Best Practices for Decontaminating Biological Equipment & Spaces
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Do you have contamination in your biology lab? Preventing and remediating contamination in incubators, cold rooms, and biological safety cabinets allows your organization to run a smoother operation, reduce equipment downtime, and create a safe, environmentally compliant and healthy facility. Learn more at https://www.triumvirate.com/field-services/biological-decontamination-services.
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Best Practices for Decontaminating Biological Equipment & Spaces
- 1. Best Practices for Decontaminating Biological Equipment & Spaces Ryan Miller, Triumvirate Environmental
- 2. Meet Your Moderator James Ciccone
- 3. During this Webinar ✓ All lines will be muted – please communicate via the questions tab in your webinar panel. ✓ There will be a Q&A session at the end of the presentation – submit your question(s) anytime throughout the webinar. ✓ Slide deck and webinar recording will be sent to you tomorrow.
- 4. Meet Your Presenter Ryan Miller
- 5. Who are We? Focused within life sciences, healthcare, education and industrial markets, Triumvirate Environmental is a leading provider of enterprise waste management and EH&S services in North America. Service Categories Include: • Waste Management & Disposal • EHS Consulting Services • Field Services • Onsite Support Services www.triumvirate.com
- 6. Agenda Contamination Sources Biological Equipment and Spaces Case Studies Q&A
- 7. Poll Question When was the last time you had to decontaminate a room or piece of equipment? a. Within the past 2 years b. Within the past year c. Within the past 6 months d. Within the past month e. Never (that I’m aware of)
- 8. Contamination Sources Causes of contamination: • Laboratory personnel • Poorly developed or poorly executed procedures ▪ Unclean lab coats ▪ Mislabeling ▪ Cross contamination ▪ Poor cell culture management • Unclean/dirty work environment • Poorly designed spaces
- 9. Mycoplasma • Smallest free-living organism • Considered to be the simplest bacteria • Lacks a cell wall ▪ Resistant to common antibiotics • Must function as parasites to meet energy and biosynthesis demands • Size ~100 nm undetectable by optical microscopy • Multiply within cells
- 10. • Ubiquitous • Broad range of hosts ▪ Humans and other mammals ▪ Birds ▪ Reptiles ▪ Fish ▪ Insects ▪ Plants Mycoplasma
- 11. • First reported in cell culture in 1956 ▪ Continues to challenge scientists • No perceivable reduction of cell culture infection rates ▪ Insufficient protection ▪ Carelessness Mycoplasma
- 12. Mycoplasma in Cell Cultures • Compete with host cells for nutrients • Can alter DNA, RNA, and protein synthesis • Diminishes amino acid and ATP levels • Introduces chromosomal alterations • Modify host-cell plasma membrane antigens
- 13. Sources of Mycoplasma in Cell Cultures • Infected cells sent from another lab • Contaminated cell culture medium reagents • Laboratory personnel infected with M. orale or M. fermentans • Contamination can spread rapidly to other cell lines
- 14. Mycoplasma Disinfection • Cell Culture ▪ Despite resistances, addition of suitable antibiotics is best (Drexler & Uphoff, 2003) ➢Quinolones, tetracyclines, and macrolides • Surfaces ▪ Autoclave – compatible materials, fully efficient ▪ 70% ethanol – fully efficient, complete inactivation after 5 min. exposure ▪ Alkaline cleaner – increased efficiency with increased concentration, complete inactivation at 8% ▪ Chlorine Dioxide or Vaporized Hydrogen Peroxide– very efficient for enclosures or small rooms ▪ Quaternary ammonium compounds – poor efficiency
- 15. Agenda Contamination Sources Biological Equipment and Spaces Case Studies Q&A
- 16. Incubators
- 17. Incubator Contamination • Fans become loaded with debris • Buildup occurs on sensors and gaskets • Basic surface cleaning & gas disinfection does not remove this buildup • Leads to contaminated media
- 18. • Procedure is make and model specific • Follow methods recommended by manufacturer • Manufacturer provides replacement parts kit (filters, gaskets, plugs, etc.) Incubator Contamination
- 19. • Clean and/or replace removed parts including: ▪ Shelves ▪ Shelf supports ▪ Plugs & grommets ▪ Ducts ▪ Humidity pan ▪ Gaskets ▪ Filters ▪ Fans Incubator Contamination
- 20. Preventative Maintenance Make and model specific, an example from Thermo Fisher 3110
- 22. BSC Contamination • Internal parts and HEPA filter become loaded with debris • Basic cleaning does not remove this buildup • Leads to contaminated media
- 23. When is BSC Decontamination Needed? • After large spills of chemicals or biological materials • Prior to any maintenance work, filter changes, and performance tests that requires access to any contaminated portion of the BSC • Decommissioning, disposal, or salvage • Initiating a new work program or production run • Relocation from one room/building to another
- 24. • BSCs decontaminated per the National Sanitation Foundation (NSF)/American National Standards Institute (ANSI) 49 Standard for Biological Safety Cabinet Decontamination • Includes removing panels and removal of bulk materials and debris • BSC is sealed and a Mini Chlorine Dioxide System (MCS) is utilized to disinfect the cabinets BSC Contamination
- 25. Mini Chlorine Dioxide System • Utilizes chlorine dioxide (CD) gas for biological decontamination per NSF/ANSI 49 - 2016 • Fast and safe device for decontamination of BSCs • Built in charcoal gas-scrubbing unit allows technician to remove CD without external venting
- 27. CD Gas and VHP Attributes
- 28. Poll Question Which decontamination technologies do you typically use? (select all that apply) a. VHP b. ClO2 c. Other d. Both
- 29. Cold Rooms
- 31. • Mold needs moisture • Condensation can lead to mold growth Cold Room Contamination
- 32. Mold Remediation • Eliminate moisture sources • Remove all cardboard and porous materials • Use a biological disinfectant (bleach) to clean all surfaces • Run HEPA filtered vacuum to capture airborne spores
- 33. Confirmation • Spore trap analysis • Surface wipe samples
- 34. Routine Maintenance • Prevent contamination by being proactive • Setup a routine: ▪ Full decontamination bi-annually ▪ Full decontamination between operational changes ▪ Full decontamination if contamination is identified
- 35. Poll Question How often do you do routine contamination prevention of your cold rooms? a. Never b. Every 1-6 months c. Every 6-12 months d. Every 12-24 months
- 36. Agenda Contamination Sources Biological Equipment and Spaces Case Studies Q&A
- 38. • Routine maintenance only included incubator “self-clean” • Buildup in the fan housing, underneath baffles, and behind sensors • Full decontamination and consumable replacement mitigated the issue Incubators Contaminating Media
- 43. Cold Room Mold Contamination
- 44. Cold Room Mold Contamination
- 45. Agenda Contamination Sources Biological Equipment and Spaces Case Studies Q&A
- 46. Summary • Establish a preventative maintenance schedule • Perform routine decontaminations • Facilitate training and teaching • Determine appropriate strategies to minimize or manage the risk of contamination • Communication is the key
- 47. Questions?
- 48. THANK YOU! Ryan Miller: rmiller@triumvirate.com Engineering Project Manager Triumvirate Environmental Call Us! 1-888-834-9697 www.triumvirate.com