AS 3896:2017 pdf free.Waters – Examination for Legionella spp – including Legionella pneumophila.
This Standard sets out a method for isolating and estimating the number of Legionelia pneumophila and a range of other Legionella spp. in water. This method is applicable to all water samples. including recreational, industrial, waste and natural waters.
NOTES:
I A flow chart of the procedure is shown in Appcndix A.
2 While this method is suitable for the testing of all waters, there is a separate method (AS 5132) that is designed specifically for concentration of water where a lower limit of detection is required due to public health concerns.
3 This method will isolate the species L. pneuniophi!a but not all other Legionella spp. L. pneuinophlla scrogroup 1 accounts for the majority of lcgionellae infections.
4 Conditions that favour the isolation of L. pneu,nophila do not necessarily apply to some
Legionella spp. However, treatments and media included in this method are designed to
enhance the recovery of other species in addition to L. pneuinophila.
3 PRINCIPLE
The sample of water is cultured on specified media directly as well as after heat and acid treatments. Presumptive Legioneilci colonies are taken through con firmation tests. Confirmed Legionella isolates can be speciated and serotyped as required.
4 SAFETY PRECAUTIONS
The safety precautions for microbiological laboratories, as described in AS/NZS 2243.3, shall be observed.
Legionella cultures on solid culture media can be handled safely by trained staff on the open bench in a conventional microbiology laboratory conforming to Containment Level 2. Infection is caused by inhalation of the microorganism and it is advisable therefore to assess all procedures for their ability to produce aerosols. If in doubt, carry out the work in a biological safety cabinet.
5 CULTURE MEDIA, REAGENTS AND REFERENCE CULTURES
5.1 Culture media
The culture media shall be as follows:
(a) Buffered charcoal yeast extract agar (BCYE).
(b) Buffered charcoal yeast extract agar with MWY selective supplement (MWY) or buffered charcoal yeast extract agar with GVPC selective supplement (GVPC).
(c) Buffered charcoal yeast extract agar with BMPA selective supplement (BMPA) or buffered charcoal yeast extract agar with selective supplements (BCYE+AB).
(d) Buffered charcoal yeast extract medium without L-cysteinc (BCYE-Cys) or Blood Agar (BA).
(e) Diluent: sterile 0.1% peptone or sterile quarter strength Ringer’s solution.
NOTE: See Appendix B.