1. Semi – Rigid Isolator Units Charles River Laboratories Consulting & Staffing Services

2. Terminology Flexible Film SRI HEPA Port Gasket Magnahelic Gauge

3. Isolator History & Function Used since 1950’s Flexible Film or Rigid contruction Biocontainment  Negative Air Pressure Bioexclusion  Positive Air Pressure  “Biosecurity” at Charles River

4. Features Typical forward facing port Gasket will change color when sealed properly Gasket Clamp Hinge

5. Features Flexible Front HEPA Filter Glove Arm External Filter

6. Isolator Supplies Nitrile Gloves Electrical Tape O-Rings

7. Leak Detection Snoop® Apply liquid to area suspected of leak and watch for bubbles Alternative  Mild Soap solution

8. Air flow Negative Positive

9. Keeping it Clean Wescodyne® Clidox  Bleach Solution Windex®/Cinch® Peracetic Acid Chemical Incompatibility  Check MSDS

10. Ergonomics Your back should always be as straight as possible. Hydraulic tables allow the user to adjust the height!

11. Filter Change Filters should be checked daily Filters need to be changed when the magnahelic gauge increases by 0.05 (i.e. from 1.0 to 1.5)

12. Filter Pop Quiz Which filter assembly needs to be replaced?

13. Testing for deterioration This rim should appear green even when stretched. If small holes or cracks are present glove must be replaced. Punctures from animal bites or instruments also warrant glove replacement.

14. Glove and flexible surface checks Check the magnahelic gauge Pull the gloves inside out, pulling on the cuff only Check for holes, punctures, tears, or stretched areas Then recheck the magnahelic for proper pressure Wipe down the outside of the flexible front surface Semi-Rigid Isolator Daily Checks and Inspections

15. Magnahelic and Ball Valve Ball Valve Magnahelic Gauge Fan Motor

16. Magnehelic gauge check Check gauge and record twice daily Gauge reads what the pressure is inside the isolator (+.10 or -.10) Reads in Inches of Water Not reading proper pressure report immediately DO NOT adjust equipment unless properly trained Semi-Rigid Isolator Daily Checks and Inspections

17. Filter Locations HEPA filter External Filter

18. But how do you use it?

19. Introduction of supplies

20. Supplies always in unit Disinfectants Sharps Forceps Scraper Marking Pens Absorbent Towels Paper Bags Hand Broom

21. Supplies introduced to unit Feed Bedding Cages Water Sterilized vs Irradiated

23. Port Door

24. Loading the Isolator: Remove all jewelry, watches and especially rings Don protective glove liners Gather appropriate supplies Open the outside port door Place supplies into port –Spray down supplies with disinfectant/sterilent Allow sufficient contact time 20 – 40 minutes Secure outer door How to Use the Semi-Rigid Isolator Units

25. Loading the Isolator: continued Push the sleeves into the isolator by holding onto cuff Over stretching the gloves will cause damage Open the port door Remove items from port into the isolator After all items have been removed, shut and lock port door clamps How to Use the Semi-Rigid Isolator Units

26. Unloading the Isolator: Once everything is ready to be removed from the isolator spray the inside of the port completely Spray all items before placing them into port Once all items are sprayed down with Clidox the port can be loaded After everything is loaded the port is then closed Set the timer according to SOP/Chemical Manufacturer How to Use the Semi-Rigid Isolator Units

27. Unloading the Isolator: When the timer goes off you may unload the port Contaminated items should be disposed/sanitized appropriately. How to Use the Semi-Rigid Isolator Units

28. Label door with time initials Set exposure timer Document event in log

29. Introducing/Removing Animals to Isolator Supplies  Biosafety Cabinet  Plastic bottle/containers Nalgene®  Disinfectant  Forceps

30. Introducing/Removing Animals to Isolator Transfer into bottle/container inside BSC  Leave lids unsealed until all containers are filled Use same introduction procedures as supplies Ensure animals are promptly removed from bottle/container  Air supply in sealed container must be established

31. Removing Animals from Isolator Return to plastic bottles/containers Double wrap in bag  Paper or plastic? Use regular “spray out” procedures Air supply for animals limited!

32. Inside of the isolator check Shelving and all surfaces for cracks or cuts in the plastic Light bulb and casing Port and clamps to ensure that it seals inside and out If any one of these items are wrong or not working report immediately DO NOT leave trash inside the isolator DO NOT leave the isolator dirty Animal checks inside the isolators Check feed and water levels in each cage Observe the animals to ensure health status Check for and change any wet cages Semi-Rigid Isolator Daily Checks and Inspections

33. Cage changing Before cage changing begins ensure that all of the daily checks have been completed Make sure all items for changing are inside the isolator before starting, this will help with being efficient Change cages After the cages have been changed remove all trash and dirty bedding from the isolator Make sure all animals are present and safely in their home cages Wash down the inside of the isolator, all surfaces including the inside of the glove sleeves Once all trash is removed wipe down the port Filter checks Check the outside filter for any dirt, debris, or damage Report to a supervisor if something is found Semi-Rigid Isolator weekly Checks and Inspections