ð EXPOSURE  CONTROL  PROGRAM    BIOLOGICAL  SAFETY  LEVEL  2  PRACTICES  AND  PROCEDURES  

DARTMOUTH  COLLEGE  ENVIRONMENTAL  HEALTH  AND  SAFETY  

 JULY  2011,  VERSION  1  

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Table  of  Contents  

INTRODUCTION   4  

SCOPE   4  RESPONSIBILITIES   5  

DETERMINING  RISKS  OF  EXPOSURE   5  

PLANNING  FOR  RISK  ASSESSMENT  OF  EXPOSURE   5  OTHER  RISK  DETERMINATION   6  

METHODS  OF  EXPOSURE  CONTROL   7  

GENERAL   7  POSTING  HAZARD  REQUIREMENTS   7  ENGINEERING  HAZARD  CONTROLS   8  WORK  PRACTICE  CONTROLS   10  PERSONAL  PROTECTIVE  EQUIPMENT   10  

HOUSEKEEPING   12  

GENERAL   12  DECONTAMINATION  AND  SPILL  CLEAN-­‐‑UP   12  

SPILL  CLEAN-­‐‑UP  PROCEDURE   12  

BIOLOGICAL  WASTE  DISPOSAL   13  

EMERGENCY  STEPS  TO  TAKE  IN  THE  EVENT  OF  BIOLOGICAL  EXPOSURE   13  

MEDICAL  SURVEILLANCE  AND  POST-­‐‑EXPOSURE  EVALUATION   14  

THE  DARTMOUTH  COLLEGE  EXPOSURE  CONSULTATION  PROGRAM   14  ANIMAL  EXPOSURE  SURVEILLANCE  PROGRAM   14  POST-­‐‑EXPOSURE  EVALUATION   15  HEPATITIS  B  VIRUS  AND  HBV  VACCINATION   15  

EMPLOYEE  NOTIFICATION   15  

RDNA  REGISTRATION   16  

ENHANCED  BIOLOGICAL  SAFETY  LEVEL  2  (BSL2)   16  

TRANSPORTATION  OF  BIOLOGICAL  MATERIALS   17  

GENERAL   17  SHIPPING  CATEGORIES   17  

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EQUIPMENT  REPAIR  AND  TRANSFER   18  

TRAINING  AND  EDUCATION   18  

INSTITUTIONAL  BIOSAFETY  COMMITTEE   19  

GENERAL   19  COMPLIANCE   19  

GLOSSARY  OF  TERMS   20  

APPENDIX  1   22  

APPENDIX  2   23  

APPENDIX  3   24  

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INTRODUCTION Companies and institutions worldwide have increased their understanding of the need of an effective exposure control program in order to minimize the occupational risks associated with bloodborne pathogens and Biological Safety Level 2 agents. While measures implemented and actions taken from institution to institution can vary severely depending upon risks and methods of exposure, the minimization of these biological risks can be met through an effective management system regardless of location. This system should continually improve measures to minimize biological risks through a cycle of effective planning, implementing, reviewing and improving actions to meet specified goals. This can also be referred to as the PDCA (Plan-Do-Check-Act) principle. Planning: Includes identification of hazards, risk and establishing goals Do: Implementing, including training and operational issues Check: Checking, including monitoring and corrective action Act: Reviewing, including process innovation and acting to make

needed changes to the management system By continually conducting PDCA assessments of biological hazards, risk of exposure is diminished while control is increased. The Dartmouth College Exposure Control Program (DC-ECP) has been developed to minimize employee exposure to bloodborne pathogens, (such as Hepatitis B and HIV) and agents classified by the CDC/NIH as requiring Biological Safety Level 2 (BSL2) practices and procedures. The DC-ECP is in compliance with the Occupational Safety and Health Administration’s (OSHA’s) Bloodborne Pathogen Standard (29CFR 1910.1030), and serves as both the written program for OSHA compliance and as a training document. Beyond blood borne pathogens, this policy discusses the College’s Biological Safety Program for BSL2 laboratories. Scope The Exposure Control Program applies to all principal investigators, laboratory personnel, students and visitors conducting research at a BSL2 level. The following information pertains to all laboratory, research and support activities that involve exposure to biological agents or materials at a BSL2 level with the goal of educating staff and personnel to the proper measures to ensure a safe research environment.

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Responsibilities It is the responsibility of the supervisors to ensure that all employees subject to occupational exposure follow the provisions of this plan. These responsibilities include but are not limited to providing a copy of this plan to employees, enforcing compliance with this document, ensuring new employees are properly trained, ensuring all employees attend an annual training session, and performing follow-up procedures for all exposure incidents. Personnel are to perform tasks and procedures in a manner that minimizes or eliminates employee exposure and perform duties as established in this exposure control plan and as they are trained. The Dartmouth Environmental Health and Safety Office provides OSHA-mandated bloodborne pathogen information and training sessions at least annually to each supervisor and employee with occupational exposure. It is the responsibility of each supervisor to ensure all pertaining personnel under his/her supervision complete this required training. DETERMINING RISKS OF EXPOSURE In the Dartmouth College research laboratories, the primary method of determining potential exposure is based on information gathered from an online Biological Research Registration Survey (https://dartmouth.bioraft.com/). Initial completion and a periodic update of this form by each Principal Investigator (PI) in all biomedical and biological laboratories are mandatory. The Office of Environmental Health and Safety on behalf of the College’s Institutional Biological Safety Committee (IBC) then review this information. If the use of animals is involved, an Animal Care and Use Form must be approved through the Institutional Animal Care and Use Committee (IACUC) and a current Biological Safety Registration Form must be on file. Planning for Risk Assessment of Exposure Supervisors and personnel should continually assess exposure risks, especially when agents, personnel, experimentation, location and/or instrumentation change according to protocol. To do so will minimize exposure and biological relevance of exposures. Planning of biological risk and identification should include all aspects of identifying external influences and hazards as well institutional changes to personnel training. Below is an example of one type of strategy for conducting a risk assessment.

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Other Risk Determination All research conducted at Biological Safety Level 3 (BSL3) must first be presented to the Biological Safety Officer for presentation to the Institutional Biosafety Committee (IBC). BSL3 research must meet IBC approval before commencement of research. Exposure determination for ancillary personnel will be conducted on a case-by-case basis. The document entitled Hazardous Waste Management describes methods of handling Medical Pathological Waste (MPW). These procedures help control and limit the potential for exposure in laboratories, housekeeping, maintenance and other support areas.

Taken from Guidance on CWA 15793:2008

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METHODS OF EXPOSURE CONTROL General Improved engineering and regulation of work practices are the primary means of elimination or minimization of exposure for personnel. Where occupational exposure remains after institution of these controls, personal protective equipment is to be used.

v Universal Precautions are used to prevent exposure to bodily fluids/tissues or other materials containing biological pathogens. These precautions are to be taken at all times when working with all BSL2 agents or when the risk of exposure/contamination is present.

v All clinical specimens of blood, human tissue and body fluids are to be

handled utilizing BSL2 work practices and procedures. (Please refer to the latest edition of the Biological Safety Level 2 (BSL2): Practices and Procedures in Research Laboratories Working with Human Blood and Body Fluids/Bloodborne Pathogens Resource Guide.) These practices, procedures and facility requirements are described in detail in the CDC/NIH publication entitled ”Biosafety in Microbiological and Biomedical Laboratories” (5th edition) and are to be followed by all Dartmouth College laboratories working with biological materials. A copy of this document is available from Environmental Health & Safety (EH&S). Each registered laboratory must have a copy of this document readily available in the lab at all times.

v Ancillary personnel are required to follow site-specific procedures developed

by EH&S. Please contact EH&S for details of these procedures. Posting Hazard Requirements v All labs working with BSL2 agents must post the EH&S provided BIOHAZARD

information sign on the outside door to the laboratory. This sign indicates the specific hazard, PI and emergency contact information. The information on the sign should be updated as circumstances change. The sign is readily available from EH&S.

v All labs working with human blood or body fluids/bloodborne pathogens must

post the “Essential Information on Laboratory Safety at Dartmouth College” flip chart. In this chart describes first aid needed in the event of an exposure to human blood and bodily fluids, biological personal protection, and waste decontamination and removal procedures.

v Any piece of equipment in a common room area (such as cold room,

incubator room, equipment room, ect.) that contains or is used with BSL2

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agents or may be contaminated with blood bodily fluids must have a biohazard warning label attached. The label must state the biological hazard, date and person of contact. These labels can be obtained from the Scientific Stockroom (Stock # 6934).

Engineering Hazard Controls Engineering controls are to be utilized where there is a likelihood of occupational exposure. v Primary Barriers. Class II Type A Biological Safety Cabinets (BSC) or other

physical containment devices must be used when procedures are performed with a potential for creating splashes or aerosols. Such procedures may include centrifuging, grinding, vortexing, blending, sonication and opening containers of infectious materials. Intranasal inoculations or other animal procedures that have the potential for producing splashes and aerosols must be performed in a biological safety cabinet. In special cases, procedures (such as an animal necropsy) may be performed on an open bench if it is determined by the Biological Safety Officer that the employee is at a significant increased risk of percutaneous exposure, to an infectious agent transmissible by the bloodborne route, when working in a biological safety cabinet. In these cases, strict adherence to mucous membrane protective practices is required.

v Annual Inspections of Biological Safety Cabinets. Annual inspection and

certification of biological safety cabinets is the responsibility of the primary investigator. EH&S maintains a database of biological safety cabinets and will aid in the scheduling of inspections and certifications. If a new cabinet is acquired or there are problems with old ones or if the lab is planning on moving a cabinet, please contact EH&S for assistance (646-1762).

v Mechanical Pipetting Devices. Mechanical pipetting devices are to be used

for all pipetting activities. Mouth pipetting is strictly prohibited. Aerosol-resistant pipette tips are recommended to control carry-over contamination caused by aerosol formation. The tips are ideal for a wide variety of applications including PCR, tissue culture, forensic studies, gel loading, and serological assays as well as pipetting radioactive samples

v Sharps Containers. Puncture resistant sharps containers are to be used at all

work sites where needles and syringes, Pasteur pipettes, scalpel blades and other sharps are used. Sharps containers should be closed and prepared for disposal when sharps are at ¾ of the full indicator line on the sharps container. Please refer to the Hazardous Waste Management, Minimization and Disposal Guide for the correct disposal option for your building. Puncture resistant sharps containers are available through the Scientific Stockrooms.

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v Safer Needle Devices. On April 18, 2001, the Occupational Safety and

Health Administration (OSHA) issued the Needlestick Safety and Prevention Act. Congress directed OSHA to make changes to its Bloodborne Pathogens Standard (CFR 1910.1030) to require the use of safer needle devices when drawing blood. EH&S will provide training on the use of these devices. For further information, please contact EH&S. You must be approved by EH&S in order to conduct blood drawing in your laboratory.

v Safety Devices for Centrifuges. For low speed centrifugation of infectious

materials, safety centrifuge cups must be used. If used, the cups are to be loaded and unloaded only within a biological safety cabinet. High-speed centrifugation of infectious material must be performed using a suitable gasketed rotor that is loaded and unloaded within a biological safety cabinet. Inside a biological safety cabinet, wipe off the exterior of the rotor with a suitable disinfectant before loading and unloading.

v Vacuum Line Protection. All vacuum lines must be protected by using in-line

vacuum filters and vacuum traps. These disposable filters are available through the Scientific Stockrooms. (Stock #5860) In labs where the vacuum lines are used routinely, it is recommended that the filters be changed every six months. In other areas, it is recommended that filters be changed on an annual basis. Immediately change the filter if the system no longer works effectively or becomes overtly contaminated.

v Vacuum trap decontamination. In an effort to readily decontaminate vacuum

trap waste upon use, vacuum trap disinfection measures must be taken. Appropriate levels of Wescodyne disinfectant for the vessel used must be added prior to vacuum trap use. The waste must be removed from the flask whenever ½ volume of flask volume is reached or every 3 months (which ever comes first). Vacuum trap decontamination schedule stickers must be placed on all vacuum trap flasks to serve as a record of proper disinfection and disposal. These stickers are readily available from EH&S as seen below.

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Work Practice Controls v Hand Washing. Hand washing is the single most important means of

preventing exposure to, or spread of, infectious agents. Hand washing must be performed after removing gloves, upon completion of a task within a biological safety cabinet, before leaving the lab for any reason, when hands are known or expected to be contaminated and before contact with face or mouth. Hands should be lathered thoroughly with an antimicrobial soap, vigorously washed for approximately 15-30 seconds then rinsed with copious amounts of water. Use paper towels to turn off the water faucet to avoid immediate recontamination.

v Food and Consumables. Eating, drinking, smoking, applying cosmetics or lip

balm, and handling contact lenses are prohibited in all laboratory settings and other work areas where there is a possibility of occupational exposure. Food and drink shall not be kept in laboratories or refrigerators, freezers, shelves, cabinets or on countertops or bench tops.

v Sharps Etiquette. Contaminated needles and other contaminated sharps shall

not be bent, recapped, or removed except when the lab supervisor determines that no alternative is feasible or that such action is required by a specific procedure. Recapping should only be performed using a mechanical recapping device. Shearing or breaking of contaminated needles is prohibited.

v Aerosols. All procedures liquid infectious material shall be performed in such

a manner as to minimize splashing, spraying, spattering, and the generation of droplets of these substances.

v Transport. The transport of blood or infectious material outside of the

laboratory will only be done in containers that prevent leakage. If outside contamination of the primary container is suspected, the primary container shall be placed within a second container that prevents leakage during handling, processing, storage, transport, or shipping. If the specimen could puncture the primary container, the primary container shall be placed within a secondary container that is puncture-resistant in addition to the above characteristics. For transport of the material outside of the facility additional requirements apply, contact the EH&S office for more information (646-1762).

Personal Protective Equipment v Laboratory Coats. Lab coats are the primary barrier between lab personnel

and infectious agents. Lab coats are to be worn at all times while working on, or adjacent to, all bench top procedures utilizing hazardous chemicals, biological or unsealed radiological materials. Laboratory coats must be appropriately sized for the individual and be fully buttoned when worn.

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Sleeves must be of appropriate length as to not expose skin while wearing gloves. Lab coats are not be taken from the laboratory and brought to any non-laboratory areas, especially where food and drink is being prepared and served. It is the responsibility of the PI to ensure all laboratory personnel adhere to Dartmouth’s standard of wearing laboratory coats.

v Gloves. Gloves are to be worn by all employees directly handling biological

material or contaminated surfaces. Vinyl examination gloves, surgical synthetic, or N-DEX® nitrile gloves may be chosen by the employee based on individual need and preference. Gloves are to be inspected before use and changed routinely. Gloves must be replaced when visibly soiled, torn, or punctured. All gloves must be discarded into an autoclave bag for proper disposal. Information on glove selection and chemical resistance is found on a poster displayed in each laboratory and from EH&S. Gloves are never a substitute for thorough hand washing. Hands should be thoroughly washed when entering the lab, after removing gloves and before leaving the lab.

v Mucous Membrane Protection. Mucous membranes must be protected by

wearing a surgical-type mask with safety glasses or a surgical-type mask with attached acetate eye shield when working with (viable) infectious materials outside of a biological safety cabinet. (FaceMask: Scientific Stockroom # 14342.)

v Safety Glasses/Shields. Safety glasses are recommended at all times in the

laboratory. v Personal Clothes. Any unprotected skin is forbidden. This means open toed

shoes, sandals and other open footwear as well as shorts or skirts that leave the skin unprotected is prohibited in all Dartmouth laboratories.

v Respirators. Respirators must not be used in the laboratory without prior

approval from EH&S. Supervisors are not authorized to select or recommend the use of respiratory protection, regardless of the type. Contact EH&S if you feel respiratory protection is required. Surgical facemasks used for mucous membrane protection are not considered respirators and are not to be used in situations where respiratory protection is required. All respirator users must be enrolled in the Dartmouth College Respiratory Protection Program. EH&S supplies and maintains the recommended respiratory protective devices.

See also Appendix 1: Personal Protective Equipment: Hazard Assessment and Use Requirements at Biological Safety Level 2.

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HOUSEKEEPING General Supervisors shall ensure that the laboratory and other work areas are maintained in a clean and sanitary condition. The lab supervisor shall determine and implement an appropriate schedule for cleaning and method of decontamination based upon the location within the facility, type of surface to be cleaned, type of soil present, and tasks or procedures being performed in the area. Decontamination and Spill Clean-up All work surfaces and equipment that come into contact with blood, bodily fluids and infectious agents must be disinfected immediately. Contaminated work surfaces shall be decontaminated with an appropriate disinfectant after completion of procedures; or as soon as possible when surfaces are contaminated, or at the end of the work shift if the surface may have become contaminated since the last cleaning. Spills within work areas are to be cleaned by laboratory or research personnel. Custodial staff are not authorized to clean up spills of biological material within lab areas. Protective coverings, such as plastic wrap, aluminum foil, or imperviously-backed absorbent paper used to cover equipment and environmental surfaces, shall be removed and replaced when they become overtly contaminated or at the end of the work shift if they may have become contaminated during the shift. All receptacles intended for reuse which have become contaminated with blood or other biologic materials shall be inspected and decontaminated immediately after use and upon visible contamination. Reusable sharps that are contaminated with blood or other biological materials shall not be stored or processed in a manner that can place employees at risk of suffering cutaneous punctures. SPILL CLEAN-UP PROCEDURE It is the responsibility of all personnel to know and understand the steps and procedures involved in spill clean-up of biological materials. If a spill is too large to be sufficiently contained and cleaned by laboratory personnel, please contact EH&S at 646-1762. For all other spills please refer to the following:

1. Alert persons in the immediate area that a spill has occurred. Stand away from the spilled material for 5 minutes to allow aerosols to settle.

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2. Wearing the appropriate protective equipment (gloves, lab coat, eye protection), cover the spill with paper towels.

3. Carefully pour a freshly prepared 1 in 10 dilution of household bleach

around the edges of the spill working to the center. For large spills, bleach may be added directly.

4. Allow a minimum 15 minute contact time.

5. Wipe and collect the spill by working from the edges to the center.

Broken glass should be carefully discarded into the appropriate container.

6. Broken glassware which may be contaminated should not be picked up directly with the hands. Use mechanical means, such as a brush and dust pan, tongs, or forceps. Don’t forget to decontaminate these mechanical devices after contact with the broken glass.

7. Clean the spill area again by wiping with fresh disinfectant.

8. Place all materials into an autoclave bag and follow the autoclave

instructions. In the event of a spill of biological material in a public access area (hallway, elevator, etc.), keep all personnel away from the spill and contact EH&S at 646-1762.

BIOLOGICAL WASTE DISPOSAL All employees must comply with the guidelines set forth in the Hazardous Waste Management, Minimization and Disposal Guide. Copies can be obtained by contacting EH&S at 646-1762. EMERGENCY STEPS TO TAKE IN THE EVENT OF BIOLOGICAL EXPOSURE In the event of an exposure to human blood or body fluids (bloodborne pathogens), follow these steps:

1. Immediately initiate first aid at the work site. o Contaminated skin should be thoroughly washed for 10 minutes using

a povidone iodine solution or an antiseptic soap and copious amounts of water.

See also Appendix 2: Performance Characteristic of Surface Disinfectants

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o Contaminated eyes and mucous membranes should be irrigated for 15 minutes with water.

2. Notify supervisor, if immediately available. Otherwise, go to step three. 3. Report to the Occupational Medicine at DHMC between 7:30 and 4pm.

Telephone: 650-7018. After hours, report to the DHMC emergency room. For emergency transport dial 911.

(DMS@ Borwell dial 5555. DMS @ Hanover dial 911.) MEDICAL SURVEILLANCE AND POST-EXPOSURE EVALUATION The Dartmouth College Exposure Consultation Program The purpose of this program is to provide risk assessment, medical consultation and follow up (when necessary) for members of the Dartmouth Community. This voluntary program is for individuals who have an elevated risk of exposure to hazardous substances in the workplace. Members of the Dartmouth Community who have concerns related to exposure to hazardous substances in the workplace should first contact EH&S. EH&S will conduct an initial workplace evaluation. Medical consultations (if needed) are coordinated through EH&S. EH&S will be given a written medical opinion but does not have access to individual medical records. Animal Exposure Surveillance Program The Occupational Health Program has an important role in the Dartmouth College Animal Care and Use Program. This program is operated by the Animal Resources Center (ARC) through the Department of Occupational Medicine, DHMC, and is designed to protect both DC-DMS personnel and the laboratory animals. This program consists of an informational handout that should be given to all personnel with animal contact and a program of periodic physical exams, tests, and vaccinations for personnel with an elevated risk of exposure. Elevated risk of exposure with laboratory animals is determined by a review of the Personnel Information sheet completed by the Principal Investigator on each individual in their laboratory working with animals. The level of participation in the Occupational Health Program is based on the species the individual is working with, the amount of time in contact with the animals and hazards identified in the protocol. The requirements of this program are based on guidelines in the NIH Guide for the Care and Use of Laboratory Animals. For more information on this program contact the Animal Resource Center http://dms.dartmouth.edu/arc/.

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Post-Exposure Evaluation Post-exposure evaluation will be performed under the direction of the Department of Occupational Medicine, DHMC. Hepatitis B Virus and HBV vaccination v Occupationally-acquired HBV. Hepatitis B infection is the leading

occupationally-acquired illness among health care workers, affecting approximately 15,000 workers annually. Hepatitis B virus (HBV), formerly know as “serum hepatitis” is one of several viruses which attacks the liver producing swelling, tenderness and sometimes permanent liver damage. HBV is spread primarily through contact with blood and body fluids that contain blood. The virus enters the body through open wounds, or breaks in the skin, needle sticks or other punctures or splashes of blood and/or body fluids to the mucous membranes.

v Symptoms of HBV. The most frequent symptoms of HBV infection include

fatigue, mild fever, muscle or joint pain, nausea, vomiting, loss of appetite and abdominal pain. Many symptoms suggest a flu-like illness but tend to last longer and jaundice may occur in up to 25% of cases. However, 50% of infected individuals have no symptoms.

v Risk of HBV Infection. Assuming BSL 2 practices and procedures are

followed, the risk of HBV infection in Dartmouth College employees is considered to be well controlled and limited even if their jobs entail frequent contact with blood and bodily fluids. Employees must be alert to situations where they, their co-workers or support staff may be exposed to blood and body fluids.

v HBV Vaccination. A recombinant HBV vaccine is available, free of charge, to

all Dartmouth College employees who have occupational exposures to blood and bodily fluids. At risk employees are strongly encouraged to become vaccinated.

Further information on hepatitis B and the HBV vaccine can be obtained by calling EH&S at 646-1762. EMPLOYEE NOTIFICATION Laboratories and other work areas that handle human blood and bodily fluids or any human pathogen must be posted with the BIOHAZARD information sign provided by EH&S (See Attachment 3). Any special requirements for entry into a workspace will be designated on the biohazard sign affixed to the entry door. All equipment that handles or processes hazardous biological materials must be labeled with a biohazard sticker to alert fellow personnel.

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RDNA REGISTRATION The National Institutes of Health (NIH) requires that laboratories working with rDNA be registered for certain types of rDNA experiments. It is the responsibility of each Principal Investigator (PI) at Dartmouth to ensure that their laboratory is in compliance with these requirements. The PI determines which research projects are exempt, which need to be registered simultaneous with initiation and which projects need prior registration and approval. If you are unsure which category your experiments fall into or if you need a copy of the NIH rDNA Guidelines, please visit: http://www.nih.gov/oba/ (To access the registration survey, go to https://dartmouth.bioraft.com/.) ENHANCED BIOLOGICAL SAFETY LEVEL 2 (BSL2) v Definition. Enhanced BSL2 is the practice of incorporating increased

personal protection equipment and procedures with a strict adherence to BSL2 practices and procedures in a setting which increases the possibility of occupational exposure to biological organisms (such as production of aerosols or high titer volumes of virus). For research involving high hazardous chemicals please contact EH&S and see Appendix F of the Chemical Hygiene Plan.

v Biological Organisms and Work Practices. Only research incorporating

biological organisms of BSL1 and BSL2 classification will be allowed at the Enhanced BSL2 level. No research involving BSL3 organisms will be allowed under Enhanced BSL2 conditions. Enhanced BSL2 signage shall be posted on the exterior lab door and laboratory access shall be strictly limited to direct lab users and Enhance BSL2 authorized personnel. All work involving infectious agents will be conducted inside a certified BSC with daily decontamination of solid/liquid waste and removal of all PPE before exiting laboratory.

v Authorization. The necessary precautions and practices for this biological

safety level will be determined on a case by case basis after a thorough risk assessment of the proposed research project performed by EH&S. To complete this risk assessment, a research specific safety manual must be written by the corresponding laboratory and submitted for approval through EH&S. This safety manual will discuss all aspects of the proposed research including agents used, authorized personnel, decontamination procedures, and emergency procedures. A copy of this safety manual must be kept in the

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laboratory at all times with easy access to Enhanced BSL2 personnel. All Enhanced BSL2 research must be cleared through EH&S and is subject to IBC approval before commencement of research. All Enhanced BSL2 personnel must be fully trained through EH&S at a BSL2 level.

For authorization of Enhanced BSL2 procedures please contact Dartmouth EH&S.

TRANSPORTATION OF BIOLOGICAL MATERIALS General When biological materials must be transported between campuses or via commercial services (nationally or internationally) such as UPS® or FedEx®, specific rules and requirements must be followed. When transporting materials on public roads or in the air, the requirements of the Department of Transportation (DOT) and International Air Transport Association (IATA) will apply. Shipping Categories The first step in determining how to ship your biological material is to select a category. The most common categories of biological materials offered for shipment from the College are Infectious substances (Category A & Category B) and Patient Specimens (animal and human). It is the shipper’s (laboratories) responsibility to determine the category of material you wish to ship and if an import or export permit is required for international shipments. The CDC website has information on import and export permitting. The country you are shipping to will also have specific import requirements which must be met. To determine which category your shipment falls into, use the Classification Flow Chart in Appendix 8. Contact EH&S to confirm you have correctly classified your material. v Category A. When shipping Category A infectious substances, certified

standard packaging is required as well as certification of training for the person offering the material for shipment. (Contact EH&S for this training or for assistance in the shipment) Proper certified packaging for infectious material may be purchased from companies such as Saf-T-Pak™. Infectious substances are considered “Dangerous Goods” and must be declared as such when shipping or transporting off campus. This is very important – failure to declare a dangerous good is illegal and could result in substantial fines and other penalties.

See also Appendix 3: MANUAL FOR RESEARCH CONDUCTED UNDER ENHANCED BSL2 CONDITIONS

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v Category B. When shipping Category B infectious substances, certified packaging is not required. Category B shipments require specific marking and labeling as a Biological substance, category B. Specific packaging details also apply.

v Patient Specimens. When shipping Patient Specimens, certified packaging is

not required and will be labeled as either Exempt Human Specimen or Exempt Animal Specimen.

v Transportation Between Campuses. When transporting biological material

between campuses, the following packaging is required: 1. A styrofoam container (outer package) with a tight fitting lid for

secondary containment. The lid must be taped securely shut during transport.

2. A biohazard label on the top and at least one side of the Styrofoam container, with the name of the material written on the label.

3. The primary container (inner package) must be durable and leak proof.

Resources Packing materials Saf-T-Pak : http://www.saftpak.com/

Import/Export information

CDC : http://www.cdc.gov/od/eaipp/

Shipping information

FedEx : http://fedex.com/us/service-guide/our-services/options/index.html?qgroup=toggle-c12&qid=FedEx_Declared_Value_Max_Liability

EQUIPMENT REPAIR AND TRANSFER All equipment which may have been exposed to hazardous materials (i.e., known hazardous chemicals, radiological or biological substances) must be appropriately decontaminated and tagged with a Dartmouth College Property Management tag prior to transfer, servicing or repair. (See instructions at the following url http://www.dartmouth.edu/~ehs/essential-info/hazard_clearance.html.) TRAINING AND EDUCATION Initial training of BSL2 practices and procedures is required for all personnel working in such facilities. BSL 2 training for laboratory personnel is provided on a regular basis. Primary investigators and supervisors are responsible for ensuring that all personnel under their direction receive training prior to handling biological materials and complete annual refresher training. Primary

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investigators and supervisors are responsible for job-specific safety training and must ensure that employees have been adequately trained or otherwise have the requisite knowledge/education to work safely. Monitor the EH&S Blitzmail Bulletin for upcoming training dates. Ancillary personnel will be given site-specific training according to the requirements specific to their work. INSTITUTIONAL BIOSAFETY COMMITTEE General Dartmouth College's Institutional Biological Safety Committee (IBC) provides oversight and review of the College’s biological safety program. It serves as an advisory board to the research community at Dartmouth on biosafety and recombinant DNA. Committee functions include those designated for the IBC in the NIH Guidelines for Research Involving Recombinant DNA Molecules. The IBC meets regularly and meetings are open to the public. Please contact Dartmouth’s Biological Safety Officer, Dr. Brian J. O’Shea, for additional information. Compliance It is the responsibility of the primary investigator to update their lab’s biological profile. This information will be used by the Biological Safety Officer to present to the IBC for institutional approval. To update your biological profile please log on to https://dartmouth.bioraft.com/

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GLOSSARY OF TERMS

Ancillary Personnel - non-laboratory staff who through the performance of their job duties may enter laboratories or come into contact with materials from research laboratories. They include, but are not limited to, personnel such as custodians, glassware washing facility personnel and building maintenance staff.

Blood – includes human blood, human blood components, or products made from human blood.

Bloodborne Pathogens - pathogenic microorganisms that are present in human blood and can cause disease in humans. These pathogens include, but are not limited to, hepatitis B virus (HBV) and human immunodeficiency virus (HIV).

Clinical Laboratory - a workplace where diagnostic or other screening procedures are performed on blood or other potentially infectious materials.

Contaminated - the presence or the anticipated presence of biologic materials on an item or surface.

Contaminated Laundry - laundry that has been soiled with blood or other potentially infectious materials or may contain sharps.

Contaminated Sharps - contaminated object that can penetrate the skin including, but not limited to, needles, scalpels, broken glass, broken capillary tubes, razor blades, etc.

Decontamination - the use of physical or chemical methods to remove biological pathogens on a surface or item to the point where they are no longer capable of transmitting infectious particles and the surface or item is rendered safe for handling, use, or disposal.

Engineering Controls - controls (e.g., sharps disposal containers, self-sheathing needles, safer medical devices, such as sharps with engineered sharps injury protections and needleless systems) that isolate or remove the biological hazard from the workplace.

Exposure Incident – an event in which potential for infections is heightened after coming into contact with biological pathogens

Handwashing Facilities - a location which provides an adequate supply of running potable water, soap and single use towels or hot air drying machines.

Occupational Exposure – an exposure incident that takes place during ones working duties.

July 2011 page 21

Personal Protective Equipment (PPE) - specialized clothing or equipment worn for protection against a specific hazard.

Sterilize - the use of a physical or chemical procedure to destroy all microbial life including highly resistant bacterial endospores.

Universal Precautions - an approach to infection control. According to the concept of Universal Precautions, all human blood and certain human body fluids are treated as if known to be infectious for HIV, HBV, and other bloodborne pathogens.

Work Practice Controls - controls that reduce the likelihood of exposure by altering the manner in which a task is performed (e.g., prohibiting recapping of needles).

July 2011 page 22

Appendix 1

Essential Information on Personal Protective Equipment: Hazard Assessment and Use Requirements at Biological Safety Level 2 The Occupational Safety and Health Administration (OSHA) requires that personal protective equipment (PPE) be selected, provided and worn by employees in situations where PPE could help reduce the potential for harm and injury (29 CFR 1910.132). This guide outlines the likely situations and hazards that require the use of PPE at Biological Safety Level 2 (BSL2). These requirements also apply to work done with animals at ABSL2 Special considerations may apply for certain tasks--contact EH&S whenever such questions arise. PPE is readily available to Dartmouth employees and students through the Scientific Stockrooms. Eye/face/mucous membrane protection (29 CFR 1910.133): When working with potentially infectious materials outside of a biological safety cabinet mucous membranes must be protected by wearing a surgical-type with safety glasses or a surgical-type mask with attached acetate eye splash guard. Safety glasses are recommended at all times in the laboratory and required when working with hazardous materials. All safety glasses must meet ANSI Z87-2003. Foot/skin protection (29 CFR 1910.136): Open toed shoes, sandals and other open footwear is prohibited in all Dartmouth laboratories. Additionally, shorts and other garments that leave skin unprotected are forbidden in Dartmouth laboratories using potentially infectious materials. Protective lab coats designated for lab use are to be worn while in the laboratory. This protective clothing is removed and left in the laboratory before leaving for non-laboratory areas (e.g., cafeterias, library and administrative offices.) Hand protection (29 CFR 1910.138): Appropriate gloves must be worn for activities involving potentially infectious materials. Gloves must also be worn when there is potential contact contaminated surfaces or equipment. For potentially infectious materials, disposable nitrile gloves are the best choice. Dispose of gloves when contaminated, when work with potentially infectious materials is completed and before leaving the laboratory. Never wash or reuse disposable gloves. Gloves are never a substitute for through hand washing when entering the lab, after removing gloves and before leaving the lab. Wash your hands for a minimum of 30-60 seconds with copious amounts of liquid soap and water. Other Reminders-- · Decontaminate work surfaces daily and after a spill involving any viable material. · Properly autoclave your waste. Do not allow waste to accumulate. This Certification of Tasks Requiring Personal Protective Equipment Prepared in Compliance with 29 CFR 1910.132

July 2011 page 23

Appendix 2

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July 2011 page 24

Appendix 3

MANUAL FOR RESEARCH CONDUCTED

UNDER ENHANCED BSL2 CONDITIONS

Date:__________Signature:_________________________________________ (Primary Investigator)

Date:__________Signature:_________________________________________ (Brian J. O’Shea, Ph.D., Biological Safety Officer)

Insert Research Laboratory

Insert Location

July 2011 page 25

SAFETY PRACTICES DURING ENHANCED BIOLOGICAL SAFETY LEVEL TWO (ENHANCED BSL2)

A. OVERVIEW Please give a detailed overview of the research project. Include purpose for research, strains used, reason for Enhanced BSL2 status, and time frame of research project.

B. LOCATION

Please describe the location where the Enhanced BSL2 research will be performed. Please include information pertaining to biological safety cabinets, freezers, incubators, autoclaves, and any other equipment related to Enhanced BSL2 research.

C. ACCESS RESTRICTIONS

Access to areas of the lab being currently used for ENHANCED BSL2 research is restricted to lab members with ENHANCED BSL2 training. No glasswasher will handle ENHANCED BSL2 waste. No glasswasher, custodian, or maintenance staff will be allowed to interact with the biosafety cabinet or researcher at the biosafety cabinet when ENHANCED BSL2 work is underway.

D. ADMINISTRATION

1. ENHANCED BSL2 Facility Supervisors:

Insert Location

Insert Research Laboratory

Insert Text Here

Insert Text Here

July 2011 page 26

Please list researcher supervisors (PI and lab manager) with contact information for each.

2. Institutional oversight and approval: Work involving Enhanced

BSL2 practices requires previous approval through the Institutional Biosafety Committee (IBC) before commencement of research.

3. Written Biosafety Procedures: The laboratory keeps documents

describing the safe lab practices and procedures, developed in consultation with the EH&S office and IBC policies. All personnel involved in research in this lab are required to adhere to these written rules. These documents can be found on the Dartmouth EH&S website for Biological Safety. A copy of this document is to be kept for reference in the ENHANCED BSL2 laboratory in 314W as well as with the EH&S Office.

4. Responsibilities: The Dartmouth EH&S Office, on behalf of the IBC,

has the right and responsibility to review, inspect and approve the use of the location mentioned above for ENHANCED BSL2 work as described in this document.

The Principal Investigator is responsible for ensuring that before beginning ENHANCED BSL2 work, all personnel demonstrate proficiency in standard microbiological practices and receive appropriate training on the potential hazards associated with the work involved and the operations specific to the facility. It is the responsibility of the principal investigators to ensure compliance and supervise project personnel. The Dartmouth EH&S office serves as the administrative arm of the IBC and as such oversees the biosafety program at Dartmouth College. Their responsibilities include reviewing and presenting projects to the IBC; advising faculty and staff on proper biosafety procedures for the agents being handled and reviewing and approving the written procedures for all involved researchers and associated personnel. EH&S will also perform periodic inspections of the facility for compliance with the practices outlined in this document and relevant regulations and guidelines.

All personnel conducting ENHANCED BSL2 work recognize the risks inherent in working with human pathogens, and agree to take responsibility for adhering to appropriate protective measures as outlined in institutional guidelines and in this manual.

Insert Text Here Brian O’Shea, Ph.D. / Biological Safety Officer / 646-9790

July 2011 page 27

E. INFECTIOUS AGENTS

1. Modes of Transmission: Please describe the mode of transmission of the biological agents used in this study

2. Signs and Symptoms of Infection:

Please describe the signs and symptoms of clinical human infection.

3. Methods of Treatment:

Please describe the current method of treatment for infected individuals for the agents used in this study.

4. Methods of Disinfection:

Please describe the methods for disinfection of solids and liquids that will be used in this study.

F. LABORATORY FACILITIES AND SAFETY EQUIPMENT. 1. Biological Safety Cabinet: A properly maintained and certified

biological safety cabinet is to be used for all manipulations of ENHANCED BSL2 materials.

No ENHANCED BSL2 work is to be conducted on the open bench top. This

includes opening containers of potentially infectious materials, pipetting, dilutions, transfer operations, plating, framing, grinding, blending, drying,

Insert Text Here

Insert Text Here

Insert Text Here

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July 2011 page 28

sonicating, and shaking. ENHANCED BSL2 materials can be centrifuged using centrifuge safety cups or sealed rotor heads with transfer into them using an uncontaminated sealed container.

A labeled spray bottle of 70% ethanol and a container of 10% bleach are

kept at each biosafety cabinet for decontamination before and after each work session.

2. Equipment for Treatment, Disposal and Handling of Wastes:

Appropriate equipment for treatment, disposal and handling of wastes (disinfectants, aspirator flasks, autoclavable waste containers, biohazard red-orange bags, sharp collectors, etc.) shall be readily available inside the ENHANCED BSL2 facility, and are to be used according to procedures described below.

Please describe the location of the autoclave to be used for these

experiments.

3. Cell Culture Incubators: ENHANCED BSL2 materials can be kept in

cell culture incubators in a closed container. The cell culture incubator is to be cleaned on a scheduled basis.

4. Water bath. ENHANCED BSL2 materials can be thawed in the water

bath if they are kept in a sealed and water tight container. 5. Additional Equipment: Small quantities of ENHANCED BSL2

material in closed containers can be examined or processed in the microscope, refrigerator, freezer or microcentrifuge.

6. Table-Top Centrifuge: For centrifugation of ENHANCED BSL2

material, screw capped tubes and aerosol-containing canisters must be used, as well capped centrifuge buckets. If there has been a spill in the hood or concern exists that the outsides of containers to be centrifuged are contaminated, the outsides must also be wiped down with 70% ethanol before removing from the biosafety cabinet. If there has been any possibility of leakage during centrifugation, the inner walls of the centrifuge and the rotor will be immediately decontaminated. Leakages during centrifugation must be reported immediately to the Principal Investigator.

Careful attention must be given to placing a balanced load into the centrifuge. Balance problems will create experimental problems and will increase the possibility of tube breakage. Centrifuge and centrifuge

Insert Text Here

July 2011 page 29

buckets and walls are to be cleaned on a scheduled basis. When a spill of any type occurs, the centrifuge must be cleaned immediately and before any continued use. See appendix for cleaning protocol.

7. Pipetting Aids: When ENHANCED BSL2 material is pipetted using

disposable pipettes in the biosafety hood, care should be taken to avoid aerosolization. In addition, after pipetting is complete, bleach decontamination of pipettes will be undertaken such as through pipetting the diluted bleach waste solution up and down in the pippetor prior to pipette disposal in a pipette coffin. Care should be taken that liquid is not drawn in the pipette aid. This can be prevented by not over-filling the pipette. If liquid enters the pipette aid it will be collected in the filter enclosed in the aid. You will know this has occurred as the pipette will no longer aspirate or dispense liquid. If this happens, remove the pipette from the unit. Remove the nose cone area of the aid by using a twisting action. Remove the contaminated filter and discard in a biohazard waste container. Use a tissue wetted with alcohol to wipe the inside area before replacing the filter. This is an acrodisc filter with a pore size of 0.45um.

8. Laboratory Doors: A large biohazard sign is present on the door to

the area in which ENHANCED BSL2 work occurs. The door shall be self-closing and shall have a glass window for sight capability into the room. A red emergency notification card listing the names and phone numbers of the contact persons is also posted.

9. Sink: The lab shall contain a sink that is to be used for the disposal of

liquid ENHANCED BSL2 waste after proper chemical decontamination, as described below. Bottles of concentrated bleach are stored underneath the sink. Soap and paper towels are available from dispensers near the sink. An eyewash/drench hose is located at a nearby sink located in the laboratory.

10. Vacuum Lines are protected with secondary traps and in-line

hydrophobic filters. 11. Facility: The lab is designed so that it can be easily cleaned and shall

be maintained free of trip hazards. 12. Bench Tops: No ENHANCED BSL2 work will be conducted on bench

tops. 12. Emergency Eyewash: An eye wash hoses and safety showers shall

be made readily available. Please list the locations of the eye wash and safety showers for the

research laboratory in question.

July 2011 page 30

13. Personal Protective Equipment: When ENHANCED BSL2 work is

performed, researchers will wear disposal reusable biosafety gowns and gloves.

a. Gowns. All disposable lab gowns will be disposed of with ENHANCED BSL2 waste when contaminated or in need of replacement. Disposable gowns and soiled lab coats will not be worn outside of the lab. b. Gloves. Gloves are disposed of when contaminated, replaced frequently, and removed when work with potentially infectious materials is completed or when leaving the area. Gloves are not worn in the common areas of the lab where ENHANCED BSL2 work is not done. Gloves are not to be used to handle common equipment such as phones, desks, and door knobs. Disposable gloves are not washed or reused. Do not spray gloves with alcohol prior to working. Gloves that fit over the sleeves of the lab gown are recommended. c. Face protection. Face protection (glasses or a faceshield) is used whenever splashes, spatters, or sprays of infectious or other hazardous materials may be generated and contamination to the face (eye, nose, or mouth) can be anticipated.

14. Labeling: Equipment used for ENHANCED BSL2 work must be

labeled with a universal biohazard symbol and marked ENHANCED BSL2. Labeling should include name, date, and the identity of the material.

15. The EH&S Office must be notified of any changes in the procedures or

practices contained in this document. G. LABORATORY PRACTICES

1. Immunizations: It is recommended that those working with human blood, body fluids, tissues, etc. receive the Hepatitis B vaccination. This will be provided free of charge. All laboratory personnel, whether they have direct or indirect contact with Enhanced BSL2 agents, should consult with EH&S and Occupational Medicine regarding possible vaccinations. Information on receiving the vaccine will be provided at the time of BSL2 training. Contact EH&S if you wish to consult with Occupational Medicine.

Insert Text Here

July 2011 page 31

2. Laboratory Doors: Doors are kept closed at all times and locked when laboratory is unattended. 3. Training: The PI is responsible for ensuring that all lab personnel receive appropriate training before conducting ENHANCED BSL2 work. This includes standard EH&S training plus review of this manual. Personnel are required to read and follow the instructions on practices and procedures in this document. 4. Pre and Post-Work Cleanup Procedures: The procedures for pre and post- ENHANCED BSL2 work cleanup include the following: Pre-Work Procedures 1. Clean the work surface with 70% ethanol or 10% bleach. 2. Spray off all equipment and tools required for work with 70% ethanol. 3. Place all equipment and tools required for work inside the hood. 4. If a sharps container is required, place it inside the hood. 5. Waste container(s) is available and lined with a biohazard bag. 6. Check that the biosafety cabinet is functioning properly. 7. Check that vacuum flask is prepared with Wedscodyne and lines are intact. 8. Dress in all required personal protective equipment. Post-Work Procedures 1. Place all contaminated materials in biohazard bags. 2. Clean with 70% ethanol all equipment removed from cabinet. 3. Clean hood with 70% ethanol or 10% bleach.

4. In the event of potential aerosolization, clean glass sash inside and outside with 70% ethanol.

5. Remove personal protective equipment taking care to keep from spreading contamination.

6. Store or discard personal protective equipment. Disposable gowns shall be placed in labeled biohazard bags and stored in designated area. 7. Wash Hands. 5. Use of Biological Safety Cabinet: All work with open containers with ENHANCED BSL2 material must be performed in the biological safety cabinet. The biosafety cabinet blower will remain ON when ENHANCED BSL2 work is being done in the cabinet. Decontamination of all biological waste must be performed with pure bleach before removal from biological safety cabinet. All biohazardous materials will be transferred out from the biosafety cabinet only in a leakproof container which has been tightly closed and, if contaminated, decontaminated by spraying all over the outsides with 70% ethanol and/or 10% bleach.

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Activities that may disrupt the inward directional airflow through the sash opening of a biosafety cabinet are to be minimized. In particular, repeated insertions and withdrawal of the worker’s arms in and out from the work chamber, improper placement of materials, blockage of the front or rear grilles, or brisk walking past the cabinet while it is in use. Work inside the biosafety cabinet should be performed at least four inches behind the front air intake grille and arms should not rest on the grille. All personnel are responsible for thoroughly cleaning the biosafety cabinet with 70% ethanol and/or 10% bleach after each procedure is completed. 6. Packaging: Cultures, tissues, or specimens of body fluids are placed in sturdy containers that prevent leakage during collection, handling, processing, storage, or transport. The shipment and off-campus transport of biohazardous materials is strictly regulated by the US Dept. of Transportation. The EH&S office maintains packaging supplies and expertise for shipping biohazardous materials. Please contact EH&S well in advance of any planned shipments or transport of biohazardous materials off site. 7. Transport of ENHANCED BSL2 material: The transport of ENHANCED BSL2 material will be done only in sealed, biohazard labeled containers such as a tupperware box. No transport of unsealed containers which contain ENHANCED BSL2 material are allowed in public spaces. ENHANCED BSL2 materials may not be transported to another lab or research area that is not registered and approved for ENHANCED BSL2 work. Any exceptions require EH&S and/or IBC approval. 8. Sharps: No sharps are used when conducting ENHANCED BSL2 work if this is at all avoidable. Instead, use plastic lab ware and avoid scalpels, needles or other such equipment. As a general rule, with any work with human tissue, sharps are avoided and a high degree of precaution must always be taken with any contaminated sharp items, such as broken glass, slides, capillary tubes, and pipettes. 9. Spills: All spills must be cleaned up immediately and only by trained properly equipped scientific personnel. Autoclave all toweling and sponges used in the cleanup and wash your hands thoroughly afterwards.

a. Spills inside the biosafety cabinet: Immediately stop work and remove hands/arms from BSC after decontaminating any areas that came into contact with spill material. If spill material came into contact with gloves, remove gloves in BSC and leave inside BSC. A 10 minute wait is recommended after leaving BSC in order for any aerosolized material to be filtered by BSC HEPA filters before returning to BSC for spill clean-up. Chemical decontamination shall be performed while the cabinet continues to operate. Remove contaminated gloves and put on a clean pair. Spray and

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wipe the walls, work surface, and equipment with disinfectant such as 70% ethanol or freshly diluted Clorox with a 10% final concentration. The drain pans and catch basins below the work surface of a Class II cabinet may need to be flooded with disinfectant for a 20 minute contact time before draining.

b. Large Spills outside the biosafety cabinet: Report immediately to

the principal investigator and the EH&S Office after evacuating the room. If an aerosol risk exists, wait 30 minutes for dissipation before reentering the room to complete the emergency procedures. Medical evaluation and appropriate follow-up will be provided by the Occupational Employee Health Department (650-7018) for any exposure incident. Written records will be maintained.

c. Equipment Contamination: Label as contaminated and then wearing

protective gear, isolate any leaky container into a plastic bag and decontaminate the spill and all contaminated surfaces following the above procedures.

Equipment shall also be decontaminated before it is sent for repair or for

maintenance or moved to another lab area. d. Reusable Labware: Must be completely submerged in a freshly

prepared 10% Bleach solution for at least 20 minutes and rinsed with water before taking out of the lab or moving to the BSL2 lab space and being given to the glasswashing staff. This includes tissue culture media bottles, beakers, and any nondisposable pipets. Currently very little reusable labware is used.

10. Accidents Affecting Laboratory Personnel: Immediate emergency

procedures have to be followed in case of accidents that result in overt exposures of personnel to infectious materials. In case of a splash into an eye, nose or mucous membranes of the mouth, the eyewash located by the sink in 314W will be used immediately. In case of skin exposure, thorough washing with disinfectant soap and water at the ENHANCED BSL2 sink is recommended. In case of parenteral injection (e.g., puncture), forced bleeding is first recommended followed by thorough washing.

Accidents and all spills must be reported immediately to the Principal

Investigator and/or lab supervisor. Injured personnel must report for immediate medical evaluation, treatment, and post exposure follow up through the Occupational Employee Health Department (650-7018) between the hours of 7:30am-4:00pm. After hours treatment can be received at the DHMC emergency room (911 or x5555) as appropriate, and written records will be kept. An Accident Report Form must be submitted by the PI to the Office of Risk Management.

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11. Treatment of Contaminated Wastes: All ENHANCED BSL2 waste in

the biosafety cabinet must be identifiable as such and be kept separate from the regular BSL2 and BL1 waste. All ENHANCED BSL2 waste will be removed and treated at the end of each procedure by the scientist. In general, contaminated waste will not be left overnight.

ENHANCED BSL2 wastes and contaminated items will be decontaminated

by the scientist before being turned over to custodians or glasswash personnel. All liquid or solid wastes are decontaminated before disposal according to the Dartmouth College EH&S Hazardous Waste Management, Minimization and Disposal Guide.

a. Liquid Wastes: All liquid wastes generated during ENHANCED BSL2

experiments will be immediately decontaminated inside the biosafety cabinet by mixing with bleach (20% final concentration) for at least 20 minutes contact time. The solution may then be disposed of in the ENHANCED BSL2 sink. The sink must then be decontaminated and washed after the disposal.

Liquids aspirated under vacuum will be collected in a 1 liter aspirator bottle

which is placed inside a plastic bucket and initially contains approximately 10 ml of Wescodyne. The aspirator bottle is to be emptied when the volume is not more than 500 ml and replaced by the researchers at least every three months. A secondary bottle and a HEPA filter will be placed between the vacuum source and the primary aspirator bottle.

b. Non-Sharp Solid Wastes: All solid wastes contaminated with

ENHANCED BSL2 materials must be packed, tagged, transported, recorded, and autoclaved by trained scientists. A leakproof pan will be used to hold the biowaste bags or boxes ready for autoclaving. The logs and charts for each ENHANCED BSL2 waste load will be filed in a designated notebook. Autoclaving will be performed according to validation parameters. Autoclaves will be validated by the scientists for proper temperature and pressure control on a periodic basis. Contact EH&S for validation procedures. If an autoclave is not operating properly, a sign will be posted and the Physical Plant Director will be notified immediately.

Solid ENHANCED BSL2 waste (paper, plastic wrappers, gloves, plastic

plates, emptied plastic flasks and falcon tubes) will be collected inside the biosafety cabinet in a small, red bag inside a ENHANCED BSL2 labeled, autoclavable container such as a Nalgene plastic beaker or covered pail. The bag will be twisted closed inside the biosafety cabinet, removed directly into a second red bag, tagged with a numbered, white biowaste label, placed in a leakproof autoclave pan and then removed from the

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ENHANCED BSL2 facility and autoclaved by the scientist at the completion of the procedure.

All biohazard bags containing ENHANCED BSL2 contaminated materials

must be closed before being removed from the ENHANCED BSL2 facility and shall be transported to the autoclave in a secondary leakproof container or a metal autoclave pan; red bags of ENHANCED BSL2 waste will not be carried unprotected in the hallway. It is also preferable to transport materials using a cart. The autoclave log must be filled out for each run and the chart paper saved. See Autoclave log in the Appendix. These records are maintained for 3 years. The scientists are responsible for autoclaving their waste immediately after their experiment.. Once the solid waste is decontaminated, it will be placed inside a regular trash bag and be treated as ordinary trash by the housekeeping staff.

c. Sharps: Plastic pipettes and syringes fitted with filters used during

ENHANCED BSL2 work will be put into a coffin containing of freshly diluted 10% Clorox solution in the biosafety cabinet. The Clorox solution will be drawn into the entire length of the pipette or syringe. After at least 20 min of soaking, the pipettes and syringes will be carefully drained in the sink. The pipettes will be discarded into a small red bag with the rest of the BSL2 waste and disposed of accordingly.

12. Aerosols and Pipetting: Pipetting by mouth is prohibited.

Mechanical pipetting devices are to be used. Pipettes contaminated by non-ENHANCED BSL2 materials will be disposed of through the standard laboratory waste. Pipettes contaminated with ENHANCED BSL2 materials shall be washed with a bleach-containing solution after use.

13. Handwashing and gloves: ENHANCED BSL2 work requires double-

gloving. The outside glove must be removed before leaving the hood and disposed of in the bag for contaminated solid wastes. The inside glove must be removed and disposed of as with all biohazard trash. All personnel must wash their hands after conducting ENHANCED BSL2 work. Gloves are not to be used in handling common equipment such as telephones or incubator doors.

14. Food, etc.: Eating, drinking, smoking, handling contact lenses, taking

medications, and applying cosmetics are not permitted in the lab area.

15. Housekeeping: Cleaning and decontamination of work surfaces (benches, BSC, incubator, centrifuge, etc.) will be performed by the scientists, as described in institutional BSL2 policies. When ENHANCED BSL2 work is in progress, custodial and maintenance staff will not be given access to the hood or other areas in which the experiment is being

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conducted. Otherwise, service and maintenance staff should conduct themselves according to institutional policy.