Presentation on theme: "Elements of Biosafety Janet Peterson 405-3975."— Presentation transcript:
Elements of Biosafety Janet Peterson
BIOSAFETY: Preventing lab-acquired infections l Bacteria l Viruses l Fungi l Human blood, unfixed tissue l Human cell lines l Recombinant DNA
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
Regulations l OSHA Bloodborne Pathogens Standard l Maryland Waste Regulations l Shipping and packaging infectious substances –DOT, UN, CDC, IATA
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
Levels of Containment l BL2 - microorganisms of moderate potential hazard, transmitted by contact, ingestion, puncture – Salmonella, herpesvirus, human blood –Basic laboratory –Standard Practices PLUS
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
Biosafety Cabinets Courtesy of the Baker Company
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
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.
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
Levels of Containment l BL4 - microorganisms that cause lethal disease, with no known treatment or vaccine è Ebola virus, Marburg virus è Maximum containment lab; positive pressure ventilated suits (moon suits)
OSHA Bloodborne Pathogens Standard l Human blood, unfixed tissue, primary human cell culture, other potentially infectious materials l HIV,HBV, HCV
OSHA Standard requires: l Annual training –Web-based program/DES homepage l Free HBV vaccine l Use of Universal Precautions
Universal Precautions l Treat ALL human blood and unfixed tissue as if it contains HIV and HBV
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
Precautions for First Aid l Wear gloves l If conscious, have patient put pressure on wound l Use one-way valve for CPR
Standard Microbiological Practices l NOT permitted in laboratories: è Eating è Drinking è Smoking è Handling contact lenses è Pipetting by mouth è Storing food and drink
Standard Microbiological Practices l ALWAYS wash hands: è After handling microorganisms and animals è After removing gloves è Before leaving laboratory
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
Standard Microbiological Practices l NEVER –recap, bend, or break needles – discard needles or sharps into biological waste bags –discard needles into regular trash
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
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.
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.
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.
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
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 ?