Water Bacteriology Laboratory Coliforms and E. Coli 24 Hours or Less

1. Arkansas Department of Health Water Bacteriology Laboratory Coliforms and E. Coli 24 Hours or Less Water Bacteriology Laboratory Coliforms and E. Coli 24 Hours or Less

2. ANCIENT HAMLET SECRETUPON TRAVELING THRU MILWAUKEE

3. Colilert - Defined SubstrateTechnology • Colilert uses the patented Defined Substrate Technology (DST) to simultaneously detect and confirm total coliforms and E. Coli. Two nutrient-indicators, ONPG and MUG, are the major sources of carbon in Colilert and can be metabolized by the Coliform enzyme B-galactosidase and the E. Coli enzyme B-glucuronidase, respectively.

4. As coliforms grow in colilert, they use B-galactosidase to metabolize ONPG and change it from clear to yellow. E. coli uses B-glucuronidase to metabolize MUG and create fluorescence. Since most non-coliforms that do have these enzymes are selectively suppressed by Colilert’s specifically formulated matrix.

5. This approach is different from traditional media, which provide a nutrient-rich environment that supports the growth of both target organisms and non-target. When non-targets grow and mimic target organisms, false positives occur. Growth of non-targets can also suppress target organisms and give false negatives in traditional media.

6. To suppress non-targets, traditional media often include high levels of salts, detergents, or other selective agents - which may inadvertently suppress target organisms and give further false negatives.

7. Two Ways to Use ColilertPresence/Absence (P/A) Proven: • The only US EPA-approved, 24-hour test for drinking and source waters • International approvals include US Standard Methods, AOAC, IBWA, and NCIMS. • Millions of tests are run worldwide every year Add reagent to sample and incubate 24 hours.

8. Read Results: • Clear = Negative • Yellow = total Coliforms • Yellow/fluorescent = E. coli Yellow = Positive Total Coliforms Blue= Positive Escherichiacoli

9. QUANTIFICATION EASY • Ease of use simplifies training • Unit-dosed packaging eliminates media preparation • No repeat testing due to clogged filters or heterotrophic interference • QC procedure can be done in 15 minutes per month Add reagent to sample

10. RAPID • Under one minute hands-on time • Detects total coliforms and E.coli simultaneously in 24 hours or less • No confirmations needed • No glassware cleaning or colony counting • Note: Colilert - 18 takes just 18 hours Pour into Quanti-Tray (counts from1 to 200) or Quanti-Tray/2000 (counts from 1 to 2,419).

11. Yellow = Positive! Seal in Quanti-Tray Sealer and Place in incubator for 24 hours

12. Read Quanti-Tray results: • Yellow wells = total coliforms • Yellow/fluorescentwells = E.coli OR

13. CON’T • Read Quanti-Tray/2000 results: • Yellowwells = total coliforms • Yellow/fluorescent wells = E.coli

20. Logging In Samples

23. Colilert Reagent

24. Step 1

25. Step 2

26. Step 3

27. Flexible • A Colilert Snap Pak can be used for P/A or quantification testing • Quanti-Tray provides counts to 200 cfu/100 ml without dilutions • Quanti-Tray/2000 provides counts to 2,419 cfu/100 ml without dilutions • Colilert is also available predispensed in 10 ml MPN tubes

28. Quanti - Tray Sealer Used for raw water analysis

29. Samples Are Incubated for 24 Hours

30. Samples During Incubation Yellow discoloration indicates presence of Total Coliforms

34. Vessels Broken In Transit 02/02/92 to 04/03/98 Out of 10,693 samples received , 150 leaked due to broken bottles. Most of these occurred the first week.

35. For the same period last year, 29 of the old bottles leaked from all causes out of 10,882 samples received.

36. Samples with color may interfere with color change rendering the result indeterminate. 6 samples were indeterminate due to inherent color.

37. When sample contains excess chlorine - Stage 1

38. Stage 2 Foaming occurs upon addition of reagent.

39. Stage 341 indeterminate samples due to excess chlorine Blue Ring at the top of the sample vessel indicates excess chlorine

40. 02/03/97 - 04/03/97 10,882 samples rec’d MF 133 TCP 33 FCP 230 TNTCs 41 CGs 02/02/98 - 04/03/98 10,693 samples rec’d colilert 247 TCP 53 ECP MF 28 TCP 11 FCP Positive Results

41. Accurate . • Identifies E.Coli specifically, which eliminates unneeded public notification due to Klebsiella pneumoniae • Suppresses up to 2 million heterotrophs per 100 mL • Eliminates the subjective interpretation found in traditional methods • Detects a single viable coliform or E.coli per sample

42. Economical • Minimizes evening and weekend work • Up to 12-month shelf life at room temperature • 20 to 50% lessexpensive than traditional methods • 95% lessequipment costs

43. Water Bacteriology Laboratory Rusty Baldwin Micro II James Owens Micro II Cathy Moore Micro II Eloise Glover Lab Tech II Eve Berry Micro II DeLois Manor Supervisor

45. 1998 Cathy Walker TechnicalStaff, Lab Administration