ࡱ>   bjbj  rr/)&&DDX!|V&/"///0/3,[4L* 40044//u8u8u84F//u84u8u8B(%/>Hf95fƻ0G(/7(%%d(44u84444474444444(444444444& F:  University of Virginia Biosafety Manual & Standard Operating Procedures for Biosafety Level 2 Activities Version 2013 PI Name:  FORMTEXT       University of Virginia Charlottesville, VA IMPORTANT PHONE NUMBERS Fire and Medical Emergencies within the Medical Center .. 924-2012 Fire and Medical Emergencies..................................................911 Police.911 UVA-WorkMed (Academic staff)243-0075 UVA Employee Health (Hospital staff)..924-2013 UVA Student Health Center (students).924-5362 Hospital Emergency Room.924-2231 EHS Biosafety Office .................................................................982-4911 UVA Workers Compensation .924-8939 UVA Institutional Biosafety Committee& & & & & & & & & & ..243-0726 Principal Investigator s Emergency Contact Number......................  FORMTEXT       Evacuation Meeting Place for Laboratory Personnel:..................... FORMTEXT       Principal Investigators Certification I hereby certify that I have reviewed the contents of this manual and accompanying IBC protocol and verify that it reflects my current operating practices. I assure that all personnel under my supervision have received appropriate training for Biosafety Level 2 (BSL2) laboratory practices prior to working in this laboratory. Signature: _________________________________ Date: ______________ Signature and Acknowlegement of Risk and Laboratory Training By signing below, I certify that the PI or supervisor has explained the nature of the risks associated with the biohazardous agents that are used in the lab, the possible routes of exposure; and demonstrated the special handling, personal protective equipment (PPE), and decontamination practices that are used in the laboratory. Name (print)SignatureDate FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT        FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       Table of Contents Signatures and Acknowledgement of Risk 2 1. Purpose 4 2. Responsibilities 4 3. Experiments and Procedures Performed 5 4. Work Practices 5 A. Standard Practices for BSL2 agents 6 B. Safety Equipment 7 C. Personal Protective Equipment 7 D. Food and Drink Policy 7 E. Lab Standard Operating Procedures 8 5. Biological Waste 8 A. Biohazardous Waste (or RMW) 8 B. Other Biological Waste 8 C. Methods of Inactivation and Disposal 12 D. Human Cadavers 12 E. Hospital-like waste 12 F. Waste Laboratory Glassware 12 6. Biohazardous Spill Clean Up Procedures 12 A. Spills Outside of a Containment Device 12 B. Spills inside a Biological Safety Cabinet 13 C. Spills in a Centrifuge 14 D .Biological/Radioactive Emergencies/Spills 14 7. Proper Use of a Biological Safety Cabinet 15 8. Emergency Phone Numbers and Procedures 16 A. Fire 16 B. Injury 16 C. Exposure to Biohazardous Materials 16 D. Security Incidents 17 9. Onsite Service & Repair of Laboratory Equipment 17 10. Shipping Infectious Substances 18 Appendix A. Exposure Control Plan 20 Appendix B. Biotoxin Safety Plan 28 Appendix C. Vaccination for Microorganisms other than Hepatitis B 37 Appendix D: Policy and Procedure for Experiments Involving 48 the Use of Biological Agents in Animal Care and Use 1. Purpose This Biosafety Manual outlines procedures for conducting experiments at BSL2 containment. University laboratories approved by the UVA IBC to conduct experiments at BSL2 containment are expected to comply with the procedures in this manual. Principal Investigators or laboratory supervisors must contact the EHS Biosafety Office (982-4911) if they are uncertain how to categorize, handle, store, treat or discard any biohazardous material. The Biosafety Manual and currently approved IBC Inventory and Activity Registration (IAR) must be available and accessible to all laboratory personnel. 2. Responsibilities The Principal Investigator: Ensures that laboratory personnel demonstrate proficiency in standard and special microbiological practices before working with biohazardous agents. Ensures that all laboratory and support personnel receive appropriate training for the potential hazards associated with the work involved, the necessary precautions to prevent exposures, and post-exposure evaluation procedures. Ensures biosafety procedures are incorporated into standard operating procedures for the laboratory. Ensures personal protective equipment and necessary safety equipment is provided and used. Ensures compliance by laboratory personnel with the relevant regulations, guidelines, and policies. Reviews and updates the IBC IAR annually and the Biosafety manual as needed. Notifies the EHS Biosafety Office concerning: Any accident that results in percutaneous inoculation, mucous membrane exposure, ingestion or inhalation of biohazardous materials. Any accident involving recombinant DNA research that leads to personal injury or illness or to a breach of containment must be reported to the EHS Biosafety Office who will investigate incidents as appropriate and notify the Institutional Biosafety Committee (IBC). Any reportable incidents under the National Institutes of Health (NIH) Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules will be jointly submitted by the IBC Chair and the Biosafety Office to NIH Office of Biotechnology Activities (OBA). Any incident causing exposure of personnel or danger of environmental contamination. Minor spills not involving a breach of the biological safety cabinet (BSC) or other primary containment device that were properly cleaned and decontaminated generally do not need to be reported. Any problems pertaining to operation and implementation of biological and physical containment safety procedures or equipment or facility failure. Laboratory Personnel: Participate in appropriate training and instruction. Are encouraged to report any condition or change in health status which may increase risk or consequences of a laboratory acquired infection (e.g. pregnancy, medical conditions, medications, or treatments which compromise immunity, etc.) to UVA WorkMed (UVA employees) or Student Health (UVA students) since personal health status may impact an individuals susceptibility to infection, ability to receive immunizations, or prophylactic interventions. Additional information on worker risk for laboratory acquired infection can be found in Occupational Health and Immunoprophylaxis of Biosafety in Microbiological and Biomedical Laboratories, 5th Edition. Review and comply with biosafety procedures described in the IAR and this manual. Report all accidents, major spills, or exposure incidents immediately to their supervisor. Additional biosafety-related responsibilities for this lab (if applicable):  FORMTEXT       3. Experiments & Procedures Performed A description of the approved experiments and biological agents used in this laboratory are detailed in the attached IBC IAR. 4. Work Practices Primary hazards to personnel working in the BSL2 environment relate to accidental percutaneous or mucous membrane exposures, or ingestion of infectious materials. Special care should be taken with contaminated needles or sharp instruments. With good microbiological techniques and PPE, biohazardous agents used in the BSL2 containment environment can be used safely in activities conducted on the open bench, provided the potential for producing splashes or aerosols is low. PPE should be used as appropriate, such as splash shields, face protection, gowns, and gloves. Procedures with aerosol or splash potential that may increase the risk of personnel exposure (i.e. grinding, blending, vigorous shaking or mixing, sonic disruption, opening containers of infectious materials, inoculating animals intranasally, and harvesting infected tissues from animals or eggs) should be conducted in primary containment equipment such as a certified biological safety cabinet (BSC). In instances where a BSC is unavailable or impractical, appropriate PPE must be used to protect personnel from mucous membrane exposure (e.g. face shield with surgical mask or N95 respirator). Sinks for hand washing must be available to reduce potential environmental contamination. Standard Microbiological Practices for Biosafety Level 2 agents The PI or supervisor must enforce restricted access to the laboratory when BSL2 experiments are in progress. Access may be restricted by locking doors, posting warning signs, monitoring entry or using other suitable methods as determined by the PI. A biohazard sign (provided by the EHS) must be posted on the entrance to laboratories approved for BSL2 experiments. Persons must wash their hands after working with potentially hazardous materials, after removing gloves, and before leaving the laboratory. In instances where sinks are not immediately available, hand sanitizer dispensers may be used if approved by the IBC. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption are not permitted in the laboratory. Food must be stored outside the laboratory in cabinets or refrigerators designed and used for this purpose. Mouth pipetting is prohibited; mechanical pipetting devices must be used. Perform all procedures to minimize the creation of splashes or aerosols. Decontaminate work surfaces after completion of work and after any spill or splash of potentially infectious material with an appropriate disinfectant. Chairs and other furniture used in laboratory work must be completely covered with a non-porous material that can be easily cleaned and decontaminated with an appropriate disinfectant. Carpets and rugs in laboratories are not permitted. All cultures, stocks, contaminated wastes and other Regulated Medical Waste (RMW) are disposed of in accordance with University of Virginia Policy and Procedures (refer to Regulated Medical Waste section). A sharps management program is in place including: Needles and syringes or other sharp instruments should be restricted in the laboratory for use only when there is no alternative. Whenever practical, laboratory supervisors should adopt improved engineering and work practice controls that reduce the risk of sharps injuries. Needles, scalpels, and razor blades may only be disposed of in approved puncture-resistant sharps container. Used needles must not be bent, sheared, broken, recapped, removed from the syringe to which they are attached or otherwise manipulated by hand before disposal. Broken glassware must not be handled directly, but must be removed by mechanical means such as a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible. Cultures, tissues, specimens of body fluids, or potentially infectious wastes are placed in a container with a cover that prevents leakage during handling, processing and storage. Transport of biohazardous materials to sites within the grounds of UVA must be placed in a secondary leak proof carrier that can contain the contents if the primary container were to leak or break. Carriers must have the biohazard label affixed to the outer surface of the transport container. Contaminated equipment must be routinely decontaminated, after spills, splashes, or other potential contamination and before it is sent for repair, maintenance, or before removal from the laboratory. Spills and accidents that result in overt exposures to infectious materials, recombinant or synthetic nucleic acids or other biohazardous materials are immediately evaluated, treated, and reported to the Principal Investigator, EHS Biosafety and UVA-WorkMed. Animals not involved in the work being performed are not permitted in the lab. Additional Special Practices for this Laboratory (if applicable):  FORMTEXT       Safety Equipment All safety equipment shall be properly maintained as well as other appropriate personal protective equipment, or physical containment devices are used whenever procedures with a potential for creating infectious aerosols or splashes are conducted. These may include centrifuging, pipetting, grinding, blending, vigorous shaking or mixing, sonic disruption, opening containers of infectious materials, inoculating animals intranasally and harvesting infected tissues from animals or eggs. Biological safety cabinets, preferably Class II, will be certified annually. Centrifugation presents a physical hazard in the event of mechanical disruption. Aerosols and droplets may also be generated. High concentrations or large volumes of infectious agents may be centrifuged in the open laboratory if sealed rotor heads or centrifuge safety cups are used, and if these rotors or safety cups are opened only in a biological safety cabinet. Other Safety Equipment in this lab such as closed system sonicators, class I BSC etc.(if applicable):  FORMTEXT       Personal Protective Equipment (PPE) Laboratory coats or gowns designated for laboratory use must be worn while working with biohazardous materials. Remove lab coats before leaving for non-laboratory areas (e.g., cafeteria, library, administrative offices). All protective disposable clothing is disposed of in the laboratory. Cloth lab coats must not be taken home by personnel. If picked up and laundered by an outside vendor, UVA Procurement-approved vendors which meet criteria defined by the OSHA Bloodborne Pathogen Standard must be used. 2. Protective gloves are worn when hands may potentially contact biohazardous materials, contaminated surfaces, or equipment. Gloves are disposed of with other contaminated waste when overtly contaminated, and removed when work with biohazardous materials is completed or when the integrity of the glove is compromised. Disposable gloves are not washed or reused and they should not be worn outside the laboratory. Hands are washed following removal of gloves and before leaving the laboratory. 3. Face protection (goggles, mask, face shield or other splatter guard) is used for anticipated splashes or sprays of biohazardous materials to the face when such materials must be manipulated outside the BSC. Contaminated eye and face protection must be disposed of with other contaminated materials or decontaminated before reuse. 4. Respirators are generally not required when working in BSL2 containment. Medical clearance, fit testing and training is required to wear respiratory protection. Contact EHS for details. 5. PPE should be used in rooms containing ABSL2 animals as defined by risk assessment. Food and Drink Policy The consumption, use or storage of food and drink in laboratories or laboratory support rooms in which chemical, biological or radioactive materials are used is prohibited. Under no circumstances may food or drink be stored in refrigerators, freezers, or temperature-controlled rooms where laboratory reagents, biological specimens, animals, or other hazardous substances currently are, will be, or have been recently used or stored. Laboratory personnel are encouraged to contact the EHS Biosafety office for assistance. More information may be found on the EHS webpage. Laboratory Standard Operating Procedures Laboratory procedures are conducted in a BSC and/or with PPE as described in the attached IBC IAR. Additional procedures (if applicable):  FORMTEXT       5. Biological Waste A. Biohazardous Waste or Regulated Medical Waste (RMW): The terms biohazardous waste, infectious waste and RMW are often used interchangeably and refer to material which requires inactivation in an approved manner prior to final disposal. RMW is the term used by the Virginia Department of Environmental Quality who is the regulatory authority for biohazardous waste in Virginia. RMW includes the following materials as well as labware or other items contaminated with these materials: Microorganisms (Risk Group 2 or higher) Human blood, blood components, fluids, unfixed organs, unfixed tissues and cell lines (primary and established) Non-Human Primate materials Products of Recombinant DNA experimentation as defined by the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules. Biotoxins (with and LD 50 of less than 100 micrograms per kilogram of body weight in vertebrates)* Sharps (needles, scalpels, lancets, suture needles & items able to puncture skin)* Animal carcasses (both infected and uninfected carcasses are disposed of in the same manner) Cadaveric materials (contact EHS for information 2-4911) *Note that biologically derived toxins are in essence chemical waste, however EHS Biosafety and Hazardous Waste collaborate to provide guidance on the inactivation (e.g., autoclaving or chemical) of toxins. See Toxins Appendix for more detail. *Waste placed in sharps containers do not require inactivation prior to disposal. B. Other Biological Wastes These are biological wastes that are not known to be pathogenic to humans, are typically handled at Biosafety Level 1 (BSL1), and are not regulated by the Virginia Department of Environmental Quality (DEQ). These materials may be disposed in the regular waste stream without prior treatment; however, Principal Investigators should consider autoclaving or chemical inactivation prior to final disposal based on risk assessment and applicable grant or permit expectations. Risk Group 1 Microorganisms (typically handled at BSL1 containment and not known to cause disease in healthy humans) Tissue culture other than Human or Non-Human Primate (e.g., rodent, avian, insect, plant, etc. cells handled at BSL1) C. Methods of Inactivation and Disposal Autoclaving uses saturated steam under high pressure to decontaminate biological material (e.g. cultures, cells, contaminated pipettes, flasks etc.) and is an effective method for decontaminating biohazardous materials prior to disposal as regular trash. The PI is responsible for meeting these requirements for biohazardous waste: Maintaining a log book (documenting operator name, date, cycle time and monthly biological indicator check results) Autoclaving for a minimum of 30 minutes unless validation data shows that less time is adequate for complete decontamination Documenting proper validation using a biological indicator test (e.g., Bacillus geostearothermopholis spores). Biological indicators must be run at least once per month and testing documented. Tests should also be performed whenever new types of packaging material or trays are used, after training new autoclave users, after autoclave repair, or after any change in the waste handling process. For more details, refer to the IBC Policy on Autoclaving on the EHS Biosafety webpage. Inactivation of Liquids (Chemical) Biohazardous liquids may be decontaminated by adding bleach to a final concentration of 10% or other appropriate disinfectant to a final concentration as recommended by the manufacturer. Durable, leak proof containers must be used for liquid waste. Mix well and allow to sit for at least 15 minutes or the manufacturers recommended time. Pour decontaminated liquid into the sink and rinse with copious amounts of cold water. Liquid waste that is not compatible with certain chemical disinfectants should be decontaminated by autoclaving using slow exhaust before disposal. Contaminated Materials Containers: As an alternative to autoclaving, solid biohazardous waste may be disposed of directly into a Contaminated Materials Container (CMC). These containers meet all federal and state regulatory requirements and should be the only containers used for final disposal of biohazardous waste. CMCs are available free of charge as follows from the following locations: MR4 - Loading dock supply cabinet MR5 - Room G037C MR6 - Room G530A Jordan Hall - Room G240 Cobb Hall - Sub Basement supply cabinet Aurbach - Room 1241 Snyder - Room 171C CMCs must be lined with the provided red bags. Once a CMC is full, the top must be taped closed and properly labeled with the generators name, building, room number, phone extension and date. Depending upon the building CMCs are removed and managed by either UVA Housekeeping or the Hospitals Environmental Services Department. Laboratory personnel should avoid overloading CMCs with heavy materials and CMCs in excess of 30 pounds will not be removed by UVA Housekeeping or Hospital Environmental Services. Researchers may also take full CMCs to the appropriate waste storage area for of the building (contact EHS for location information) where it will be removed and managed by EHS. If the outside of the CMC becomes contaminated or leaks, the waste must be repackaged by placing it into a secondary container that meets all the requirements of CMCs. CMCs are picked up from central storage locations (e.g., waste rooms) by EHS and incinerated or treated offsite by a contract vendor. Sharps Careful management of needles and other sharps are of primary importance. Precautions should be taken with sharp items that include: needles, syringes with attached needles, scalpel or razor blades, scissors, lancets, guide wires, and contaminated glass Pasteur pipettes, etc. Needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal. Disposable sharps must be placed in a plastic sharps container as soon after use as possible. Sharps containers are available free of charge from EHS. Sharps containers must be located as close as possible to the area where sharps are used and should not be stored above eye level. When the sharps container is 2/3 full, close until lid clicks securely. Contact EHS to request a waste pick up by calling 2-4911 or submit a pick up request online at our website  HYPERLINK "http://www.ehs.virginia.edu/" http://www.ehs.virginia.edu/ . EHS personnel will pick up at your lab (any size sharps container will be picked up, but note that only 2 gal. sharps container replacements are provided). D. Waste Handling Chart The chart below is designed to aid you in determining how to collect, decontaminate, and/or dispose of your waste. Type of WasteBiosafety LevelProper Collection & ContainersDecontamination MethodDisposalLiquids BSL1- Non BiohazardousUse autoclavable, leak proof plastic or glass if necessary Use double containment (pan) Not Required ( HYPERLINK "http://ehs.virginia.edu/biosafety/bio.documents/Autoclaving_Guidelines.pdf" Autoclaving or Chemical Inactivation recommended) Sink (down the drain flushed w/ water) BSL2- BiohazardousUse autoclavable, leak proof plastic Use double containment - pan must be labeled with Biohazard symbol  Autoclaving Required or Chemical Inactivation Required (preferred method)  Sink (down the drain flushed w/ water)Solids BSL1- Non BiohazardousMay be deposited in regular waste/trash receptacles Place in clear autoclave bags; use double containment (pan) Not Required ( HYPERLINK "http://ehs.virginia.edu/biosafety/bio.documents/Autoclaving_Guidelines.pdf" Autoclaving recommended) Place in regular trash BSL2- BiohazardousPlace in clear autoclave bags Use double containment - pan must be labeled with Biohazard symbol   HYPERLINK "http://ehs.virginia.edu/biosafety/bio.documents/Autoclaving_Guidelines.pdf" Autoclaving Required Place in regular trashPlace in CMCNo decon: tape and label appropriatelyPlace in lab for pick up by HousekeepingSharps BSL1 or BSL2Place in sharps containers (needles, lancets, sutures, razor blades, scalpels) Other contaminated sharp-like materials that can puncture skin (e.g., broken glass, Pasteur pipettes, etc. ) must be placed in a sharps container No decon; close container when 2/3 full and complete EHS waste pick up request (online or call 2-4911) Submit request to EHS for disposal;  HYPERLINK "https://vprgsecure.web.virginia.edu/wpr/%20" https://vprgsecure.web.virginia.edu/wpr/ Animal Carcasses Non Biohazardous/ Chemical/RadPlace in opaque bag (may use vivarium provided bags) Return to vivarium freezer Disposed of by EHS ABSL2/ BiohzardousPlace in RED bag or opaque bag labeled with the Biohazard symbol  (may use vivarium provided bags with biohazard sticker) Return to vivarium freezer Disposed of by EHSOther Animal Wastes (e.g., soiled bedding, cages)Non BiohazardousPlace in opaque or paper bag Return to vivarium (designated drop off location)Disposed of by CCM ABSL2/ BiohazardousPlace in clear autoclave bag labeled (sticker) with biohazard symbol  Animal wastes (non biohazardous and ABSL2) containing blood or recognizable parts/tissues must be placed in a CMC for disposal Autoclaving Required or return to vivarium for autoclaving (designated ABSL2 drop off location) Disposed of by CCM D. Human Cadavers and/or Body Parts Human cadavers and recognizable human body parts require special handling for disposal. Please contact EHS Biosafety at 2-4911 for guidance. E."Hospital-Like" Waste (that can be disposed of as regular trash) Hospital-like wastes are containers, bags, or tubing without sharps which is not contaminated with human blood, body fluids, human derived or biohazardous materials, hazardous chemicals, or radioactivity and can be disposed of as regular trash. Examples of these materials include: urine collection bags & tubing; IV solution bags & tubing; colostomy, ileostomy, urostomy bags; plastic fluid containers, enteral feeding containers & tubing, hemovacs, urine specimen cups, oxygen tubing and cannula, nasogastric tubing, enema bags & tubing, enema bottles, and thermometer probe covers. F. Broken Glassware, Waste Laboratory Glassware (WLG) If broken glassware does not contain or is not contaminated with radioactive material (any amount), chemicals (more than a trace amount) or infectious material (any amount), then these waste materials are Waste Laboratory Glassware (WLG). WLG must be put into any ordinary cardboard box, lined with a regular trash bag. Once full, the bag top should be twisted and taped closed, the box top taped shut, and a WLG label (available from EHS) applied to the box top. These waste materials will be treated as regular trash. Glass Pasteur Pipettes: Use of glass Pasteur pipettes is discouraged. Glass pipettes should be replaced with safer alternatives (i.e., plastic) as recommended by the U.S. Department of Health and Human Services, Centers for Disease Control and Prevention and National Institutes of Health in Biosafety in Microbiological and Biomedical Laboratories, 5th Edition Fifth Edition, Feb 2007; and by The World Health Organization Biosafety Manual (WHO, 2003). Contact the Biosafety Office for more information on safer alternatives. 6. Biohazard Spill Clean-up Procedures Spills involving infectious materials or recombinant/synthetic nucleic acid molecules must be contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material. Cleaning must be done with a disinfectant effective against the agents of concern and must allow sufficient contact time for the disinfectant to work. Cleaning spills requires PPE including a lab coat or gown, and gloves. A face shield, shoe covers or a respirator may be required. All spills must be reported to the PI. Contact EHS Biosafety for any questions about spills or cleaning. Spills outside of a containment device: The spill is not inside of a Biological Safety Cabinet (BSC), Centrifuge, Refrigerator, Incubator, Freezer, Lab instrument etc. Close off spill area to traffic, and notify coworkers. If the spill may involve an aerosol, (e.g. event involving dropping material onto floor, high mechanical force, a forceful expulsion of liquid) leave the room to allow aerosols to settle. 30 minutes is the recommended wait time. Remove contaminated lab coat or clothing and wash exposed skin. Put on clean gloves and lab coat. Prepare a fresh dilution of 1:10 bleach or effective disinfectant. Prepare enough solution to saturate the entire contaminated area. Always follow manufacturers directions. If dilution of bleach is not possible, undiluted household bleach can be used, However eye protection must be worn and care taken not to splash the bleach onto skin or clothing. If contact with bleach occurs with skin, mucous membranes or eyes, flush area with copious amounts of water. Contain the spill with paper towels or other absorbent material (e.g. bench kote, blue pads, etc.) Flood the spill area with disinfectant solution and allow to remain in contact for required time: 10% Bleach for 10 minutes; or Cavicide (full strength) for 3 minutes; or Beaucoup (diluted according to manufacturers directions) for 10 minutes. Push the absorbent material at the edge of the spill into the spill's center. Add absorbent as needed. If glass is present, do not use bare hands! Use tongs (large pieces) or forceps (small pieces) followed by a dustpan to remove remaining shards. Discard the paper towels into a CMC. Discard gloves into a CMC. Washhands thoroughly. Autoclave an overtly contaminated lab coat to prior to placing into laboratory laundry bag. Report the incident to a supervisor and PI. Spills inside of a Biological Safety Cabinet (BSC) Leave BSC on. Put on gloves and lab coat and gather paper towels for cleaning. Prepare a fresh dilution of 1:10 bleach or effective disinfectant. Prepare enough solution to saturate the entire contaminated area. Always follow manufacturers directions. If dilution of bleach is not possible, undiluted household bleach can be used, however eye protection must be worn and care taken not to splash the bleach onto skin or clothing. If contact with bleach occurs with skin, mucous membranes or eyes, flush area with copious amounts of water. Flood the area with disinfectant solution and allow required contact time: 10% Bleach for >10 minutes; or, Cavicide (full strength) for >3 minutes. Spray and or wipe down the cabinet interior and any items inside the BSC with a towel dampened with disinfectant. Discard all contaminated disposable items and towels used for cleaning into a CMC or autoclave according to guidelines. Spills large enough to result in liquids flowing through the front or rear grilles require more extensive decontamination. Consult the EHS Biosafety Office immediately at 982-4911, for guidance. If a spill such as this has occurred, do not turn the BSC off. Report the incident to a supervisor and PI. Spills in a centrifuge Biohazardous spills in centrifuges can be quite difficult to disinfect. Some but not all centrifuges have closed rotors, buckets or other carriers with leak proof lids, designed to contain spills, allow efficient, safe emptying, and decontamination. However, not all centrifuges are equipped with these containment devices. A spill resulting from primary container breakage requires immediate suspension of use. PI notification and assistance from the EHS Biosafety Office. If unusual sounds from a centrifuge suggest that breakage and a spill has occurred, or, if breakage and a spill is discovered after the machine has stopped, wait at least 30 minutes after centrifuge has stopped before opening. This will allow hazardous aerosols to settle in the centrifuge. Don lab coat, gloves, and face shield prior to opening centrifuge and then open carefully to assess the situation. Use of a respirator is recommended and double gloving is advisable if glass tubes were used and broken. Attempt to determine if the spill is contained in a closed cup, bucket or tray carrier, or within a closed rotor. If the spill is contained as described in step 2, spray the exterior with disinfect and allow adequate contact time. Take the carrier to the nearest BSC approved for use with this agent. NOTE: If a BSC is not available or if the rotor cannot be removed, the centrifuge should remain closed. Post a sign indicating contaminated-do not use. Notify lab director and or PI and contact the EHS Biosafety office at 982-4911, for assistance. Obtain and place containers suitable for holding tubes, broken glass or other containers into the BSC while cleaning centrifuge components. Carefully retrieve unbroken tubes, wipe outside with disinfectant, and place them into an empty container in the BSC, out of the way. The broken glass tube(s) must be removed with a forceps or other instrument and immersed in a beaker of disinfectant solution for a time appropriate to achieve disinfection. The pieces can then be disposed of in a sharps container. After proper decontamination, carriers, rotors etc. can be washed with a mild detergent according to the manufacturers instructions. Thoroughly wipe the inside of the centrifuge chamber with disinfectant saturated towels. Allow for adequate contact time before wiping up excess liquid. Biological/Radioactive emergencies/spills The Radiation Safety Office (982-4911) must be notified and will assist in the cleanup of a biological/radioactive spill. Determine if anyone has been contaminated; remove contaminated clothing and wash contaminated skin with soap and water. Proceed with clean up as instructed by the Radiation Safety Office. The infectious agent will be neutralized first, taking care in choosing a disinfecting agent to avoid chemical incompatibility. Chlorine compounds such as bleach must NOT be used to disinfect anything containing 125I because the chlorine will cause the volatilization of radioactive iodine. Proper use of the Biological Safety Cabinet (BSC) All procedures involving the manipulation of infectious materials that may generate an aerosol should be conducted in a Biological Safety Cabinet (BSC) or other physical containment devices. BSCs must be certified annually and whenever moved to verify proper performance and air system operation. BSC are certified by an outside contactor at no cost to the Investigator. The EHS Biosafety Office provides a list of designated, NSF-certified vendors from which to choose. Contact the Biosafety Office or consult the Biosafety website for more information: Biosafety cabinets are designed to be operated 24 hours a day. It is recommended that blowers remain on at all times. If it is necessary to turn off the blower, allow sufficient time to purge airborne contaminants from the work area (Centers for Disease Control and the Public Health Agency of Canada recommend a minimum of 5 minutes before and 5 minutes after work, taking into account sufficient time for settling of aerosols). Minimize other activities in the room (e.g., rapid movement, open/closing room doors, etc.) to avoid disrupting the cabinet air barrier. Laboratory coats should be worn buttoned over street clothing. Gloves are worn to provide hand protection. Before beginning work, the investigator must adjust the stool height so that his/her face is above the front opening. Plastic-backed absorbent toweling can be placed on the work surface (but not on the front or rear grille openings). This toweling facilitates routine cleanup and reduces splatter and aerosol formation during an overt spill. Closure of the drain valve under the work surface must be done prior to beginning work so that all contaminated materials are contained within the cabinet should a large spill occur. Place necessary materials in the BSC before beginning work. This serves to minimize the number of arm-movement disruptions across the air barrier of the cabinet. All materials must be placed as far back in the cabinet as practical, toward the rear edge of the work surface and away from the front grille. The front grille must not be blocked with research notes, discarded plastic wrappers, pipeting devices, etc. Aspirator suction flasks must contain an appropriate disinfectant, and a High Efficiency Particulate Air (HEPA) in-line filter that should be replaced whenever there is evidence of filter failure or blockage. This combination will provide protection to the central building vacuum system or vacuum pump, as well as to the personnel who service this equipment. Inactivation of aspirated materials can be accomplished by placing sufficient chemical decontamination solution such as bleach, into the flask to kill the microorganisms as they are collected. Once inactivation occurs, liquid materials can be disposed of as noninfectious waste. Horizontal pipette discard trays containing an autoclave bag or an appropriate chemical disinfectant should be used within the cabinet. Upright pipette collection containers placed on the floor outside the cabinet or autoclavable biohazard collection bags taped to the outside of the cabinet should not be used. The frequent inward/outward movement needed to place objects in these containers is disruptive to the integrity of the cabinet air barrier and can compromise both personnel and product protection. All operations should be performed on the work surface at least four (4) inches from the inside edge of the front grille. Active work should flow from the clean to contaminated area across the work surface. Bulky items such as biohazard bags, discard pipette trays and suction collection flasks must be placed to one side of the interior of the cabinet. Open flames (i.e., Bunsen burners) are rarely necessary in the near microbe-free environment of a BSC. An open flame creates turbulence that disrupts the pattern of HEPA-filtered air supplied to the work surface. When deemed absolutely necessary, touch-plate microburners equipped with a pilot light to provide a flame on demand may be used. Internal cabinet air disturbance and heat buildup will be minimized. The burner must be turned off when work is completed. Microincinerators (electric) are also a possible alternative for use in the BSC. Use of ultraviolet light (UV) in the BSC is strongly discouraged. UV bulbs in the BSC must be cleaned and monitored regularly, as dust and debris inhibit effectiveness as well as gradual degradation of the UV bulb over time. Therefore, chemical surface disinfection must be the primary means of decontaminating the BSC. Upon completion of BSC experiments, a final surface decontamination of the cabinet should include a wipe-down of all interior surfaces. 8. Emergency Procedures Fire In case of fire, activate the fire alarm pull station and evacuate immediately. Judgment should be exercised in deciding whether to attempt to store or contain any hazardous materials prior to evacuation. Remove contaminated protective garments and gloves before leaving laboratory if possible. Go to designated evacuation meeting site. Injury If an injury is life threatening call 911. For less serious injuries treatment should be sought at UVAWorkMed for academic personnel, UVA Employee Health for Hospital staff or Student Health for undergraduate students during weekday, daytime hours. During night and weekend hours, treatment may be sought at the UVA Hospital Emergency Room. Any injury to a laboratory worker shall be reported as soon as possible to the Principal Investigator. Exposure to Biohazardous material An exposure is defined as biohazardous material contacting broken skin, eyes, nose, mouth, or other mucous membranes; a percutaneous injury with a contaminated sharp, or contact with an infectious agent over apparently intact skin. In the event of exposure: Wash the area with soap and water. If applicable, flush eyes, nose or mouth with water for 15 minutes. Contact UVA-WorkMed for medical assistance. Students should contact Student Health. During night and weekend hours, treatment may be sought at the UVA Hospital Emergency Room. All exposures must be reported to the immediate supervisor and PI. PIs are responsible for reporting exposure incidents to the EHS Biosafety Office who will perform a follow-up investigation of the incident and report findings to the Institutional Biosafety Committee (IBC). Some incidents may be reportable to the NIH Guideline for Research involving Recombinant DNA or Synthetic Nucleic Acid Molecules. Security incidents Security incidents such as suspicious visitors, missing chemicals, or missing biological agents must be promptly reported to the Principal Investigator. University Security or Police should be notified. PIs are responsible for reporting incidents to the Biosafety Office. 9. On-Site Servicing or Repair of Laboratory Equipment Laboratory equipment (e.g. refrigerators, freezers, incubators, centrifuges, Biological Safety Cabinets, and similar types of equipment) should be serviced only under the authorization of laboratory personnel. Laboratory personnel are responsible for decontaminating surfaces, as necessary, prior to service. Equipment can be decontaminated using the following techniques: Surface decontamination with chemical disinfectant appropriate for the agents in use Autoclaving Chemical vapor/gas decontamination: For BSC decontamination only and when accessing internal plenums or components. Scheduled and performed by an EHS approved contractor. Laboratory personnel are responsible for informing maintenance personnel regarding any potential hazards and any necessary precautions to be taken while working in the laboratory. Use of gloves, lab coat/gown or other personal protective equipment may be recommended as a precaution. Tools used for service work should be in a container that is easily disinfected (or consult with EHS as to determine the best procedure for bringing materials into the facility in the case of vivarium work). Maintenance personnel should seek recommendations of lab personnel regarding procedures to decontaminate any tools that may become contaminated while work is performed. Transfer, Removal (Decommissioning) from Laboratory or Surplus of Equipment Laboratory equipment (e.g. refrigerators, freezers, incubators, centrifuges, Biological Safety Cabinets and similar types of equipment) may be labeled with a biohazard symbol, indicating that the equipment was used in conjunction with experiments involving or used to store biohazardous agents. Accordingly, biohazard labeled equipment must be decontaminated before it can be moved outside of a UVa building, within UVA (transferred), to another institution, or sent to surplus. All surfaces and equipment (e.g., all those that are capable of tolerating chemical disinfection) must be thoroughly decontaminated by laboratory staff using an approved method. Consult with EHS Biosafety for assistance in determining decontamination procedures. Lab personnel must remove/deface biohazard symbols or stickers before equipment can be removed. Personnel must affix an EHS moving authorization label (see below) on the equipment before it is moved. UVA movers or outside contractors are not allowed to move any equipment without one of these stickers. Call EHS (982-4911) for more information or to obtain stickers. Note that yellow stickers are for UVA internal lab moves and white are for off-site or surplus removal.  10. Shipping Dangerous Goods: Infectious Substances The transportation of biohazardous substances is regulated by the Department of Transportation (DOT) and specifically air shipments are under the authority of the Federal Aviation Administration (FAA). According to the regulations, infectious substances include but are not limited to: Category A Infectious Substances Category B Biological Substances (includes human derived materials, e.g. human cell lines) Genetically Modified Organisms (GMO) and Microorganisms (GMMO) Other Biologically-derived materials (RNA, DNA, antibodies, proteins), Exempt Human or Animal Specimens, non-infectious (BSL1) microorganisms, animal cell lines (other than human and NHP which are considered infectious) are often shipped and are not regulated so long as exemption rules are followed. All shipments, whether infectious or not, that are shipped on dry ice are regulated because dry ice is considered a Dangerous Good. University personnel who ship any of the above regulated material must complete a training program. If you are uncertain as to whether or not training is needed and the material for shipment is regulated, contact the EHS Biosafety Office at 982-4911. EHS Biosafety offers an online course for Infectious Substances training and a downloadable form for Dry Ice Training HYPERLINK "../AppData/Local/Microsoft/Windows/Temporary Internet Files/AppData/Local/Microsoft/Windows/Temporary Internet Files/Content.Outlook/8OIE68TF/ehs.virginia.edu/ehs"ehs.virginia.edu/ehs. Training is valid for two years. Individuals who successfully complete training will receive a certificate confirming that they passed the final exam. Training certificates must be kept in the laboratory or must be able to be produced upon request.  FORMCHECKBOX  This Lab does not ship Category A Infectious Substances, Category B Biological Substances, Genetically-Modified Organisms, or other biological materials on dry ice.  FORMCHECKBOX  This Lab ships Category A Infectious Substances, Category B Biological Substances, Genetically-Modified Organisms, or other biological materials on dry ice. List Personnel trained, date of training and attach the training certificate: Shipping Infectious Substances and Diagnostic Specimens TrainingNameDate FORMTEXT        FORMTEXT       FORMTEXT        FORMTEXT       FORMTEXT        FORMTEXT       FORMTEXT        FORMTEXT       FORMTEXT        FORMTEXT       Appendix A Exposure Control Plan (must be completed by investigators working with human-derived materials including human cell lines) Table of Contents I. Introduction 19 II. Exposure Determination 19 Methods of Compliance 20 A. Universal Precautions 20 B. Engineering and Work Practice Controls 20-23 Communication of Hazards to Employees 23 Hepatitis B Vaccination Policy 24 Vl. Procedures for Exposure Incidents 24 VIl. Employee Training 25 Vlll. Recordkeeping Procedures 25 lX. Hepatitis B Vaccine Documentation 27 X. Sharps Safe Medical Devices in UVA Medical Storeroom 28-29 Introduction This Exposure Control Plan was prepared by the University of Virginia EHS Biosafety Office to reduce employee exposure to bloodborne pathogens in accordance with OSHA standard 29 CFR 1910.1030 and Virginia state law. This plan must be reviewed by the Principal Investigator annually and in the event of interim changes. A copy of this plan is available for review by any employee during their work shift. Principal Investigators have 15 days to provide a written copy of this plan to any employee who requests it. Exposure Determination This plan covers all employees who may reasonably be anticipated to be at risk for exposure to human blood, or other potentially infectious materials (OPIM, see list below). Employees determined to be at risk for exposure must be offered the Hepatitis B vaccine at no charge to the employee and must receive annual Bloodborne Pathogens training. Principal Investigators must determine whether an employee has the potential for exposure without considering the use of personal protective equipment (PPE). Other Potentially Infectious Materials (OPIM): Human tissue or organs (unfixed) Human tissue cultures Human blood components Blood, organs or tissue from research animals inoculated with human cell lines, human tissue Blood, organs or tissue from research animals infected with HIV, HBV, HCV or other human pathogens Semen and vaginal secretions Cerebrospinal fluid Pleural and pericardial fluid Peritoneal fluid All body fluids visibly contaminated with blood Check the job classifications in the lab with potential for exposure to human blood/OPIM:  FORMCHECKBOX  Laboratory and Research Aide  FORMCHECKBOX  Laboratory and Research Specialist  FORMCHECKBOX  Laboratory and Research Manager  FORMCHECKBOX  Post-Doctoral Fellow  FORMCHECKBOX  Professor  FORMCHECKBOX  Research Associate  FORMCHECKBOX  Other (specify):  FORMTEXT        FORMCHECKBOX  Student (specify):  FORMTEXT       Check the tasks and procedures performed by the employees listed above in which occupational exposure occurs:  FORMCHECKBOX  Handling and manipulating human blood or OPIM  FORMCHECKBOX  Other (specify):  FORMTEXT       Methods Of Compliance Principal Investigators and all staff will comply with the OSHA Bloodborne Pathogens Standard 29 CFR 1910.1030 using the following methods: Universal Precautions All human blood and OPIM are considered contaminated with bloodborne and other pathogens. Employees must avoid direct contact with human blood, and OPIM to avoid exposure to bloodborne and other human pathogens. Engineering and Work Practice Controls Engineering controls and safe work practices will be used to minimize exposure to human blood, and OPIM. Sharps and other medical devices that incorporate built-in safety features and that have been evaluated and approved by the UVA Health Sciences Center Safety Committee are available from the UVA Medical Storeroom. Safe work practices are reviewed by the Biosafety Office and the Institutional Biosafety Committee (IBC). Personal protective equipment. If the potential for exposure remains in spite of work practice and engineering controls, personal protective equipment (PPE) must be used. Employers must provide, clean, and dispose of PPE at no cost to the employee. PPE must be worn during procedures in which human blood or OPIM exposure to skin, eyes, nose or mouth is reasonably anticipated. PPE must be selected based on the type of exposure anticipated. PPE must cover all body parts and street clothes that may be exposed and must prevent soak through. Gloves, fluid-resistant gowns, face shields, masks, and other types of PPE are available from the UVA Medical Storeroom. Non-latex gloves are available for employees with latex sensitivity or allergy. PPE and personal clothing must be removed if they become contaminated. Disposable PPE that is contaminated must be discarded in a UVA Regulated Medical Waste Container (CMC). Reusable PPE such as goggles and lab coats that have become contaminated must be placed in a specified container for decontamination and reprocessing. Lab coats and gowns. Lab coats or gowns must be worn when working with human blood or OPIM when the potential for exposure exists. All PPE should be removed before leaving the laboratory for non-laboratory areas (office, library cafeteria etc.). Home laundering of lab coats and other PPE is not permitted. Contaminated laundry must be handled as little as possible and gloves must be worn. Contaminated lab coats or other PPE must first be decontaminated by soaking in a 1:10 diluted fresh chlorine bleach solution for >10 minutes or by steam sterilization (autoclaving) prior to laundering. Gloves. Gloves must be worn when hands may come in contact with human blood, or OPIM, contaminated items or surfaces. Gloves must be worn when handling animals that have been inoculated with human cell lines, human tissue, or infected with HIV, HBV, HCV, or other human pathogens. Gloves must be replaced as soon as feasible if they are torn or contaminated. Disposable (single use) gloves must not be washed or decontaminated for reuse Utility gloves may be decontaminated for reuse, but must be discarded if they are cracking, peeling or show other signs of deterioration. Glove selection should be based on an appropriate risk assessment. Protection for eyes, nose and mouth. Work must be performed in a certified Biological Safety Cabinet, or masks and eye protection (goggles or face shields) must be worn whenever splash or spray of human blood, or OPIM to the face is anticipated. Hand washing. Hands must be washed with soap and water after contact with specimens, as soon as possible after removing PPE, and whenever they become contaminated with human blood, or OPIM. Antiseptic hand cleanser may be used if soap and water are not available, but hands must be washed with soap and water as soon as feasible. Personal hygiene. Eating, drinking, smoking, applying cosmetics or lip balm, or handling contact lenses in the lab is not permitted. Food. Food and drink must not be stored in labs where human blood or OPIM are present. See policy on the EHS webpage for clarification. Pipetting. Mouth pipetting is not permitted. Minimization of aerosols. Splash, spray, spatter, or generation of droplets must be minimized during any procedure that involves human blood, or OPIM. If spattering or the generation of aerosols is reasonably anticipated, work should be performed in a certified Biological Safety Cabinet or eye protection plus a mask or face shield must be worn to prevent an exposure to the mucus membranes of the eyes, nose and mouth. Sharps handling. Careful management of needles and other sharps are of primary importance. Bending, recapping or removing needles is prohibited, except under specific infrequent circumstances. If recapping, bending or removing needles or other sharps is required by a specific procedure, and no alternative is feasible then a one handed scoop technique, mechanical device or forceps must be used. Written justification supported by reliable evidence should be included as an addendum to this Exposure Control Plan. This justification must state the basis for the Principal Investigators determination that no alternative is feasible and must describe the specific procedure that requires the recapping, bending, or breaking of needles or other sharps. Disposable sharps must be placed in a plastic sharps container as soon after use as possible. Sharps containers must be easily accessible, with the opening visible, as close as possible to the area where sharps are used and maintained upright during use. Sharps containers must be promptly closed, removed, and replaced when they are full. Close the sharp container until lid clicks securely. Contact EHS to request a waste pick up by calling 2-4911 or submit a pick up request online at the EHS website. EHS personnel will pick up sharps container at your laboratory and provide a 2 gallon sharps container replacement. Broken glass must not be picked up with bare hands. Reusable sharps, such as surgical instruments and large bore reusable needles pose the same exposure hazard as disposable sharps and must be handled in a manner similar to disposable sharps until they are reprocessed. The container used for temporary storage of contaminated reusable sharps must be puncture resistant, and labeled as Biohazard. Safe Medical Devices. Safe medical devices are used to prevent percutaneous injuries (examples may include needleless devices, shielded needle devices or plastic capillary tubes). The PI is responsible for involving employees in the selection of effective engineering and whenever practical, should adopt improved engineering and work practice controls that reduce risk of sharps injuries. Specimen Transport on Grounds. For transport to sites within the grounds of UVA, specimens of human blood and OPIM must be placed in a secondary leak proof carrier that can contain the contents if the primary container were to leak or break. Carriers must have the biohazard label affixed to the outer surface of the transport container. Servicing contaminated equipment. Before servicing or shipping, contaminated equipment must be decontaminated if possible. If it is not possible to decontaminate equipment, it must be marked with a biohazard label describing what parts remain contaminated. Central reprocessing of contaminated reusable equipment. Supplies and equipment returned to a central facility (e.g. department washroom or autoclave room) for decontamination and reprocessing must be put in a plastic bag or closeable container and marked with a biohazard label. Housekeeping. The workplace must be maintained in a clean and sanitary condition. Human blood, or OPIM spills must be cleaned up immediately with a freshly made 1:10 bleach solution or other approved disinfectant using appropriate established spill clean up procedures. Environmental Services is responsible for general cleaning in the majority of areas at the University and maintains written procedures in their office. Equipment and Working Surfaces. Contaminated work surfaces must be disinfected with 1:10 freshly made dilution of bleach or an alternative approved disinfectant. The decontamination of work surfaces must be done as soon as possible when contaminated with human blood, or OPIM, after completing procedures or at the end of the work shift if the surface may have become contaminated since the last cleaning. Temporary coverings (plastic backed paper, chux, plastic wrap, foil, etc.) over bench tops, equipment and other surfaces must be removed and replaced as soon as possible when contaminated or at the end of the work shift if the surface may have become contaminated since the last cleaning. All reusable bins, pails, cans, and similar receptacles which may become contaminated must be regularly inspected and decontaminated as soon as possible if they become contaminated. Regulated Medical Waste. Regulated Medical Waste is disposed of by UVA in accordance with the Virginia Department of Waste Management Regulations. See Biological Waste in the Biosafety Manual for guidance. Communication of Hazards to Employees Biohazard warning labels must be affixed to refrigerators, freezers, incubators and other vessels used for storing human blood, or OPIM. Containers used for transporting human specimens beyond the immediate work area must have the biohazard label affixed to the outer surface of the transport container. Biohazard labels are required on areas and equipment reasonably anticipated to be contaminated with human blood, or OPIM. Biohazard labels are not required on individual specimens that are transported inside a carrier within UVA grounds or on Regulated Medical Waste that has been decontaminated by steam sterilization. Hepatitis B Vaccination Policy Employees identified as having potential for exposure to human blood or OPIM (see the previous section Exposure Determination), must be offered the Hepatitis B vaccine at no charge to the employee. The vaccination is a series of three injections given at approximately 0, 1 and 6 months. A routine booster dose is not recommended, but will be given at no charge if the U.S. Public Health Service (PHS) recommends it in the future. The vaccine must be offered within 10 working days of initial assignment to a job category where exposure may occur. Employees who decline the Hepatitis B vaccine must sign a statement of declination located at the end of this appendix. PIs must keep a copy of this declination statement on file. Contact Work Med or Employee Health for information on receiving the vaccination series. Procedures for Exposure Incidents An exposure is defined as: blood or OPIM contact with broken skin, eyes, nose, mouth, other mucous membranes, a percutaneous injury with a contaminated sharp, or contact with blood or OPIM over a large area of apparently intact skin. In the event of exposure: Wash the area with soap and water. Flush eyes, nose or mouth with water for 15 minutes. Staff must immediately notify UVAWorkMed, UVA Employee Health, or Student Health (students only) during normal business hours. Medical personnel will advise personnel as to when they should report to the clinic for a post-exposure follow up evaluation. All laboratory personnel must report to the Hospital Emergency Room after hours. All exposures must be reported to the immediate supervisor and PI. PIs are responsible for reporting exposure incidents to the EHS Biosafety Office who will perform a follow-up investigation of the incident and report findings to the Institutional Biosafety Committee (IBC). Evaluation and Treatment of Exposures The evaluation and treatment of an exposure is confidential and will be given by or under the supervision of a licensed physician and will follow an established protocol in compliance with OSHA standard 29 CFR 1910.1030, U.S. Public Health Service, CDC guidelines, and Virginia state law. Evaluation and treatment of exposures are managed by UVAWorkMed, UVA Employee Health or Student Health Services. If the infectivity status of the source individual is unknown and blood is available, it will be tested for HIV, hepatitis B and C in accordance with state law. The exposed employee will be told what the test results are and what they mean. If the employee consents, his or her blood will be tested as soon as possible after exposure to provide baseline hepatitis B, C and HIV status. If the employee does not consent to HIV testing, the sample will be stored for 90 days and tested if the employee consents in that time period. Post-exposure prophylaxis will be offered to exposed employees when medically indicated and as recommended by the US Public Health Service. Counseling and medical evaluation will be offered for any reported illnesses the employee develops as a result of the exposure. Documentation of Circumstances Documentation of the circumstances surrounding the exposure incident is required and allows for the identification and correction of occupational hazards. Exposure incidents will be investigated by the EHS Biosafety Office in cooperation with UVAWorkMed. Employee Training All employees who may have the potential for occupational exposure to human blood, or OPIM must complete a Bloodborne Pathogens training session at the time of their initial assignment to tasks where occupational exposure may take place and annually thereafter. Annual retraining is required. Additional training must be provided whenever there are changes in tasks or procedures which affect employees' potential for exposure. Recordkeeping Procedures Medical Recordkeeping UVAWorkMed will establish a medical record for employees who have exposures. The record will be maintained for the duration of employment plus 30 years. The record is confidential and will not be disclosed to anybody within or outside the workplace without the employees written consent, except as required by law or regulation. The record will include: employee name and social security number; dates of hepatitis B vaccinations and medical records relative to the employee's ability to receive vaccination; examination results, medical testing, and followup procedures; the healthcare professional's written opinion; information provided to the healthcare professional who evaluated the employee for suitability to receive hepatitis B vaccination. Training Records The EHS web based training system will track all employees who complete Bloodborne pathogen training. Employees can access their records through the web based training system. Records will be maintained for at least 3 years. Employees, employee representatives, the Commissioner of the Virginia Department of Labor and Industry and other appropriate agencies may also request copies of these records. IX. Hepatitis B Vaccination Employees identified as having potential for exposure to human blood or other Human-derived materials must be offered the Hepatitis B vaccine at no charge to the employee. Employees who decline the Hepatitis B vaccine must sign a statement of declination. If a record of declination is not on file at UVa Work-Med, the PI should retain the following declination record. NameSignatureDateHepB vaccine Yes* or Decline** FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT      *Yes  I have already completed, I am currently undergoing, or am about to begin Hepatitis B vaccination. **Hepatitis B Vaccine Declination I understand that due to my occupational exposure to blood or other potentially infectious materials I may be at risk of acquiring hepatitis B virus (HBV) infection. I have been given the opportunity to be vaccinated with hepatitis B vaccine, at no charge to myself. However, I decline hepatitis B vaccination at this time. I understand that by declining this vaccine I continue to be at risk of acquiring hepatitis B, a serious disease. If in the future I continue to have occupational exposure to blood or other potentially infectious materials and I want to be vaccinated with hepatitis B vaccine, I can receive the vaccination series at no charge to me. Appendix B Biotoxin Safety Plan (must be completed by investigators working with toxins of biological origin) Table of Contents I. Biotoxin Safety Plan Introduction 30 II.Biotoxin Use in Research Protocols 30 A. Biotoxin Characterization 30 B. Training and Risk Assessment 31 C. Biosecurity 31 D. Safe Handling of Biotoxin 32-33 E. Spill Cleanup and Routine Decontamination 33 F. Biotoxin Inactivation and Disposal 34-35 Table 1. Biotoxin Inactivation Methods 36 G. Exposure Management 37 References 37-38 I. Introduction All Investigators working with biotoxins of biological origin must complete a laboratory specific Biotoxin Safety Plan that includes: biotoxins used, known hazards, safety equipment and protocols, personal protective equipment and training requirements, biotoxin inactivation and disposal, inventory control and security measures and emergency procedures (spills, exposure management) and medical surveillance. Guidelines for working with biotoxins of biological origin can be found in CDC/NIH Publication Biosafety in Microbiological and Biomedical Laboratories, Appendix I, Guidelines for Work with Biological Biotoxins. Biotoxin use in research protocol A. Biotoxin characterization Principal investigators, laboratory managers and personnel are encouraged to utilize the references listed below and to consult with subject matter experts before using any biotoxin. A risk assessment will help to ensure that appropriate facilities, containment equipment, policies and procedures, personnel training programs and medical surveillance protocols specific to the biotoxin and the laboratory are in place. Laboratory work with most biotoxins in biomedical research can performed safely with minimal risk to worker and community. Biotoxins are not infectious, do not replicate and are be difficult to transmit mechanically or manually from person to person. Many commonly used biotoxins have very low volatility and are relatively unstable in the environment. These characteristics further reduce the risk of biotoxins spread. The biotoxins used, total quantity (maximum amount possessed by PI at any time), description of the hazards and risks associated with these biotoxins are described in the approved Inventory Activity and Registration. 1. List any additional or unique biotoxin considerations (at discretion of PI)  FORMTEXT       B. Training and Risk Assessment Once the type and quantity of biotoxin is determined, a thorough risk assessment should be conducted to develop safe operating procedures. The main laboratory risks are accidental exposure by direct contamination of the mouth, eye or other mucous membranes; by inadvertent aerosol generation or by needle sticks and other accidents that may compromise the normal barrier of the skin. A risk assessment should evaluate laboratory facilities, equipment and procedures to be performed with biotoxins. If both biotoxins and infectious agents are used, both must be considered when containment equipment is selected and policies and procedures are written. If animals are used, animal safety practices must also be considered. Creation of pre-operational checklists is helpful and practice runs with innocuous substitutes should be conducted prior to actual biotoxin use. Each laboratory worker must be trained in the theory and practice of the biotoxin(s) to be used. Special emphasis should be placed on the nature of the practical hazards associated with laboratory operations with biotoxins. For example, the worker must be reliably and sufficiently adept at reconstitution of lyophilized biotoxin; transferring biotoxins in liquid form; required manipulation of biotoxin during experimentation; disposal of waste solutions and contaminated equipment; decontamination of work areas and after spills. C. Biosecurity An inventory control system must be in place to account for toxin use and disposition. This inventory log should list the date and time of process; name of worker; purpose of biotoxin use; amount removed and amount remaining. Primary containers should be sealed and labeled with the name and concentration of biotoxin. Secondary storage equipment (e.g. cabinets, refrigerators, freezers) should be labeled with a biohazard sticker indicating the type of biotoxin(s) and contact information for the Principal Investigator. They should be closed and locked when biotoxin is not in use to ensure restricted access. The laboratory should be closed and locked at the end of the work period. Lab doors will have hazard signs which include contact information for trained and responsible staff. Biotoxins should be used only in designated rooms approved by the Institutional Biosafety Committee (IBC) with controlled access and at predetermined lab bench areas. The room should be clearly posted with an approved UVA biohazard sign, indicating that Biotoxins are in use, authorized personnel only. Untrained personnel and visitors must be monitored and protected from biotoxin exposure. To minimize risk of exposure for untrained staff, non-related and nonessential work should be restricted in areas where and when biotoxins are in use. Usage of biotoxins at a quantity or in procedures with increased risk (i.e. handling or reconstituting lyophilized toxin) should be conducted only when another trained individual is available to observe processes. Any unexpected, undocumented loss of biotoxin should be reported immediately to the PI, the EHS Biosafety office, and UVA police. Describe the inventory control and security procedures put into place for biotoxin use:  FORMTEXT       D. Safe handling of biotoxin Engineering controls should be used during any handling of biotoxin. Routine operations with dilute biotoxin solutions are conducted under BSL2 conditions with a certified biological safety cabinet (BSC) or comparable engineering controls such as a chemical fume hood (CFH). A certified BSC or CFH will suffice for routine operations with most biotoxins. Work with low molecular weight biotoxins or solutions which combine biotoxins with volatile chemicals or radionuclides may require the use of a charcoal based hood filter in addition to HEPA filtration. All work with biotoxins should be conducted within the operationally effective zone of the chemical fume hood or BSC with verification of inward airflow prior to work. Vacuum lines must be protected with HEPA filter to prevent entry of toxins into the line. Selected operations may require modified BSL-3 practices and procedures. This determination is made in consultation with biosafety personnel, considering all variables: biotoxin in use, physical state (solution or dry form), and the total amount of biotoxin used relative to human lethal dose, volume of material manipulated, methodology, human and equipment performance limitations. Personnel protective equipment When using an open front fume hood or BSC, PPE must protect the hands and arms. When working with biotoxins that poses dermal hazards, gloves which are impervious to the biotoxin and diluents should be carefully considered. When conducting liquid transfers and operations which pose a splash or droplet hazard; safety glasses, disposable face mask or face shield must be worn. Static free gloves should be worn when handling dry forms of toxin. If it is necessary to reconstitute lyophilized biotoxin, the task must be performed in a BSC or CFH. If a rubber septum is not in place to prevent accidental release of biotoxin powder, the worker must wear respiratory protection. Medical evaluation, fit testing and training are required prior to wearing respiratory protection. Contact EHS for details at 982-4911. Biotoxin preparation for laboratory use Working with Solutions: Solutions should be transported in leak/spill proof secondary containers to and from the hood/BSC. Before removing from the BSC or CFH, closed primary containers must be decontaminated and then placed into clean secondary container. Working with dry forms: Lyophilized biotoxin should be reconstituted in a manner so as to reduce the chance of the release of dry electro statically charged particles or liquefied splash or droplet production within the BSC or CFH. It is recommended that the new, unopened vial be physically stabilized (e.g. inserted into a block of Styrofoam or foam rubber) during the entire process. Double, static-free gloves should be worn so that one pair may be removed if a visible dusting or splash occurs. The cap or lid on the vial should be covered with damp gauze or tissue to capture any material which may be released. If a rubber septum is not in place to prevent accidental release of biotoxin powder, the worker must wear respiratory protection during the process. Medical evaluation, fit testing and training are required prior to wearing respiratory protection. Contact EHS for details at 982-4911. Mechanical forces: Operations that expose biotoxin solutions to vacuum or pressure, e.g. membrane filter sterilization or pressurized tubes should be conducted in a BSC or chemical fume hood. Centrifugation of cultures or materials potentially containing biotoxins should only be performed using sealed, thick-walled tubes in safety centrifuge cups or sealed rotors. A description of the precautions taken to prevent exposures (e.g., PPE, BSC, CFH) is included in the IAR for this laboratory. Prevention of mechanical injuries and use of sharps Efforts should be made to reduce or minimize the use of any sharps during manipulation of biotoxins. Only workers trained and experienced in handling animals should be permitted to conduct operations involving biotoxin injection. Glassware should be replaced with plastic wherever possible. Thin walled glass equipment (e.g. Pasteur pipettes), should be completely avoided. Glass chromatography columns under pressure must be enclosed within a plastic water jacket or other secondary container Spill cleanup and routine decontamination Guidelines for decontamination of selected biotoxin preparations or materials contaminated with biotoxins are summarized in Table 1 Biotoxin Inactivation Methods and review of extensive references included in the BMBL, 5th ed. is advised. Inactivation or decontamination procedures should not be assumed to be 100% effective without validation using specific biotoxin bioassays. Depending upon the biotoxin, contaminated materials and biotoxin waste solutions can be inactivated by incineration or autoclaving or by soaking in suitable decontamination solutions e.g. NaOCl and or NaOH. Spill cleanup: Appropriate biotoxin inactivation solutions should always be readily available in the event of a spill. If a spill occurs, personnel should warn others to stay away from the area, don PPE, and fully assess the extent of the spill. In the event of a large spill, consultation with OEHS biosafety personnel is advised. Avoid splashes or aerosol generation by covering spill with disposable, absorbent material (e.g. paper towels). Apply appropriate decontamination solution starting at the perimeter of the spill and working to center. Allow sufficient time for biotoxin to be inactivated. Contaminated or potentially contaminated protective clothing should be decontaminated using suitable chemical methods or autoclaving. If this is impractical, they should be disposed of as hazardous materials. Significant spills should be reported to the Biosafety Office. 982-4911 Describe decontaminating (work surfaces, equipment, devices) and spill cleanup procedure  FORMTEXT       F. Biotoxin inactivation and disposal Material safety data sheets (MSDS) which accompany all commercial biotoxin preparations may include information on biotoxin deactivation or spill cleanup. Suggested procedures follow for the methods referenced in Table 1 Biotoxin Inactivation Methods. Steam Heat: Biotoxin stability varies considerably depending upon its: temperature, pH, ionic strength, availability of co-factors and other characteristics of its surrounding matrix. Some biotoxins are inactivated by autoclaving for one hour at 121C, while others are inactivated by exposure to sodium hypochlorite or a mixture of sodium hypochlorite + sodium hydroxide. Generally, the higher molecular weight proteinacious bacterial biotoxins are inactivated by steam sterilization. Steam sterilization should not be used for destruction of any low molecular weight biotoxins (e.g. mycotoxins, marine and reptile venoms). Steam Sterilization (Autoclaving) of biotoxins Do the work in a BSC or CFH. Wear a long sleeved lab coat or gown, gloves and eye protection. Loosen the cap of the primary biotoxin container to allow steam penetration. Place the primary container into a secondary disposable container (e.g., small sharps container). Place the sharps container in a loosely closed autoclave bag. Place the bag in an autoclavable pan. Autoclave at 121 C for 1 hour on liquid cycle (slow exhaust). Discard the biohazard bag with its contents in a contaminated materials container (CMC) Dry Heat: Literature values for dry heat inactivation can be misleading due to: variations in experimental conditions; non-linear function of heat inactivation methods; protein re-folding and partial reverse of heat inactivation; disparity in denaturing a biotoxin in aqueous solutions versus a dry powered preparation. Dry Heat Inactivation Loosen the cap of the primary biotoxin container to prevent breakage. Place the primary container into a secondary disposable container (e.g., small sharps container). Place the sharps container in a loosely closed autoclave bag. Place the bag in an autoclavable pan. Heat to recommended temperature for 1 hour. Discard the biohazard bag with its contents in a CMC. Chemical Destruction of biotoxins (e.g., NaOH, and or NaOCl) When using sodium hypochlorite or a mixture of sodium hypochlorite plus sodium hydroxide to destroy biotoxins, work in a BSC or Chemical Fume hood, and wear a long sleeved lab coat or gown, gloves and eye protection. Cover the work surface with plastic backed absorbent paper (bench diaper) Put the biotoxin into solution, if not already in liquid form. Place the biotoxin container in a secondary stabilizing container Add an equal volume of the sodium hypochlorite (or sodium hypochlorite + sodium hydroxide) to the primary container of biotoxin solution. Do not replace the cap on primary container. Allow a minimum 60 minutes exposure time (Refer to BMBL 5th ed. for additional exposure time recommendations). Secure the cap on the primary container. Double bag the material in zip-lock plastic bags and label it Inactivated/denatured (BIOTOXIN NAME). Complete a waste slip listing the biotoxin as inactivated and contact OEHS (982-4911) for disposal as hazardous waste. Note: A reduction in paperwork and pickups can be achieved by collecting decontaminated biotoxin in one container, for one pick up, as long as it does not exceed the 90 day time requirement for pickup of hazardous materials. Describe procedures for biotoxin inactivation  FORMTEXT       Table 1. Biotoxin Inactivation Methods ToxinAutoclave e"121p C for 1 hr Dry Heat (10 min.)NAOCL (30 min) NAOH (30 min)NAOCL + NAOH (30 min)Abrin <100mg/PI*YesNoNoAnthrax Lethal Toxin (PA & LE)10%Bungarotoxin Methods vary depending on structure, see MSDS provided by vendorBotulinum neurotoxins<0.5mg/PI*Yes  > 100 C >0.1 %  >0.25 NNDBrevetoxin (PbTx-2) No > 815 C e"2.5%ND0.25% + 0.25NClostridium perfringens epsilon toxin <100mg/PI*YesYesYesConotoxin Methods vary depending on structure, see MSDS provided by vendorDiacetoxyscirpenol (DAS), T-2 <1000mg/PI*No > 815 C e"2.5%0.25% + 0.25NMicrocystines No >260 Ce"0.5%ND0.25% + 0.25NPalytoxin No >260 Ce"0.1%ND0.25% + 0.25NPertussis toxin Acids or bases can be used, see MSDS provided by vendorRicin <100mg/PI*Yes > 100 C>1.0%ND>0.1% + 0.25NSaxitoxin <100mg/PI*No >260 Ce"0.1%ND0.25% + 0.25NShiga toxin & Shiga-like ribosome inactivating proteins <100mg/PI* Yes Yes YesStaphylococcal enterotoxins<5mg/PI*Yes > 100 C; refolds > 0.5%>0.25 NNDTetrodotoxin <100mg/PI*No >260 Ce"0.5%ND0.25% + 0.25NDiphtheria Toxin1 YesYes YesYesTetanus Toxin2,3 Yesn/an/a*Some biological toxins are classified by the Federal Government as Select Agents due to their potential to pose a severe public health threat. The possession, use and transfer are highly regulated. In the quantities listed, they are exempt from regulation. Call the Biosafety Office for guidance and consult  HYPERLINK "http://www.cdc.gov/od/sap" http://www.cdc.gov/od/sap. 1. A solution of 0.25N 1 N NaOH may be used for surface decontamination 2. 2.5% NaOCL may be used for surface decontamination 3. Although some technical references provide recommendations for chemical inactivation and surface decontamination, there is no consensus regarding the most effective type and concentration. Thus, inactivation by autoclaving is recommended. (List Biological Laboratories) G. Exposure management All accidents, incidents, spills, exposures, or injuries, must be immediately reported to the Principal Investigator. Thorough documentation of the event is advised. Workers requiring medical attention should contact WorkMed (243-0075). Students requiring medical attention should contact Student Health (924-5362). After hours, workers needing medical attention are to go to the UVA emergency department. Medical personnel will advise if a post-exposure follow up is required. The Principal Investigator is responsible for reporting any accidents, incidents, spills, exposures, or injuries to the Biosafety office. The Biosafety Office will perform a follow up investigation and report its findings to the IBC. References: American Industrial Hygiene Association - Biosafety Reference Manual. Heinsohn P.A.; Jacobs R.R.; Concoby B.A. (eds.). American Industrial Hygiene Association, Fairfax. 1995. Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, U.S. Department of Health and Human Services, Centers for Disease Control and Prevention and National Institutes of Health, Fifth Edition, Feb 2007.  HYPERLINK "http://www.cdc.gov/biosafety/publications/bmbl5/index.htm" www.cdc.gov/biosafety/publications/bmbl5/index.htm Biological Defense Safety Program, Pamphlet 385-69, Final Rule. Dept. of the Army, 1993.  HYPERLINK "http://www.armystudyguide.com/content/publications/da_pams/da-pam-38569.shtml" http://www.armystudyguide.com/content/publications/da_pams/da-pam-38569.shtml Biological Safety Principles and Practices, 4th Edition, Edited by Diane O. Fleming and Debra L. Hunt ASM Press 2006. Cordova, M.D., Raul Flores. "The Antitoxic Power of Hypochlorous Acid." The British Medical Journal (1915): 504-5. CRC handbook of Toxicology. Derelanko M.J., Hollinger M.A. (eds.). CRC Press, Boca Raton 1995. Department of the Army, DOD. 32 CFR Parts 626, 627 - Biological Defense Safety Program. DHHS/ CDC Select Agent Program:  HYPERLINK "http://www.cdc.gov/od/sap/" http://www.cdc.gov/od/sap/ Ellenhorn's Medical Toxicology: Diagnosis and Treatment of Human Poisoning. Ellenhorn M.J., Schonwald S., Ordog G., Wasserberger J.Williams and Wilkins, Baltimore 1997. Factsheets on Chemical and Biological Warfare,  HYPERLINK "http://www.cbwinfo.com/Biological/BWList.shtml#Toxins" http://www.cbwinfo.com/Biological/BWList.shtml#Toxins Field Management of Chemical Casualties Handbook, 2nd Edition, 2000. Chemical Casualty Care Division, USAMRICD, Aberdeen Proving Ground, Maryland.  HYPERLINK "https://www.rke.vaems.org/wvems/Libraryfiles/Dis/E_04.pdf" https://www.rke.vaems.org/wvems/Libraryfiles/Dis/E_04.pdf Franz, David R. Franz, DVM, Ph.D, 1997. Defense Against Toxin Weapons, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland. Johnson Barbara, Mastnjak Ray, Resnick I. Gary, Safety and Health Considerations for Conducting Work with Biological Toxins, Applied Biosafety 6(3) pp117-135, American Biological Safety Association, 2001. List Biological Laboratories MSDS for tetanus toxin product #190 and diphtheria toxin product #150.  HYPERLINK "http://www.listlabs.com" www.listlabs.com National Research Council. Prudent Practices in the Laboratory: Handling and Disposal of Chemicals. National Academy Press, Washington D.C. 1995. Potential Biological Agent Operational Data Charts, Adapted from FM8-9: NATO Handbook the Medical Aspects of NBC Defensive Operations AMedP-6(B).  HYPERLINK "http://www.cbwinfo.com/Biological/FM9Table.html" http://www.cbwinfo.com/Biological/FM9Table.html United States Department of Labor, Occupational Safety and Health Administration. 29 CFR Part 1910 Occupational Safety and Health Standards. USAMRIIDs Medical Management of Biological Casualties Handbook (The Blue Book) 6th Ed, April 2005. U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland.  HYPERLINK "http://www.usamriid.army.mil/education/instruct.cfm" www.usamriid.army.mil/education/instruct.cfm Wannemacher R.W. 1989. Procedures for Inactivation and Safety Containment of Toxins. Proc. Symposium on Agents of Biological Origin, U.S. Army Research, Dev. and Engineering Center, Aberdeen proving Ground, MD. pp. 115-122. Appendix C Vaccination for Microorganisms other then Hepatitis B (must be completed by investigators working with Vaccinia, Rubella, N. meningitis or microorganisms (other then Hepatitis B) or toxins for which a vaccine is available). The Hepatitis B vaccination page can be found in Appendix A. Employees identified as having potential exposure to (specify microorganism or biotoxin)  FORMTEXT      must be offered the vaccine at no charge to the employee. Employees who decline the vaccine must sign a statement of declination. Principal Investigators must keep a copy of the declination statement on file. NameSignatureDatevaccine (specify) Yes* or Decline** FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       FORMTEXT       *Yes  I have already completed, I am currently undergoing, or am about to begin vaccination. ** Vaccine Declination I understand that due to my occupational exposure to (specify microorganism):  FORMTEXT      . I may be at risk of acquiring infection. I have been given the opportunity to be vaccinated at no charge to myself. However, I decline vaccination at this time. I understand that by declining this vaccine I continue to be at risk of acquiring infection, if in the future I continue to have occupational exposure and I want to be vaccinated, I can receive the vaccination series at no charge to me. Appendix D Policy and Procedure for Experiments Involving the Use of Biological Agents in Animal Care and Use Experiments that involve the administration of biological agents require special precautions to protect human health and the environment. The precautions described in this policy represent the minimum requirements for animal experiments involving biological agents and follow recommendations set forth in the NIH Guide for the Care and Use of Laboratory Animals (National Academy Press, 1996), Occupational Health and Safety in the Care and Use of Research Animals (National Research Council, 1997), and the CDC-NIH Guideline Biosafety in the Microbiological and Biomedical Laboratories. In addition to adhering to the requirements presented in this document, it is essential that animal handlers and vivarium supervisors read and understand all protocol-specific requirements before beginning an experiment. Policy Statement: In accordance with University  HYPERLINK "https://policy.itc.virginia.edu/policy/policydisplay?id=SEC-012" Biosafety Policy SEC-012, The University of Virginia will comply with all applicable federal, state and local regulations which apply to the possession, manipulation and disposal of biological agents. Additionally, consensus biosafety guidelines such as those from the National Institutes of Health, Centers for Disease Control, and other organizations will be applied as appropriate to protect personnel and the environment from potentially adverse exposures to biological agents. Consistent with the policy statement, the following Animal Biosafety Level 2 (ABSL2) procedures and precautions are provided to aid personnel in understanding the expectations of the Animal Care and Use Committee. Approval Experimentation involving the following types of biological agents will require  HYPERLINK "https://vprgsecure.web.virginia.edu/bio/home.cfm" Institutional Biosafety Committee (IBC) approval prior to ACUC approval: Risk Group 2 (and above) microorganisms. These are agents associated with human disease that is rarely serious and for which preventive or therapeutic interventions are often available. Examples include: Entamoeba sp., Salmonella sp., Adenovirus, Herpesvirus, Helicobacter pylori. Viral vectors (e.g. Herpesvirus, Adenovirus, Vaccinia, Sindbis, Lentivirus, etc.). Materials of human origin including blood, fluids, tissues, primary and established human cell lines. Recombinant DNA activities required to be conducted at BSL 2 containment as described in the  HYPERLINK "http://oba.od.nih.gov/rdna/nih_guidelines_oba.html" NIH Guidelines for Research Involving Recombinant DNA Molecules. Biotoxins with an LD50 of less than 100 micrograms per kilogram of body weight in vertebrates (e.g. Tetrodotoxin, Ricin, SEB, Diptheria Toxin, Botulinum toxin) Generation of rodents in which the animal's genome has been altered by stable introduction of recombinant DNA, or DNA derived therefrom, into the germ-line (transgenic animals). Note that these experiments require IBC notice simultaneous with initiation. Refer to the IBC webpage or contact the  HYPERLINK "http://keats.admin.virginia.edu/" Office of Environmental Health and Safety (OEHS) at 982-4911 for assistance in determining appropriate biosafety levels. Special Approval Considerations: Animal experiments involving  HYPERLINK "http://www.selectagents.gov/agentToxinList.htm" Select Agents and Toxins as defined by 42 CFR 73 must be approved by the Responsible Official (RO) prior to ACUC approval. Contact OEHS for additional communication with the RO. Animal experiments conducted at Animal Biosafety Level 3 (ABSL3) are subject to biosafety and biosecurity measures that extend beyond the ABSL2 precautions described in this document. Personnel must be approved to work in the BSL-3 facility by its director and must comply with ABSL3 work practices and procedures as defined by the BSL-3 director and ACUC policies. Medical Considerations All animal handlers are required to participate in the Occupational Health and Safety Program for animal research at University of Virginia. Personnel must comply with medical clearance and surveillance program requirements as determined by the IBC and ACUC. Training Personnel must have specific training in animal facility procedures, the handling of infected animals and the manipulation of pathogenic agents. In addition to training requirements defined by the ACUC and IBC, Principal Investigators and laboratory supervisors are responsible for ensuring that personnel under their supervision are adequately trained to perform the experiments in a safe manner. Hazard Notification and Communication Investigators are required to notify the appropriate vivarium supervisor prior to introducing biological agents into animal facilities. Supervisors will make arrangements to post biohazard warning signs at the entry of vivaria rooms where affected animals will be used and housed. The sign must include the animal biosafety level, general occupational health requirements (e.g. vaccination), personal protective equipment requirements, the supervisors name (or other responsible personnel), during work and after hours contact telephone numbers, and required procedures for entering and existing the animal areas. In animal housing areas and/or procedure rooms that are under the direct supervision of the investigator, the investigator is responsible for posting biohazard signage at the immediate entry to the room. Signs may be obtained from OEHS. When experiments involve rodents, investigators are responsible for ensuring that animal cage cards properly identify the biological agent and are marked with the universal biohazard symbol for the duration in which biological agents may be present in the cage. Investigators should consult with the vivarium supervisor to determine suitable hazard communication for experiments involving the administration of biological agents to non-rodent species. Soiled caging must be appropriately contained and identified as biohazardous if returned to the vivarium for processing in a cage wash area. Restricted Access Access to the animal room must be limited to those authorized persons required for program or support purposes. It is the responsibility of the vivarium supervisor (or laboratory director) to restrict access and ensure that all personnel, including vivarium personnel, service workers, and visitors are advised of the potential hazards (natural or research pathogens, allergens, etc.) and are instructed on the appropriate safeguards. Personal Protective Equipment Personal protective equipment (PPE) including, but not limited to, disposable gowns (or lab coats), and protective gloves must be worn when handling infected animals. Additional PPE for particular rooms, biological agents, or animal species may be required as defined by the ACUC, IBC, OEHS, or veterinary staff. For example, face/eye protection and surgical masks or respiratory protection must be worn by all personnel entering animal rooms that house nonhuman primates. Personnel using respirators must be enrolled in the University  HYPERLINK "http://ehs.virginia.edu/os/resp.html" Respiratory Protection Program, which includes medical qualification; respirator fit testing and respirator use training. 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((044 lap(X.\&'2Ɔ) xxxxg & Fdd[$\$gd`-m$ dd[$\$gdSgd9$a$gdgd5 Y`0p@ P !$`'0*-/2p5@8;=@CPF Id,gd2 Y`0p@ P !$`'0*-/2p5@8;=@CPF Igd 24Z\ %&'12huhVDh2#hwjShu5CJOJQJ^JaJ#hwjShS5CJOJQJ^JaJ#hwjShf5CJOJQJ^JaJhwjShOJQJ^Jhc/OJQJ^J2jhwjSh5>*OJQJU^JmHnHu-jmbhwjSh5>*OJQJU^JhwjSh5>*OJQJ^J'jhwjSh5>*OJQJU^JhwjShOJQJ^JhwjSh5OJQJ^JaJhwjShOJQJ^JaJhiDEF^_ Z[[ɸwgYEw'jchwjShSOJQJU^JhwjShS5OJQJ^JhwjShS5>*OJQJ^JhwjShS0JOJQJ^J'jbhwjShSOJQJU^J!jhwjShSOJQJU^JhwjShSOJQJ^J!hwjShS5;OJQJ\^J#hwjShS5CJOJQJ^JaJ#hwjShu5CJOJQJ^JaJ#hwjSh95CJOJQJ^JaJ V@A5̒ѓғۓjk<=Hdd[$\$^Hgdfm$ & Fdd[$\$gd`-m$ dd[$\$gdS & Fdd[$\$gd`- dd[$\$gdfm$[\]ijÏďRS̒ѓғۓjkǕϕ nu<ھھھھھyyk\\\hwjShS0J(OJQJ^JhwjShS5OJQJ^JhwjShfOJQJ^JhwjShS5>*OJQJ^J'jfhwjShSOJQJU^J'jehwjShSOJQJU^JhwjShSOJQJ^JhwjShS0JOJQJ^J!jhwjShSOJQJU^J'jdhwjShSOJQJU^J"=>P"':7=q dd[$\$gdS dd[$\$gdfm$ & Fdd[$\$gd`-m$ dd[$\$gdfm$<>OP!"=>pqrDEF\]ƹƨƔxvƨbShwjShS0J(OJQJ^J'jKhhwjShSOJQJU^JUhwjSh=OJQJ^JhwjShS0JOJQJ^J'jxghwjShSOJQJU^J!jhwjShSOJQJU^JhwjShfOJQJ^JhwjShSOJQJ^JhwjShS5OJQJ^JhwjShS5>*OJQJ^Jhfhf5OJQJ^J re information. Protective clothing and equipment must not be worn beyond the boundary of animal work areas or taken home for any reason. Work Practices Eating, drinking, handling contact lenses, applying cosmetics and storing food for human use is not permitted in animal or procedure rooms. Consideration should be given to the use of restraint practices and devices that reduce risk of exposure during ABSL2 animal manipulations (e.g. physical restraint devices, chemical restraint medications, etc.).Administration of biological agents requiring ABSL3 containment will be conducted while animals are anesthetized or physically immobilized in a manner which minimizes risk of biological exposure to personnel, or release of biological agents into the environment, unless scientific justification exists to do otherwise. All procedures must be carefully performed to minimize the creation of aerosols or splatters of infectious materials and waste. Properly maintained biological safety cabinets, personal protective equipment (e.g. gloves, lab coats, face shields, respirators, etc.) and/or other physical containment devices or equipment are used whenever conducting procedures with potential for creating aerosols or splashes. These include necropsy of infected animals, harvesting of tissues or fluids from infected animals or eggs, or intranasal inoculation of animals. In certain cases where physical containment devices are unavailable, combinations of personal protective equipment (e.g. respirators and face shields) and work practices may provide satisfactory protection. Such instances must be defined in the ACUC protocol application and approved by the ACUC prior to initiating experiments. All personnel must wash hands after handling animals or animal tissues and prior to leaving animal care and use areas, even if gloves are used. If a sink is unavailable for hand washing, an alcohol-based hand sanitizer (or other sanitizer effective against the agent) may be used but hands should be washed with soap and running water as soon as feasible. Equipment and work surfaces must be decontaminated with an effective disinfectant (e.g. Spor-Klenz, Cavicide, 10% bleach) after work with the infectious agent, and after any spill or splash of viable materials. A list of  HYPERLINK "https://vprgsecure.web.virginia.edu/bio/disinfectant_summary.html" suitable disinfectants and their properties is available from OEHS. Spills and accidents that result in overt exposure to infectious materials must be immediately reported to the vivaria and/or individual supervisor. Supervisors are responsible for reporting incidents to the Institutional Biosafety Officer at 982-4911. All infectious materials must be collected, labeled, transported, and processed in a manner that contains and prevents transmission of the agent(s). The outer surfaces of potentially contaminated containers must be disinfected prior to moving the material. Use of Sharps Use of needles and syringes or other sharp instruments are permitted in the vivaria only when there is no alternative, such as for parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles. Syringes that re-sheathe the needle, needle-less systems, and other safe devices should be used when possible. Contact OEHS for guidance regarding safe needle devices. Plastic-ware should be substituted for glassware whenever possible to minimize the risk of breakage. Broken glassware must not be handled directly; it should be removed using a brush and dustpan, tongs, or forceps and should be decontaminated prior to disposal. Disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated before disposal. Used needles must be carefully disposed of directly into an appropriately designated puncture-resistant sharps container located as close to the work site as possible. Animal Transport Animals must be transported in a manner that contains and prevents transmission of infectious agents during transport. The outer surfaces of potentially contaminated containers must be disinfected prior to transport. Refer to the Center for Comparative Medicine for guidance related to vehicular transport or off grounds transport of animals. Waste Disposal Animal carcasses and tissues generated from ABSL2 experiments must be placed into a red plastic bag or opaque bag marked with the biohazard symbol and returned to the vivaria freezer or cold room designated for carcass disposal. Bags for disposal may be obtained from vivaria supervisors Contaminated bedding and caging from ABSL2 experiments must be autoclaved prior to cage cleaning. Investigators should autoclave soiled ABSL2 cages before returning them to the vivarium. In instances where cage autoclaving cannot be performed by the investigator, cages must be placed into an autoclave bag clearly labeled with the universal biohazard symbol, and returned to the vivarium for processing. Intra-vivarium transport of soiled ABSL2 cages must be performed in a manner that the cage and/or autoclave bag containing the cage is clearly identified with the universal biohazard symbol and the potential for spillage is minimized (i.e. do not attempt to hand carry an excessive number of cages). ABSL3 carcasses, tissues, caging systems, sharps and other refuse must be steam sterilized (autoclaved) before removal from ABSL3 areas. Alternative means of decontamination require prior approval by the IBC and/or ACUC. All infectious wastes from the animal room (including animal tissues, carcasses, contaminated bedding, unused feed, sharps, and other refuse) must be transported in a leak-proof, covered container in a manner that contains and prevents transmission of the agents. The outer surfaces of the potentially contaminated containers must be disinfected prior to moving the material. Sharp items (e.g. syringes, needles, scalpels, razors, etc.) must be disposed of in appropriately designated puncture-resistant sharps containers. Refer to the EHS,  HYPERLINK "http://ehs.virginia.edu/tree/home.html" Waste Management Decision Tree for more information regarding waste disposal. Reporting Incidents, Injuries and Illnesses All exposures, injuries, illnesses, animal bites and scratches, needle sticks, allergic reactions, accidents, etc. must be immediately reported to your supervisor.  HYPERLINK "http://www.healthsystem.virginia.edu/internet/occupational-health/home.cfm" UVA-WorkMed (243-0075) or or  HYPERLINK "http://www.virginia.edu/studenthealth/" Student Health Services (982-3915) should be notified to determine if medical attention is warranted. After hours incidents should be referred to the Hospital Emergency Room. Incidents involving macaques (monkeys of the genus Macaca), which have the potential to carry B-virus (Herpesvirus simiae), must also follow the emergency procedures detailed in Comparative Medicines SOP#702 and CDC, USPHS recommendations contained in the Monkey Bite & Scratch Kit maintained adjacent to these monkeys at their vivarium. Individuals who observe conditions or work practices that pose a potential risk should notify their supervisor. Supervisors should communicate these observations to the Office of Environmental Health and Safety for further investigation and potential mitigation. Additional Information For more information regarding precautions for handling biological agents, contact the Institutional Biosafety Officer at 982-4911.     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