sample 2. Add one sample pack 3. Cap and shake 4. Incubate at 35oC for 24 hours 5. Read results

– Negative - Less yellow than comparator – Positive total coliform – Yellow equal or

greater – Positive E. coli – yellow and fluorescence

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 38: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

Online Resources

• A Small Systems Guide to the Total Coliform Rule

http://www.epa.gov/ogwdw/disinfection/tcr/pdfs/small-tcr.pdf

• AWWA Video: Reliable Coliform Sampling for Water Systems http://www.awwa.org/store/productdetail.aspx?productid=7089

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

http://www.epa.gov/ogwdw/disinfection/tcr/pdfs/small-tcr.pdf
http://www.epa.gov/ogwdw/disinfection/tcr/pdfs/small-tcr.pdf
http://www.awwa.org/store/productdetail.aspx?productid=7089
http://www.awwa.org/store/productdetail.aspx?productid=7089
Page 39: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

QUESTIONS???

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015

Page 40: Coliform Sample Collection3. Cap and shake 4. Incubate at 35 o C for 24 hours 5. Read results – Negative - Less yellow than comparator – Positive total coliform – Yellow equal

LUNCH

Developed by AWWA in partnership with RCAP and funded by USEPA, Published 2015


Sample Siting Plans and Coliform Sample Collection...Coliform Sample Collection • If the coliform sample is rejected by your lab: A replacement sample is required within 24 hours

Sample Siting Plans and Coliform Sample Collection...Coliform Sample Collection • If the coliform sample is rejected by your lab: A replacement sample is required within 24 hours

Method Comparison: Analysis of Fecal Coliform Samples … · Analysis of Fecal Coliform Samples . from Washington State ... Colilert®-18 (CS ... Analysis of Fecal Coliform Samples

Method Comparison: Analysis of Fecal Coliform Samples … · Analysis of Fecal Coliform Samples . from Washington State ... Colilert®-18 (CS ... Analysis of Fecal Coliform Samples

Issaquah Creek Basin Water Cleanup Plan for Fecal Coliform ... · forming units per 100 milliliters of water (cfu/100mL). Fecal Coliform (FC): Fecal coliform is bacteria present in

Issaquah Creek Basin Water Cleanup Plan for Fecal Coliform ... · forming units per 100 milliliters of water (cfu/100mL). Fecal Coliform (FC): Fecal coliform is bacteria present in

Food Microbiology - Livsmedelsverket ·  · 2015-11-26• Coliform bacteria 30 ºC ... • Thermotolerant coliform bacteria • Escherichia coli • Presumptive Bacillus cereus •

Food Microbiology - Livsmedelsverket ·  · 2015-11-26• Coliform bacteria 30 ºC ... • Thermotolerant coliform bacteria • Escherichia coli • Presumptive Bacillus cereus •

Coliform Bacteria

Coliform Bacteria

Implementation Guidance for the Fecal Coliform Total Daily ...Appendix A, Fecal Coliform Criteria and Information, discusses the fecal coliform criteria and provides background information

Implementation Guidance for the Fecal Coliform Total Daily ...Appendix A, Fecal Coliform Criteria and Information, discusses the fecal coliform criteria and provides background information

The Revised Total Coliform Rule - US EPA...repeat sample or . E. coli-positive repeat sample, or any total coliform-positive repeat sample following a fecal coliform-positive or .

The Revised Total Coliform Rule - US EPA...repeat sample or . E. coli-positive repeat sample, or any total coliform-positive repeat sample following a fecal coliform-positive or .

TM Rev.1 / 22.06.2016 I F U ChromaticTM Coli Coliform · ChromaticTM Coli Coliform Chromogenic medium for detection and enumeration of Escherichia coli and coliform bacteria in food

TM Rev.1 / 22.06.2016 I F U ChromaticTM Coli Coliform · ChromaticTM Coli Coliform Chromogenic medium for detection and enumeration of Escherichia coli and coliform bacteria in food

South Branch Yellow Medicine River Fecal Coliform Total ... · 1 Yellow Medicine River Feed Lot Survey, 2000, Appendix 5 2 Robert Olson, Lincoln County Environmental Services. 3 Chris

South Branch Yellow Medicine River Fecal Coliform Total ... · 1 Yellow Medicine River Feed Lot Survey, 2000, Appendix 5 2 Robert Olson, Lincoln County Environmental Services. 3 Chris

Coliform standards in India

Coliform standards in India

APPENDIX A FECAL COLIFORM AND E. COLI SAMPLING ATA …...Draft Final Project Report Appendix A TMDLs for Fecal coliform in Corralitos and Salsipuedes Creeks 1 APPENDIX A FECAL COLIFORM

APPENDIX A FECAL COLIFORM AND E. COLI SAMPLING ATA …...Draft Final Project Report Appendix A TMDLs for Fecal coliform in Corralitos and Salsipuedes Creeks 1 APPENDIX A FECAL COLIFORM

Coliform Es

Coliform Es

Ponca Creek Fecal Coliform TMDL - DENR · based on the chronic standard for fecal coliform. The fecal coliform criteria for the limited contact recreation beneficial use requires

Ponca Creek Fecal Coliform TMDL - DENR · based on the chronic standard for fecal coliform. The fecal coliform criteria for the limited contact recreation beneficial use requires

Wentachee River Watershed Fecal Coliform Bacteria TMDL · Fecal coliform bacteria data show that all three creeks had higher fecal coliform bacteria concentrations and loads during

Wentachee River Watershed Fecal Coliform Bacteria TMDL · Fecal coliform bacteria data show that all three creeks had higher fecal coliform bacteria concentrations and loads during

Saturday Night Club Music Collection3 - · PDF fileSaturday Night Club Music Collection3 ... Bach,Johann Sebastian Gavotte and Musette fr. 3rd English Suite Oliver ... Drigo,Richard

Saturday Night Club Music Collection3 - · PDF fileSaturday Night Club Music Collection3 ... Bach,Johann Sebastian Gavotte and Musette fr. 3rd English Suite Oliver ... Drigo,Richard

Blue Earth River Basin Fecal Coliform TMDL Report · e. Estimated Fecal Coliform Bacteria Produced by Humans and Animals f. Estimated Fecal Coliform Bacteria Produced by Source Category

Blue Earth River Basin Fecal Coliform TMDL Report · e. Estimated Fecal Coliform Bacteria Produced by Humans and Animals f. Estimated Fecal Coliform Bacteria Produced by Source Category

Revised Total Coliform Rule Presentation · Revised Total Coliform Rule WARWS Conference September 2015 * Actual bumper sticker seen at the last conference. Revised Total Coliform

Revised Total Coliform Rule Presentation · Revised Total Coliform Rule WARWS Conference September 2015 * Actual bumper sticker seen at the last conference. Revised Total Coliform

TMDL – Fecal coliform

TMDL – Fecal coliform

Total Coliform

Total Coliform

Revised Total Coliform Rule

Revised Total Coliform Rule