Safety Highlights Lab Safety Biosafety Chemical Safety Radiation

Safety Highlights Lab Safety Biosafety Chemical Safety Radiation Safety

Highlights Training Requirements n Resources n Responsibilities n Lab Safety Plan n – Dept. ME Safety Plan n. U of MN Safety Plan

Training Requirements All ME employees n Initial training : 2 options to satisfy – In person, presented by EHS 1. Lab Safety and Hazardous Waste Training – Online 1. 2. 3. n Intro to Research Safety Chemical Waste Mgmt Annual refresher training – By DEHS (in the fall) or ME

Training Requirements Lab members Procedure- or lab-specific training requirements n Animal handling – – – n IACUC Certification (SPF Training) (Controlled substances training) Human or NHP tissue exposure – Bloodborne pathogens (Online) – (NHP Training) Cont.

Training Requirements Lab members (cont. ) n Laser – Contact Rad Safety, EHS n n n Training Set up Working with radioactive materials – Radioactive materials n Radiation Safety Training

Training and Safety Resources Dept. EHS n DEHS: Training http: //www. dehs. umn. edu/training/ – Online training – Registration for in person training n DEHS: Assistance and resources – 6 -6002

Training and Safety Resources Dept. Mechanical Engineering n Dept. Mechanical Engineering: Safety Web page https: //me. umn. edu/info/services/safety/index. s html Index from ME Safety page: – Emergency Information and Phone Numbers (pdf) – Department Lab Safety Plan (pdf) (links to appropriate resources are included in this plan) – Annual Update Attendance Form (pdf) – Initial Update Attendance Form (pdf) – Teaching Lab Safety Form (pdf) – Laboratory Audit Checklist (pdf)

Training and Safety Resources Bioheat and Mass Transfer Lab n Laboratory Safety Handbook (on bookshelf) n n Chemical Waste handling and packaging instructions Compressed gas safety information Annual departmental training notes Transition to online Lab Safety Handbook – Possible excepted items saved as hardcopy, as required by U of MN Safety Plan

Responsibilities: PI / Supervisors/Principal Investigators n The immediate supervisor of a laboratory employee is responsible for: 1) assuring that potential hazards of specific projects have been identified and addressed before work is started; 2) ensuring there are written, laboratory-specific standard operating procedures for the protocols carried out in the laboratory that incorporate directions about how to mitigate the hazards of the procedures. 3) informing and training employees regarding the specific hazards in their area and in the work they will be doing; 4) scheduling time for the employee to attend designated training sessions; 5) enforcing U of MN safety policies and safe work practices; 6) conducting periodic audits of the research space under the supervisors control; 7) reporting hazardous conditions to the college or departmental research safety officer; 8) investigate laboratory accidents and send an Accident Investigation Worksheet (Appendix N) with recommendations to the departmental research safety officer for review. Performance will be measured by: • home department's documentation of current, pertinent safety training for the supervisor and each employee in the supervisor's group; • home department's documentation of regular audits for laboratory space under the control of the supervisor. (from Dept. ME Safety Plan)

Responsibilities: Employee n Employees who have significant responsibility for directing their own laboratory work are – responsible for assuring that potential hazards of specific projects have been identified and addressed before work is started. n n n All laboratory employees however, are responsible for: • attending safety training sessions; • following safety guidelines applicable to the procedures being carried out; • assuring that required safety precautions are in place before work is started; and • reporting hazardous conditions as they are discovered. (from Dept. ME Safety Plan)

U of MN Safety Plan n n n n Chapter 5 Working with Chemicals Introduction Prudent Planning General Procedures for Working with Hazardous Chemicals Working with Substances of High Toxicity Working with Biohazardous and Radioactive Materials Working with Flammable Chemicals Working with Highly Reactive or Explosive Chemicals Working with Compressed Gases Chapter 6 Working with Laboratory Equipment Introduction Working with Water-Cooled Equipment Working with Electrically Powered Laboratory Equipment Working with Compressed Gases Working with High/Low Pressures and Temperatures Using Personal Protective, Safety, and Emergency Equipment Emergency Procedures

Safety Plan and SOPs for ‘BMTL’ Safety Plan for ‘BMTL’ n n – – – n n n – – – Training plan Each lab member be trained according to Dept. ME standards, and, Each lab member be trained according to specific research uses Training records Record of lab members’ training – tracking list Hardcopy documentation of training – in Lab Safety Handbook (currently) Transition to softcopy documentation via BMTL web site Lab safety equipment Fire extinguisher, eye wash and shower Hazards and restrictions posted Biohazardous culture or waste Sharps Radioactivity Lab audit (internal) every 6 months Will develop an audit procedure and tracking

Safety Plan and SOPs for ‘BMTL’ (cont. ) n Each individual should identify hazards associated with any research procedure, then: 1. 2. Include identification of hazards in SOP (in section on Safety) Include instructions for “mitigation of hazard” in SOP (in section on Safety) 1. 2. 3. 4. Handling of materials Labeling of materials or waste Instructions for waste disposal Post hazard areas, if nec. Label any hazardous materials or waste (See resources at DEHS and ME sites)

Biological Safety Hazards 1. Risk of infection 2. Spread of biological agent to other samples or cultures 3. Contamination of work areas and equipment 4. Contamination of areas outside work area or lab

Biological Safety Hazards: Other species http: //www. ahc. umn. edu/rar/safety. html n Zoonoses n NHP – Require training to handle tissue or blood – Herpes Simplex B

Biological Safety Levels n BSL 1 – Low risk: normal uninfected samples; probability that sample contains agents pathogenic to humans is low – Includes uninfected samples from mouse, pig, dog n BSL 2 – Higher risk: samples many contain pathogenic agents that are of“moderate potential hazard” – Normal human samples, mouse with prion Tg

Biological Safety – – Non-infected animals are BSL 1 Human samples are BSL 2 – Standard Practices BSL 1 – Biosafety Level 1 (BSL-1): – Biosafety Level 1 is suitable for work involving well-characterized agents not known to consistently cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the environment. The laboratory is not necessarily separated from the general traffic patterns in the building. Work is generally conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is neither required nor generally used. Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or a related science. – The following standard and special practices, safety equipment and facilities apply to agents assigned to Biosafety Level 1:

BSL 2 in this lab Human cultures and tissues n Mouse cells with prion Tg n – This is considered BSL 2, since it has been passaged through the mouse, and is not a human prion.

Prion Tg mouse cells: Biosafety Level 2 u u “Thus, … once human prions are passaged in mice and mouse Pr. PSc is produced, these prions should be considered Biosafety Level 2 prions, even though the human prions are Biosafety Level 3 under most experimental conditions. An exception to this statement is in the case of mice expressing human or chimeric human/mouse transgenes. These transgenic mice produce human prions when infected with human prions and should be treated as Biosafety Level 2 or 3 in accord with the guidelines described above. “ From CDC

Procedures for BSL 1 n n n Wash hands after handling viable materials, after glove removal, and before leaving lab No eating, drinking in the work areas. Food stored outside work area in “food only” storage areas. Proper sharps handling procedures Minimization of aerosols and splashes Decontamination of work surfaces after use and after spill Waste (cultures, samples) – Must be decontaminated before disposal (with 10% bleach, 15 min. , or other decontamination solution, or by autoclaving) – Or, must be sealed in labeled biohazard bag/container for removal

Procedures for BSL 1 (cont. ) Biosafety hood not required at this level n Lab coats recommended n Gloves optional n Eyewear, if splashes expected n

Biological Safety for BSL 1 and BSL 2 u Sharps – Disposal in labeled needle box – Glass pipets and broken glass in labeled pipet container – Never recap needles or replace sharps in packaging – Minimize glass – Use luer lock

Biological Safety for BSL 1 vs BSL 2 u Protective clothing – Lab coat recommended, gloves optional, safety glasses (if outside hood, with risk of aerosol) u Biosafety hood – Not required u Protective clothing u Biosafety hood (and sealed rotors) – Lab coats required and are not worn outside of work areas – Gloves required – Plus face protection (if outside hood, with risk of aerosol) must be used human or other class II agents if – Aerosols or splashes – High concentration or large volume of infectious agents

Additional Procedures for BSL 2 u u Specific training for handling the pathogenic agents is required (Hepatitis B, Herpes Simplex B) Procedures that generated aerosols or splashes must be performed in biosafety hood (or similar equipment) Routing handling of human tissue and blood requires baseline serum and testing; recommended: Hepatits B immunization Any injury must be reported– Report to lab supervisor/PI – Injury form submitted – Visit to Boynton

Additional Procedures for BSL 2 (cont. ) u Decontamination of equipment and supplies – Prior to washing glassware, instruments, etc. , they must be decontaminated with 10% bleach, if not stainless steel, or with diluted decontamination solution for 15 minutes before being washed and prepared for autoclaving. – Contaminated glassware and instruments that are removed from hood should be placed into dishpan for decontamination. – Decontamination should be done immediately after completing the procedure. – To prevent any toxic effects from the decontamination solution, rinse items very well before autoclaving

Biological Safety Procedures: Human Tissue/Cells http: //www. ibc. umn. edu/principle. Jan 2005. html n Use precautions specified by Bloodborne Pathogens training – – Lab coat Gloves Neutralization of waste with 10% bleach before disposal Disposal of waste n n n Biohazard waste bag, sealed Request pick up from johns 098 Biohazard waste is autoclaved before disposal – Decontaminate hood and work area with diluted bleach, or for stainless steel, diluted decontaminant solution

Other methods for waste collection and disposal for BSL 2 n To avoid spills and aerosols, may collect waste from tubes and flasks by inserting sterile pipets into vacuum tubing set up inside hood – This must be set up with a biohazard waste trap: include bleach at 10% total flask volume, before collecting the waste

Other information n Pipet aid – Contains a filter unit inside nozzle. If medium is drawn into pipet aid, wetting filter unit, air will not pass through filter n Filter must be replaced n Filters are stored in drawer _drawer under water bath_ by hood – Occasionally, the cotton plug will be drawn into pipet aid nozzle. This can be removed with a forceps