AS/NZS 3896:1998

1 - AS/NZS 3896:1998 WATERS-EXAMINATION FOR LEGIONELLAE INCLUDING LEGIONELLA PNEUMOPHILA
4 - PREFACE
5 - CONTENTS
6 - 1 SCOPE
6 - 2 REFERENCED DOCUMENTS
6 - 3 PRINCIPLE
6 - 4 SAFETY PRECAUTIONS
6 - 5 CULTURE MEDIA, REAGENTS AND REFERENCE CULTURES
6 - 5.1 Culture media
6 - 5.1.1 Buffered charcoal yeast extract agar with MWY selective supplement (MWY) or Buffered charcoal yeast extract agar...
7 - 5.1.2 Buffered charcoal yeast extract agar with BMPA selective supplement (BMPAa).
7 - 5.1.3 Buffered charcoal yeast extract agar (BCYEa).
7 - 5.1.4 Blood agar (BA).
7 - 5.1.5 Diluent: sterile 0.1% peptone or sterile quarter strength Ringer's solution.
7 - 5.2 Reagents
7 - 5.2.1 HCl/KCl buffer
7 - 5.2.2 Gram stain reagents
7 - 5.2.3 Latex agglutination reagents
7 - 5.2.4 Optional reagents for other identification tests
7 - 5.3 Reference organisms
7 - 5.3.1 Positive culture
7 - 5.3.2 Negative culture
7 - 5.3.3 Use of reference cultures
7 - 6 APPARATUS
7 - 6.1 Vortex mixer.
7 - 6.2 Water bath at 50 ±1°C.
7 - 6.3 Long-wave (360 ±20 nm) ultraviolet lamp.
7 - 6.4 Plate (or dissecting) microscope 10-20 × magnification.
7 - 6.5 Suitably calibrated micropipette capable of delivering the required volumes.
7 - 7 SAMPLE STORAGE
8 - 8 PROCEDURE
8 - 8.1 General
8 - 8.2 Isolation of Legionella-like organisms (LLOs)
8 - 8.2.1 Isolation procedure
8 - 8.2.2 Direct sample (untreated)
8 - 8.2.3 Heat-treated sample
8 - 8.2.4 Acid-treated sample
8 - 8.3 Incubation
8 - 8.4 Examination
9 - 8.5 Confirmation of legionellae
9 - 8.5.1 Procedure
9 - 8.5.2 Interpretation
9 - 8.6 Confirmation of L. pneumophila
10 - 8.7 Other tests for legionellae
10 - 9 CALCULATIONS
10 - 10 TEST REPORT
10 - 11 BIBLIOGRAPHY
12 - APPENDIX A - FLOW CHART FOR LEGIONELLAE TESTING
13 - APPENDIX B - CULTURE MEDIA
13 - B1 SCOPE
13 - B2 CHARCOAL YEAST EXTRACT AGAR BASE (CYE)
13 - B3 BUFFERED CHARCOAL YEAST EXTRACT AGAR (BCYEa)
13 - B3.1 Formulation
13 - B3.2 Growth supplement
13 - B4 BUFFERED CHARCOAL YEAST EXTRACT AGAR WITH BMPA SELECTIVE SUPPLEMENT (BMPAa)
13 - B4.1 Formulation
14 - B4.2 BMPA selective supplement
14 - B5 BUFFERED CHARCOAL YEAST EXTRACT AGAR WITH MWY SELECTIVE SUPPLEMENT (MWY)
14 - B5.1 Formulation
14 - B5.2 MWY selective supplement
14 - B6 BUFFERED CHARCOAL YEAST EXTRACT AGAR WITH GVPC SELECTIVE SUPPLEMENT (GVPC)
14 - B6.1 Formulation
14 - B6.2 GVPC selective supplement
15 - B7 TRYPTICASE SOYA + 5% BLOOD AGAR (BA)
15 - B8 PREPARATION OF ACID BUFFER
15 - B9 PEPTONE SOLUTION, 0.1%*
15 - B10 RINGER'S SOLUTION, FULL STRENGTH
15 - B10.1 Formulation
15 - B10.2 Ringer's solution, quarter strength
16 - APPENDIX C - FINAL READING SUMMARY
18 - AMENDMENT CONTROL SHEET