Elements of Biosafety Janet Peterson jpeterso@ 405-3975

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Elements of Biosafety Janet Peterson jpeterso@accmail.umd.edu 405-3975 Slide 2 BIOSAFETY: Preventing lab-acquired infections l Bacteria l Viruses l Fungi l Human blood, unfixed tissue l Human cell lines l Recombinant DNA Slide 3 Guidelines l NIH Guidelines for Experiments Involving Recombinant DNA Molecules Large Scale> 10 liters l Biosafety in Microbiological and Biomedical Laboratories-NIH/CDC l UM Biosafety Manual On DES webpage www.inform.umd.edu/des Slide 4 Regulations l OSHA Bloodborne Pathogens Standard l Maryland Waste Regulations l Shipping and packaging infectious substances DOT, UN, CDC, IATA Slide 5 Levels of Containment l BL1 - microorganisms that dont consistently cause disease in healthy adults E. coli K12, S. cerevisiae, polyomavirus Basic laboratory Standard Microbiological Practices Slide 6 Levels of Containment l BL2 - microorganisms of moderate potential hazard, transmitted by contact, ingestion, puncture Salmonella, herpesvirus, human blood Basic laboratory Standard Practices PLUS Slide 7 Levels of Containment l BL2 - Standard Microbiological Practices Plus: Training in handling pathogens Access to lab limited Extreme sharps precautions Use of BSC for aerosols Slide 8 Biosafety Cabinets Courtesy of the Baker Company Slide 9 Use of Biosafety Cabinet l Turn on fan 15 min before starting l Dont block grille l Disinfect work surface w/ 70% etoh l Discard pipets inside cabinet l Minimize movement of hands l Avoid use of flame unless necessary l Have cabinet certified annually Slide 10 Clean Bench l This is not a BSC l Air flows from back of cabinet, across work surface, and onto user. l This does not provide worker protection. Slide 11 Levels of Containment l BL3 - microorganisms that cause serious disease, transmitted by inhalation M. tuberculosis, yellow fever virus, hantavirus, Y. pestis (plague) Containment lab: double door entry; directional airflow; all work in biosafety cabinet Slide 12 Levels of Containment l BL4 - microorganisms that cause lethal disease, with no known treatment or vaccine Ebola virus, Marburg virus Maximum containment lab; positive pressure ventilated suits (moon suits) Slide 13 OSHA Bloodborne Pathogens Standard l Human blood, unfixed tissue, primary human cell culture, other potentially infectious materials l HIV,HBV, HCV Slide 14 OSHA Standard requires: l Annual training Web-based program/DES homepage l Free HBV vaccine l Use of Universal Precautions Slide 15 Universal Precautions l Treat ALL human blood and unfixed tissue as if it contains HIV and HBV Slide 16 Routes of Occupational Transmission l Puncture or cut (needlestick, contaminated broken glass) l Contact with broken skin l Splash to mucous membranes of eyes, nose, mouth Slide 17 Precautions for First Aid l Wear gloves l If conscious, have patient put pressure on wound l Use one-way valve for CPR Slide 18 Standard Microbiological Practices l NOT permitted in laboratories: Eating Drinking Smoking Handling contact lenses Pipetting by mouth Storing food and drink Slide 19 Standard Microbiological Practices l ALWAYS wash hands: After handling microorganisms and animals After removing gloves Before leaving laboratory Slide 20 Standard Microbiological Practices l Discard needles, razor blades, and scalpel blades into red, puncture resistant sharps containers l Dispose of broken glass into broken glass containers, never regular trash Slide 21 Standard Microbiological Practices l NEVER recap, bend, or break needles discard needles or sharps into biological waste bags discard needles into regular trash Slide 22 Standard Microbiological Practices l Decontaminate all biological waste (including BL1) before disposal Solid waste (Petri dishes, cultures): autoclave and put in dumpster Liquid waste: add disinfectant (bleach to 10%) and pour down drain Slide 23 Autoclaves l Autoclaves use pressurized steam to sterilize materials. l There is usually steam remaining in the chamber at the end of a liquid cycle. Slide 24 Autoclave Safety l Opening door at end of liquid cycle: Wear eye and face protection. Stand behind door when opening it. Slowly open door only a crack to allow residual steam to escape. Keep face away from door as it opens. Slide 25 Autoclave Safety l Removing liquids at end of cycle: Wait 5 min. before removing liquids. Liquids removed too soon may be super-heated and boil up and out of container. Aim mouth of flask away from face. Dont knock flask against bench. Slide 26 Standard Microbiological Practices l Decontaminate work surfaces daily and after any spill of viable material l Report accidents to the PI l Tell Health Care Provider that you work with infectious agents or chemicals Slide 27 Think Again l You've carefully thought out all the angles. l You've done it a thousand times. l Nothing could possibly go wrong, right ? Slide 28 Questions?