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)

Conversion of a Biosafety Level 3 (BSL-3) lab to an Arthropod Containment Level 2 (ACL-2) lab

NITD @ Biopolis Singapore

NITD to move to Biopolis in April 2004 with official opening in July 2004

Nanos: Institute of Bioengineering

& Nanotechnology

(Proteos) Institute of Molecular and Cell Biology

Matrix: BioInformatics Institute

Genome Institute of Singapore

Helios Centros:

Bioprocessing Technology Centre

Chromos NITD

Min. of Education

Carpark ‘A’

Conversion of BSL-3 to ACL-2

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 3

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

Why ACL-2 (Arthropod Containment Level-2)

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 4

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.

ACL-2 lay-out

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 5

Ana

Animal room

Breeding room

Feeding room Dissection room

Infectious Area

Anteroom 1

Anteroom 2

High Temp & Humidity

Configuration of BSL3 HVAC system

6

Balancing Damper

AHU-8-2

AHU-8-1

BSL-3+/ABSL-3 Laboratory

Non- Return Damper

Isolation Damper

Non- Return Damper

•Isolation Damper

EF8-1 EF8-2

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only

Separate supply air only for the breeding room

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 7

Ana

Animal room

Breeding room

Feeding room Dissection room

Infectious Area

Anteroom 1

Anteroom 2

High Temp & Humidity

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

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 8

Different type of risks difference in Risk Assessment

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 9

Comparison of BSL-3 and ACL-2 (differences and similarities)

BSL-3 ACL-2 Room filter HEPA filter Air exhaust and supply covered with fine

mesh, including structures like sprinklers etc.

Room Pressure Negative Slightly negative

Waste disposal Decontamination tank and pass-through 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

How to contain a tiny mosquito- facility features

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 10

Ana Interlock

doors

Animal room

Breeding room

Feeding room Dissection room

Anteroom 1

Anteroom 2

Physical curtain

Sliding doors with alarm Sliding doors with alarm

Controlled access With air curtain

glovebox

Air curtain

Air curtain

Lower Ceiling

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!!!

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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

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Prevention of Mosquito Escapees contd.

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

| BSL3 to ACL2 | Jolly Qian | Oct 14 2015 | Biosafety | Business Use Only 13

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

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Insectary Operational Challenges contd.

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

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Novartis Institute For Tropical Diseases Management