Environmental. National Legionella Conference Sample Taking Workshop. Right Solutions Right Partner

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1 Environmental National Legionella Conference Sample Taking Workshop

2 Introductions Ashley Radbourne Senior Project and Field Services Manager Cassandra Hogan Business Development Officer Nathan King Lead Field Technician Please come and see us tomorrow at the ALS exhibition booth 2 National Legionella Conference Sample Taking Workshop

3 Outline What is a sample Why take a sample Water Risk Management Plan Stakeholder Consideration and Management Sample bottles Field Analysis Aseptic sampling Sampling for Legionella Sample points Sample integrity Submitting samples to the laboratory Legionella test and confirmation methods Interpreting results Additional analysis 3 National Legionella Conference Sample Taking Workshop

4 Legionella Monitoring What is a sample? 4 National Legionella Conference Sample Taking Workshop

5 Legionella Monitoring What is a sample? 1. A sample is a snapshot of the quality of the water in your system at the time of sampling. It is an indicator of the general health of your water system. A sample is a snapshot of the quality of the water in your system at the time of sampling It is an indicator of the general health of your water system 5 National Legionella Conference Sample Taking Workshop

6 Legionella Monitoring Why take a sample? Legionella is a commonly occurring bacteria found in both natural and artificial water systems Water temperatures within the range of C create conditions where Legionella is able to grow, with maximum proliferation in the range of C Legionella pneumophila has been indicated in Legionnaire s disease, which is a serious and potentially fatal form of pneumonia Those most at risk of infection are the immunocompromised Monitoring for Legionella can help identify and minimize the risk to immunocompromised patients Duty of care to occupants 6 National Legionella Conference Sample Taking Workshop

7 Legionella Monitoring Why take a sample? Sampling for Legionella is classified as verification monitoring enhealth 2015 guidelines state verification monitoring is used to confirm the quality of the water supply and determine whether existing control measures are effective. It is an important point to remember Sampling and analysis is for verification purposes only. It is a tool to help you assess whether the controls you have put in place to manage Legionella are effective It is not a safeguard against Legionella Do not sample for Legionella if you do not have a Water Risk Management plan in place. 7 National Legionella Conference Sample Taking Workshop

8 Legionella Monitoring Water Risk Management Plan Legislative Requirement Public Health Act 2005 and Public Health Regulation 2005 (both amended February 2017) Overarching guiding document 8 National Legionella Conference Sample Taking Workshop

9 Stakeholder Consideration and Management When sampling in healthcare facilities, you need to be aware of the requirements of that facility Sensitive areas include ICU, maternity, birth suites, mental health and palliative care Always ask the nurse in the area if it is ok to enter a room Be aware of protective clothing that may be required in specific areas e.g. scrubs, face masks, gowns, gloves, hair nets, booties If a room is not accessible, either finish sampling other areas and come back, or alternatively, sample the nearest available sampling point Be sensitive of the patients in the area, treat them with respect and do not tell them you are taking samples for Legionella testing Do not mention the L word 9 National Legionella Conference Sample Taking Workshop

10 Sampling Techniques for Potable Water - Bottles When taking samples for Bacteriological and Chemical tests, always take the Bacteriological sample first There are specific sample bottles for different types of analyses, they have different preservatives Using the incorrect bottle can interfere with your test results your laboratory will be able to provide the correct bottle types 10 National Legionella Conference Sample Taking Workshop

11 Sampling Techniques for Potable Water Field Analysis Field analyses can be performed at the time of sampling, tests include: Total Chlorine Free Chlorine Monochloramine ph Temperature Electrical conductivity Turbidity 11 National Legionella Conference Sample Taking Workshop

12 Sampling Techniques for Potable Water Aseptic Sampling Bacteriological samples must be taken aseptically This means to use techniques that prevent contamination of your sample with bacteria from the environment you are sampling in Tips for aseptic sampling: You must use a sterile container preferentially with Sodium Thiosulfate to neutralize chlorine residuals Wear gloves or wash/sanitise your hands Do not touch the inside of your sample bottle or sample bottle lid Do not put the bottle lid down on any surface when taking a sample Do not touch the sample point itself, only the tap handle Minimise the time your sample bottle is open to the environment 12 National Legionella Conference Sample Taking Workshop

13 Sampling Techniques for Legionella First Flush this is the first flow of water to come out of the tap this gives an indication of the water quality at the tap itself Timed flush times vary from, often 15 or 30 seconds, can be up to 2 minutes This gives an indication of the water quality of the water system/pipes Both types of samples can be taken at either cold or warm temperatures Most healthcare facilities have thermostatic mixing valves, which limit the temperature of the water to approximately 42 degrees 13 National Legionella Conference Sample Taking Workshop

14 Sampling Techniques for Legionella Sample Points Shower 14 National Legionella Conference Sample Taking Workshop

15 Sampling Techniques for Legionella Sample Points Basin 15 National Legionella Conference Sample Taking Workshop

16 Sampling Techniques for Legionella Sample Points Sensor Tap 16 National Legionella Conference Sample Taking Workshop

17 Sampling Techniques for Legionella Sample Points Zip Boiler 17 National Legionella Conference Sample Taking Workshop

18 Sampling Techniques for Legionella Sample Points Water Cooler 18 National Legionella Conference Sample Taking Workshop

19 Sampling Techniques for Legionella Sample Points Ice Machine 19 National Legionella Conference Sample Taking Workshop

20 Sampling Techniques for Legionella Sample Points Toilet 20 National Legionella Conference Sample Taking Workshop

21 Sampling Techniques for Legionella Sample Points Cooling Tower 21 National Legionella Conference Sample Taking Workshop

22 Sampling Techniques for Legionella Sample Points Surface Swab 22 National Legionella Conference Sample Taking Workshop

23 Sample Integrity Sample Details and HTs Sample details: Traceability and reference e.g. ID, time, date, any other relevant information Record on sample container and Chain of Custody (CoC) Sample holding time Recognised viability timeframe for target analyte 24 hours for bacteriological 23 National Legionella Conference Sample Taking Workshop

24 Sample Integrity Storage & Transport Sample storage and transport Chill but not freeze (preferably around 5 C per AS ) Preferentially use artificial ice bricks Warm and Cold samples should be transported separately Avoid possible secondary contamination from water ingress Secure transport 24 National Legionella Conference Sample Taking Workshop

25 Submitting a Sample to a Laboratory - COC 25 National Legionella Conference Sample Taking Workshop

26 Submitting a Sample to a Laboratory - COC Critical client information Turnaround time and traceability 26 National Legionella Conference Sample Taking Workshop

27 Submitting a Sample to a Laboratory - COC Sample details and required analysis Also any comments or special instructions 27 National Legionella Conference Sample Taking Workshop

28 Legionella Test Methods There are multiple methods for analysing Legionella Cultural Methods AS 3896, ISO and ISO Polymerase Chain Reaction PCR Rapid DNA Assay Field test kits The Public Health Regulation 2005 states that a prescribed test for Legionella Quantifies the number of legionella colony forming units (cfu) in the sample tested Is carried out by an accredited laboratory and is identified in the scope of the laboratory s accreditation Colony Forming Unit (CFU) 28 National Legionella Conference Sample Taking Workshop

29 Legionella Test Methods AS 3896:2017 Involves treating a water sample by several independent steps to optimise the recovery of Legionella species (direct, acid and heat treating) Samples are then spread onto selective culture media (agar) Incubated at 36 degrees for 7 days, plates examined at 4 and 7 days and then colonies present are counted and confirmed to a species level This method has a lower detection limit of 10CFU/mL This method has recently been reviewed and updated, you can no longer enumerate individual serogroups (SG1-SG14) Report now as Legionella pneumophila, Legionella species and Total Legionella (a sum of both) 29 National Legionella Conference Sample Taking Workshop

30 Legionella Test Methods ISO Acid treatment of a water sample to remove interfering bacteria and other species Filtered through a membrane, which is then placed on a culture media Incubated for 10 days (growth takes longer on a membrane then on culture media directly) Colonies counted and confirmed to a species level Standard reporting is to 1cfu/100mL, although up to 1L can be filtered giving a result of 0.1cfu/L 30 National Legionella Conference Sample Taking Workshop

31 Legionella Confirmation Methods Cultural method Include isolating a colony and subculturing (streaking) onto selective agar and incubating for a further 2-3 days, positive Legionella are determined, as they only grow on a certain type of agar Can then go through Immuno-agglutination (serotyping) for positive colonies to detect Legionella pneumophila serogroup 1 or serogroup This will be reported as detected or not detected. MALDI-TOF MS (Matrix assisted laser desorption ionization-time of flight mass spectrometry) Eliminates the 2-3 day confirmation time Uses Mass Spectrometry to measure a unique molecular fingerprint of an organism and is matched against a database with thousands of individual strains of micro-organisms Identifies individual species of Legionella Example of a confirmed Legionella spp. Mass Spectrum from MALDI- TOF MS 31 National Legionella Conference Sample Taking Workshop

32 Interpreting Your Results What do you do if you get a positive Legionella test? Don t Panic! Just because you have a positive Legionella result, does not mean you will have a Legionella outbreak Implement the controls in your Water Risk Management Plan Retest to ensure the controls have managed the risk 32 National Legionella Conference Sample Taking Workshop

33 Additional Laboratory Analysis Heterotrophic Plate Count (HPC) This analysis can give you an overall indication of the health of your water system and the effectiveness of the controls you have in place Pseudomonas aeruginosa Pseudomonas aeruginosa is a bacteria that is often found in biofilms. Analysis for this can help you assess if you have biofilm formation that could be harbouring Legionella bacteria E.coli Indicates faecal contamination of a water supply Turbidity Can impact efficacy of disinfection controls (e.g. UV) Sediment build up Metals Indicates potential leaching from within the system Disinfection by-products THMs, HAAs, Nitrosamines, etc. Safe Levels Governed by Australian Drinking Water Guidelines (ADWG) 33 National Legionella Conference Sample Taking Workshop

34 Questions and discussion Any Questions? 34 National Legionella Conference Sample Taking Workshop

35 Certificate of Attendance 35 National Legionella Conference Sample Taking Workshop