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Conversion of a Biosafety Level 3 (BSL-3) lab to an Arthropod Containment Level 2 (ACL-2) lab Jolly Qian, Biosafety/ACL-2 ops manager, PhD RBP Jason Khoo, Head of HSE and facilities Lakshmanan Ponnusamy, facility supervisor Novartis Institute for Tropical Diseases (NITD) NITD @ Biopolis Singapore (Proteos) Nanos: Institute of Institute of Molecular and Cell Biology Bioengineering & Nanotechnology Matrix: BioInformatics Institute Helios Genome Centros: Carpark ‘A’ Min. of Education Bioprocessing Technology Centre Institute of Singapore Chromos NITD NITD to move to Biopolis in April 2004 with official opening in July 2004 Conversion of BSL-3 to ACL-2 Decommission of BSL-3 (skip) Planning of an ACL-2 lab • Space- how many rooms do we need? • Conversion-how to deal with the M&E works? • Staffing-operation • Science Construction of the ACL-2 lab 3 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Why ACL-2 (Arthropod Containment Level-2) Support the Malaria drug discovery program Reference ‘ACME’ I. Arthropod Containment Guidelines by The American Committee of Medical Entomology of the American Society of Tropical Medicine and Hygiene II. Arthropod Containment Level 2 (ACL-2) must be practiced if working with exotic and indigenous arthropods infected with BSL-2 agents associated with animal and/or human disease, or that are suspected of being infected with such agents. 4 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only ACL-2 lay-out Animal room Feeding room Dissection room Infectious Area Anteroom Ana 2 Anteroom 1 HighBreeding Temproom & Humidity 5 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Configuration of BSL3 HVAC system Non- Return Damper Isolation Damper Balancing Damper AHU-8-2 NonReturn Damper EF8-2 EF8-1 AHU-8-1 •Isolation Damper BSL-3+/ABSL-3 Laboratory 6 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Separate supply air only for the breeding room Animal room Feeding room Dissection room Infectious Area Anteroom Ana 2 Anteroom 1 HighBreeding Temproom & Humidity 7 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Conversion Retained most of the BSL-3 infrastructures e.g. ACMV, gowning rooms, clean corridor area etc. etc. Except for the breeding room, which used a FCU to supply air, still using the old BSL-3 AHU/Exhaust->single pass, high ACH Bag-in Bag-out system × Brand new and white lab furniture, lighting system under benches Installed two humidifiers for the breeding room to back-up each other 8 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Comparison of BSL-3 and ACL-2 (differences and similarities) Different type of risks difference in Risk Assessment BSL-3 Room filter HEPA filter ACL-2 Air exhaust and supply covered with fine mesh, including structures like sprinklers etc. Room Pressure Negative Slightly negative Waste disposal Decontamination tank and passthrough autoclave A small stand-alone autoclave for solid waste and a ‘rice cooker’ for non-infectious liquid waste like larvae culture water Safety equipment BSC/Glovebox/ Respiratory protection program Glovebox (no ventilation!) and mosquito traps 9 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only How to contain a tiny mosquito- facility features Sliding doors with alarm Air curtain Feeding room Air curtain Animal room Sliding doors with alarm Dissection room glovebox Anteroom Ana 2 Interlock doors Anteroom 1 Breeding room Lower Ceiling Controlled access With air curtain Physical curtain 10 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Prevention of Mosquito Escapees Non-infected mosquito escapees • Not very unusual but it can be serious too • How and where does it happen? Look at the details in the procedure e.g. change glucose etc etc Infected mosquito escapees • Mosquito infection happens in strictly controlled environment so escapee should be very rare and serious case • Evacuation • Review each step in the infection and dissection process and plan in advance • Perform mock experiment with non-infected mosquitoes -be familiar with the procedure -build up confidence!!! 11 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Prevention of Mosquito Escapees contd. Regular check of mosquito traps it is good to have more than one trap for a relatively bigger room! Regular (quarterly) change of attractant in the traps Understand the mosquito life cycle 12 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Insectary Safety and Operational Challenges Staffing • Back-up system absolutely necessary! Weekend/holidays duties - directly impact the safety standard Working environment - Hot and humid - Low ceiling and no window 13 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Insectary Operational Challenges contd. Housekeeping is crucial for both of safety and scientific purpose!!! • No clutter • freeze the dirty trays or cages and then wash them immediately • mop floor frequently • regular change of lab coat 14 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only Thank for your attention! NITD insectary team and our collaborators (Duke-NUS and Environmental Health Institute): Andy Yip, Lim Liting, Pablo Bifani and Sudirman Made Ali Rusdiyah Novartis Institute For Tropical Diseases Management 15 | BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only