Presentation on theme: "Janet Peterson email@example.com 405-3975 Elements of Biosafety Janet Peterson firstname.lastname@example.org 405-3975."— Presentation transcript:
1 Janet Peterson email@example.com 405-3975 Elements of BiosafetyJanet Peterson
2 BIOSAFETY: Preventing lab-acquired infections BacteriaVirusesFungiHuman blood, unfixed tissueHuman cell linesRecombinant DNABiosafety is the section of Environmental Safety that addresses working safely in the laboratory with microorganisms and rDNA (organisms whose genetic material has been altered).Its goal is to help prevent lab-acquired infections while allowing researchers to work with organisms of various levels of hazard. This is done by advising researchers of the containment practices that are appropriate for the organisms they are handling, based on the level of risk presented by the organism.1
3 GuidelinesNIH Guidelines for Experiments Involving Recombinant DNA MoleculesLarge Scale > 10 litersBiosafety in Microbiological and Biomedical Laboratories-NIH/CDCUM Biosafety ManualOn DES webpageVarious regulations and guidelines have been developed to protect workers and the environment. They define work practices required to work safely with biological material of all levels of hazard. All Principal Investigators must register their experiments that involve the use of rDNA, infectious microorganisms, and human blood, unfixed human tissue, and primary human cell lines with DES.2
4 Regulations OSHA Bloodborne Pathogens Standard Maryland Waste RegulationsShipping and packaging infectious substancesDOT, UN, CDC, IATA
5 Levels of ContainmentBL1 - microorganisms that don’t consistently cause disease in healthy adultsE. coli K12, S. cerevisiae, polyomavirusBasic laboratoryStandard Microbiological PracticesThere are 4 levels of biological containment that allow laboratory researchers to work safely with infectious agents of various levels of risk. They are designed to prevent lab-associated infections.BL1 is the lowest level of containment. At UM there are many BL1 labs, most of which are molecular biology labs that use non-pathogenic strains of E. coli for cloning..3
6 Levels of ContainmentBL2 - microorganisms of moderate potential hazard, transmitted by contact, ingestion, punctureSalmonella, herpesvirus, human bloodBasic laboratoryStandard Practices PLUSBL2 is used for infectious agents that are transmitted by contact, ingestion, or puncture with contaminated sharp object. There are several BL2 labs at UM.4
7 Levels of Containment BL2 - Standard Microbiological Practices Plus: Training in handling pathogensAccess to lab limitedExtreme sharps precautionsUse of BSC for aerosolsBL2 labs are similar to BL1 laboratories, with the addition of these specific practices.
8 Biosafety Cabinets Courtesy of the Baker Company
9 Use of Biosafety Cabinet Turn on fan 15 min before startingDon’t block grilleDisinfect work surface w/ 70% etohDiscard pipets inside cabinetMinimize movement of handsAvoid use of flame unless necessaryHave cabinet certified annually
10 Clean Bench This is not a BSC Air flows from back of cabinet, across work surface, and onto user.This does not provide worker protection.
11 Levels of ContainmentBL3 - microorganisms that cause serious disease, transmitted by inhalationM. tuberculosis, yellow fever virus, hantavirus, Y. pestis (plague)Containment lab: double door entry; directional airflow; all work in biosafety cabinetBL3 containment is used for work with infectious agents that are transmitted by inhalation. We have 2 BL3 labs at UM, but neither is currently in use.7
12 Levels of ContainmentBL4 - microorganisms that cause lethal disease, with no known treatment or vaccineEbola virus, Marburg virusMaximum containment lab; positive pressure ventilated suits (moon suits)There are only a very few BL4 labs in the country: CDC in Atlanta; NIH and Ft. Detrick in MD. BL4 is used for work with the most hazardous microorganisms. Hot Zone described BL4 containment.8
13 OSHA Bloodborne Pathogens Standard Human blood, unfixed tissue, primary human cell culture, other potentially infectious materialsHIV,HBV, HCVThe OSHA BBP standard was published in 1991 to address concerns over occupational transmission of HIV to healthcare workers.BBPs are microorganisms present in the blood of infected persons, which are transmitted by blood-to-blood contact. Not casual contact or by inhalation.If you work with human material, you are covered by the BBP Standard.
14 OSHA Standard requires: Annual trainingWeb-based program/DES homepageFree HBV vaccineUse of Universal PrecautionsIf you work with human material, OSHA requires:annual training of employees. If you work with human material, please take the interactive web based training available on our web page.provision of HBV vaccine by employer at no cost to employee;use of Universal Precautions when handling human materialtreating ALL material as if it may contain BBPs
15 Universal Precautions Treat ALL human blood and unfixed tissue as if it contains HIV and HBVEven if material has been tested for BBPs such as HIV and HBV, it is possible that the test was performed before a detectable level of antibodies were produced, and the virus may be present. Therefore, ALL human material should be treated as infectious. Even if it is from your lab partner, because you can’t tell if someone is infected.If you work with human material, please take the interactive web based training available on our web page. (Copy of page in handout).5
16 Routes of Occupational Transmission Puncture or cut (needlestick, contaminated broken glass)Contact with broken skinSplash to mucous membranes of eyes, nose, mouthRoutes of occupational transmission of BBP.Optional:Routes of transmission in the general population:Sexual, both homosexual and heterosexualSharing needles among injecting drug users.Blood transfusions prior to screening of blood supply (1980s??).Mother to unborn child.6
17 Precautions for First Aid Wear glovesIf conscious, have patient put pressure on woundUse one-way valve for CPR
18 Standard Microbiological Practices NOT permitted in laboratories:EatingDrinkingSmokingHandling contact lensesPipetting by mouthStoring food and drinkThese are the basic practices when working with any microorganisms, and are the foundation for BL1 containment. They are designed to prevent transmission by contact, ingestion, and puncture.These practices are appropriate for ALL laboratories that work with radioisotopes and chemicals as well as microorganisms.9
19 Standard Microbiological Practices ALWAYS wash hands:After handling microorganisms and animalsAfter removing glovesBefore leaving laboratoryGloves are not a substitute for hand washing.10
20 Standard Microbiological Practices Discard needles, razor blades, and scalpel blades into red, puncture resistant sharps containersDispose of broken glass into “broken glass” containers, never regular trashIn your handouts there is an information sheet on handling sharps safely.Sharps containers are available from Chemistry Stores and scientific distributors such as Fisher and VWR.Sharps containers are usually red, puncture-resistant plastic. Do not overfill sharps containers. Please call DES for pick-up of filled sharps containers. They do not need to be autoclaved before pick-up.11
21 Standard Microbiological Practices NEVERrecap, bend, or break needlesdiscard needles or sharps into biological waste bagsdiscard needles into regular trashMost needle sticks occur when recapping needles. If you must recap the needle, use a one-handed technique.If you get stuck by a needle, wash the area with soap and water, then go to the University Health Center for follow-up as soon as possible. Anti-retroviral drugs are more effective if given within 2-3 hours after needle stick.12
22 Standard Microbiological Practices Decontaminate all biological waste (including BL1) before disposalSolid waste (Petri dishes, cultures): autoclave and put in dumpsterLiquid waste: add disinfectant (bleach to 10%) and pour down drainUse only autoclavable bags in autoclave. Put bags of waste in stainless steel or polypropylene tray during autoclaving to contain any leaks.There is information on autoclave safety and Biological Waste Disposal in the handout.13
23 Autoclaves Autoclaves use pressurized steam to sterilize materials. There is usually steam remaining in the chamber at the end of a liquid cycle.
24 Autoclave Safety 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.
25 Autoclave Safety 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.Don’t knock flask against bench.
26 Standard Microbiological Practices Decontaminate work surfaces daily and after any spill of viable materialReport accidents to the PITell Health Care Provider that you work with infectious agents or chemicalsInform your healthcare provided that you work in a laboratory, and describe the types of hazardous materials you handle.14
27 Think Again You've carefully thought out all the angles. You've done it a thousand times.Nothing could possibly go wrong, right ?