Water Microbiology Laboratory Evaluation Form - Illinois Department Of Public Health Page 18
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22WATER MICROBIOLOGY LABORATORY EVALUATION FORM
State of Illinois
Illinois Department of Public Health
METHODOLOGY (cont’d)
j.
Total Coliform-positive samples exposed to UV light (365-366nm); blue fluorescence, E. coli positive;
(Readycult optional for E.coli: Kovac’s indole reagent added, immediate red ring, E. coli positive)
(465.360k9).
______
k. Enzyme tests not used to verify/confirm coliform on membrane filters or in broth cultures (465.360ik10).
______
l.
Media protected from light and only from commercially available source (465.360k2 k3).
______
QC m. Media and bottles checked for fluorescence before use (465.360k4,k6).
______
QC n. For Quanti-Tray, sealer checked monthly for leakage (465.360i15).
______
o. Reference comparators provided by manufacturer discarded by expiration date. (465.360i12).
______
p. Pre-incubation sample instructions followed:
Colilert-18 Quanti-Tray and Colisure: sample allowed to reach room temperature before incubation.
Colilert -18 P/A: sample pre-warmed to 35°C ± 0.5 C water bath 20 minutes or 44.5°C for 7-10 minutes.
Colilert P/A, Quanti-Tray, Readycult, Colitag: 24-hour incubation time includes time to bring sample
temperature to 35°C±0.5 C
Large sample load brought to room temperature before incubation (465.360k7).
______
8. Heterotrophic Plate Count (HPC)
SWTR (Count)
Pour Plate SM 22nd Ed 9215B
Heterotrophic
Plate Count
SimPlate Manufacturer’s Instructions
a. Work area disinfected.
______
b. No more than 1ml or less than 0.1ml of sample plated.
______
c. Refrigerated medium stored up to three months (465.360t4).
______
d. At least two replicate plates per dilution prepared for each sample (465.360t5).
______
e. Agar tempered to 44°C - 46°C before plating; melted agar used within three hours;
temperature control bottle containing media used (465.360t3).
______
f.
Center of media in containers not more than 2.5 cm from some surface (465.360t3).
______
g. Pipette tips not dragged across exposed ends of pipets in the pipet container or across lips and
necks of dilution bottles.
______
h. Pipettes not inserted more than 2.5 cm below surface of sample or dilution.
______
i.
When removing sample portions, pipette held at a 45° angle with the bottom of the tip touching the
inside neck of the sample or dilution bottle.
______
j.
When pipetting measured portions, the tip of the pipet rests on the inside bottom of the petri dish.
______
k. After flow has stopped, touched off once on a dry spot for 1 ml portions, if pipette is not a blow-out type;
remaining volume gently blown out from cotton-plugged blow-out-type pipette for 1 ml portions after
flow has stopped; 0.1ml portions not touched off.
______
l.
10ml -12 ml agar poured per plate (465.360t6).
______
m. Sample and agar mixed carefully (465.360t6).
______
n. Plates incubated in inverted position, no more than four high at 35°C ± 0.5°C for 48 ± 3 hours
(465.360t6).
______
18
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