Recreational Water Facilities and Public Pools Spas

Specimen Collection and Handling

Specimen Requirements

Test Requested Required Requisition(s) Specimen Type Minimum Volume Collection Kit

Total Coliform, E. coli, P. aeruginosa, S. aureus and HPC

Water collected from a public recreational water facility

200 mL (to the fill line on the bottle)

PHO laboratory water collection bottle

Submission and Collection Notes

1

Complete the required information on the requisition (i.e. submitting agency name including the sub office if this office submitted the sample, date and time collected and sample type). The sample will not be tested if all of the required information is not complete or accurate when received at the laboratory.

2

Remove one barcode from the bottle and apply it to the top copy of the requisition in the “Barcode” field. Remove a second barcode and apply it in the corresponding location on the second copy of the form. Samples will not be processed if the barcodes are not affixed to the requisition. If the barcode is not able to be removed, write the barcode number in the “Barcode” box on the form.

3

Examine the lid of the bottle. If the tamper evident ring has separated from the cap use another bottle.

4

Remove cap and ensure it is kept away from sources of contamination.

  • Do not touch the inside of the lid.
  • Do not put the lid down.
  • Do not ingest the sodium thiosulfate in the bottle. For accidental exposure get medical attention.
5

Hold the bottle near the base and plunge the bottle, neck downward, at a 45 o angle to 30 cm and collect water within one metre of the water surface. Fill the bottle at that depth with a sweeping forward motion ahead of the hand, bringing the bottle up toward the water’s surface.

  • Take samples in an appropriate location of the pool (e.g. if equipped with a filter, collect from sampling ports provided in the filter’s return and discharge lines) in order to provide an accurate representation of the water in the pool.  For pools equipped with a filter, samples may be collected from sampling ports provided in the filter’s return and discharge lines.
  • Keep the bottle away from your other hand and body parts.
  • Avoid contamination from floating debris.
6

Fill to the 200 mL fill line.

  • Do not overfill.
  • If overfilled, remove some of the water so the bottle is filled to 200 mL line.
7

Tighten the cap on the collection bottle. Leaking samples will not be processed; a new sample and requisition will be required.

Timing of Specimen Collection

Ensure adequate time to collect and transport the sample to the laboratory. It must be tested within one calendar day of collection.

Limitations

Samples will not be tested if the acceptance criteria are not met. Refer to the Public Health Inspector’s Guide to Environmental Microbiology Laboratory Testing.

Storage and Transport

Retain a copy of the completed requisition that includes the barcodes. Specimens should be stored at 2-8 °C following collection and shipped to PHO laboratory inside insulated containers or in a cooler. Pack with frozen ice packs. Do not freeze samples. Samples must be shipped within 24 hours of collection and tested within one calendar day.

Special Instructions

This test is only offered for approved Official Agency clients (i.e., public health unit or Ministry of Natural Resources and Forestry Ontario Park).

Instructions For Official Agencies Submitting Water Samples to the Public Health Ontario Laboratory provides step by step instructions for completing the documentation and submitting/relinquishing the sample. For legal samples the chain of custody must be complete when the sample is received at the laboratory (Refer to Instructions For Official Agencies Submitting Water Samples to the Public Health Ontario Laboratory).

Requisitions and Kit Ordering

Test Frequency and Turnaround Time (TAT)

Samples are routinely accepted at the laboratory Monday – Thursday except holidays during regular operating hours. For submission around statutory holidays, refer to PHO laboratory holiday schedule.

Please contact the PHO laboratory Customer Service Centre prior to the submission of samples that will be received outside of these hours.

The TAT is up to 4 days for Total Coliform and E. coli, up to 5 days for HPC and P. aeruginosa and up to 6 days for S. aureus

STAT and Critical Samples Testing

Please contact the PHO laboratory Customer Service Centre prior to the submission of samples that will be received outside of regular operating hours or the PHO laboratory Duty Officer at (416) 605-3113 if outside of Customer Service's hours. Samples of an urgent nature (e.g. STAT) will be processed and read with no undue delay.

Test Methods

Specimens are tested for the microbial indicators: 

  • Total Coliform and Escherichia coli (E. coli) (method modified from MECP E3407: Membrane Filtration Method Using DC Agar for the Simultaneous Detection and Enumeration of Total Coliforms and Escherichia coli in Drinking Water).
  • Staphylococcus species (including S. aureus) (method modified from Standard Methods for the Examination of Water and Wastewater 9213B) and Pseudomonas aeruginosa (method modified from Standard Methods for the Examination of Water and Wastewater 9213 E. Membrane Filter Technique for Pseudomonas aeruginosa). 
  • The Heterotrophic Plate Count (HPC) test is performed using the Spread Plate method (modified from MECP E3408: The Spread Plate Method for the Enumeration of Aerobic, Heterotrophic Bacteria in Drinking Water).

By submitting a sample with a PHO Laboratory requisition, the submitter accepts Public Health Ontario’s methodology.

Reporting

Results are reported to the submitter as indicated on the requisition.

Additional Information

Microbiological sampling may be conducted to evaluate and monitor cleaning and sanitizing procedures and to ensure that a health hazard does not exist. Refer to the Ministry of Health and Long-Term Care’s Recreational Water Protocol and Operational Approaches for Recreational Water Guideline

Mis à jour le 20 juill. 2020