CHAPTER 9 TABLE OF CONTENTS

Transcription

1 CHAPTER 9 TABLE OF CONTENTS 1.0 RESPONSIBILITIES INTRODUCTION ENVIRONMENTAL HEALTH AND SAFETY BIOLOGICAL AND CHEMICAL SAFETY PROGRAM LABORATORY MANAGEMENT STAFF/STUDENTS INSTITUTIONAL BIOLOGICAL SAFETY COMMITTEE (IBC) GENERAL GUIDELINES FOR HANDLING BIOLOGICAL AGENTS THE SEVEN BASIC RULES OF BIOSAFETY PERSONAL HYGIENE CLOTHING (PRIMARY BARRIERS) AND PERSONAL PROTECTIVE EQUIPMENT HANDLING PROCEDURES HANDWASHING PROTOCOL FOR HANDWASHING SYRINGES CONTROLLING THE BIOHAZARD AREA HOUSEKEEPING CONTAINMENT BIOSAFETY LEVELS UNIVERSAL PRECAUTIONS/STANDARD PROCEDURES GLOVE USE 3.1 INTRODUCTION GENERAL RECOMMENDATIONS Revised December 2015 Page 1 of 106

2 3.3 GLOVE MATERIALS AND TYPES GLOVE SELECTION SHIPMENT OF BIOLOGICAL AGENTS 4.1 OVERVIEW TRANSPORTATION AND TRANSFER OF BIOLOGICAL AGENTS 5.1 INTRODUCTION TRANSPORTATION REGULATIONS GENERAL PACKAGING REQUIREMENTS FOR TRANSPORT OF BIOLOGICAL AGENTS AND CLINICAL SHIPMENT TRANSPORTATION AND TRANSFER OF BIOLOGICAL MATERIAL ON CAMPUS SHIPMENT OF SELECT AGENTS: INTERNAL TRANSFERS OF SELECT AGENTS AEROSOLS 6.1 INTRODUCTION RISKS AEROSOL PRODUCTION MINIMIZING AEROSOL PRODUCTION DISINFECTION AND STERILIZATION 7.1 DEFINITIONS WASTE DISPOSAL GENERAL PROCEDURES CHOOSING A METHOD SELECTING A DISINFECTANT STERILIZATION METHODS Revised December 2015 Page 2 of 106

3 7.7. AUTOCLAVING PROCEDURE FOR BIOLOGICAL WASTE BIOLOGICAL SAFETY CABINETS 8.1 OVERVIEW PRIMARY CONTAINMENT: BIOLOGICAL SAFETY CABINETS CLASS I CLASS II CLASS III POSITIVE-PRESSURE PERSONNEL SUIT OTHER DEVICES TABLE1- COMPARISON OF BIOLOGICAL SAFETY CABINETS CLASS I BIOLOGICAL SAFETY CABINET CLASS II, TYPE A1 BIOLOGICAL SAFETY CABINET TABLE TOP MODEL OF A CLASS II, TYPE A2 BIOLOGICAL SAFETY CABINET CLASS II, TYPE B1 BIOLOGICAL SAFETY CABINET (CLASSIC DESIGN) CLASS II, TYPE B2 BIOLOGICAL SAFETY CABINET CLASS III BIOLOGICAL SAFETY CABINET CERTIFICATION SAFETY RULES FOR USE OF CLASS I AND II CABINETS UNDERSTANDING THE MONITORS STARTING THE JOB COMPLETION OF A JOB REMOVAL OF MATERIAL FROM THE BIOSAFETY CABINET BIOHAZARDOUS AND RDNA SPILLS IN THE CABINET REMEMBER DO S AND DON TS OF THE BIOSAFETY CABINET MAINTENANCE OF THE BIOSAFETY CABINET UV LIGHTS IN BSC PROCEDURES FOR CENTRIFUGATION 9.1 LOW-SPEED CENTRIFUGATION MICROFUGE USE Revised December 2015 Page 3 of 106

4 9.3 BIOHAZARDOUS AND RDNA CENTRIFUGE SPILL DECONTAMINATION PROCEDURES PROCEDURES FOR SPILLS OF BIOHAZARDOUS & RDNA MATERIALS 10.1 SPILLS IN THE LABORATORY IN CASE OF A SPILL DECONTAMINATION AND CLEAN-UP PROCEDURES SPILLS IN HALLWAYS RE-OCCUPANCY OF A SPILL AREA PROCEDURES FOR OCCUPATIONAL EXPOSURE INCIDENTS RECOMBINANT DNA 11.1 DEFINITION RESEARCH APPROVAL CATEGORIES NIH RECOMBINANT GUIDELINES RECOMBINANT DNA ROLES AND RESPONSIBILITIES THE 5 NIH CATEGORIES IBC NOTIFICATION INFORMATION NEEDED FINK COMMITTEE REPORT RISK ASSESSMENT ADDITIONAL INFORMATION DUAL USE NOTIFICATION OF USE FOR BIOLOGICAL AGENETS AND RECOMBINANT DNA 12.1 INTRODUCTION SELECT AGENT USE CHANGES TO THE SELECT AGENT REGISTRATION Revised December 2015 Page 4 of 106

5 12.4 SELECT AGENT LIST LABORATORY MOVES/TRANSFERS/CLOSURES 13.1 GENERAL LABORATORY EQUIPMENT DECONTAMINATION FORM EQUIPMENT RESPIRATORY PROTECTION PROGRAM 14.1 OVERVIEW INTRODUCTION TERATOGEN EXPOSURE HAZARDS KNOWN HUMAN TERATOGENS TRAINING REQUIREMENTS INSTITUTIONAL BIOSAFETY COMMITTEE INTRODUCTION BIOLOGICAL SAFETY COMMITTEE SCOPE AND MISSION STATEMENT DEFINITION OF A BIOLOGICAL AGENT REVIEW CLASSIFICATION REVIEW PROCEDURES APPEALS EXAMPLES OF NOU AND INSTRUCTIONS APPENDICES APPENDIX A - ANIMAL VIRAL ETIOLOGIC AGENTS IN COMMON USE APPENDIX B - PATHOGENS OF VETERINARY SIGNIFICANCE APPENDIX C - FOOTNOTES AND REFERENCES APPENDIX D: LABORATORY BIOSAFETY LEVEL CRITERIA TABLE 1. SUMMARY OF RECOMMENDED BIOSAFETY LEVELS FOR INFECTIOUS AGENTS Revised December 2015 Page 5 of 106

6 19.5 APPENDIX E: VERTEBRATE ANIMAL BIOSAFETY LEVEL CRITERIA TABLE 1. SUMMARY OF RECOMMENDED BIOSAFETY LEVELS FOR ACTIVITIES IN WHICH EXPERIMENTALLY OR NATURALLY INFECTED VERTEBRATE ANIMALS ARE USED APPENDIX F - BIOSAFETY IN MICROBIOLOGICAL AND BIOMEDICAL LABORATORIES (BMBL) APPENDIX G - BLOODBORNE PATHOGEN EXPOSURE CONTROL POLICY REFERENCES Revised December 2015 Page 6 of 106

7 1.0 RESPONSIBILITIES 1.1 Introduction The National Institutes of Health (NIH) and Centers of Disease Control and Prevention (CDC) defines biohazards (biological hazards) as "infectious agents presenting a risk or potential risk to the well-being of man, or other animals, either directly through infection or indirectly through disruption of the environment." Proper handling and disposal of biohazardous materials greatly reduces the potential for exposure to infectious or harmful agents. General information and guidelines are presented in this chapter. Details for the safe use of specific biological agents and rdna may be obtained from Environmental Health and Safety, Biological and Chemical Safety. 1.2 Environmental Health and Safety Biological and Chemical Safety Program Environmental Health and Safety Biological and Chemical Safety Program (EHS-B&C) will: provide consultation and technical information for handling biological agents review proposals and protocols (Notification of Use, NOU) for the use of human products, hazardous biological agents, rdna, human gene therapy, and submit these to the Institutional BioSafety Committee, and provide the Institutional Biological Safety Committee with recommendations oversee registration of Select Agent users and laboratories with the CDC receiving and shipping of Select Agents present biological safety seminars upon request oversee the annual certification of biological safety cabinets by an external contractor provide weekly autoclave efficacy testing for waste decontamination autoclaves review and approve purchases of biological safety cabinets and other safety-related equipment survey laboratories for compliance with approved standards and policies of UTMB, CDC and NIH, and other governing agencies provide assistance or advice in the disinfection of facilities and equipment assist in the development of safety plans and training programs BioSafety Officer will notify agencies of incidents and accidents as appropriate to local, state and federal regulations and guidelines including but not limited to CDC/DSAT, NIH-OBA and USDA/APHIS. BioSafety Officer will notify IBC of incidents and accidents that require reporting to Federal Agencies including but not limited to NIH-OBA, USDA/APHIS and CDC/DSAT. 1.3 Laboratory Management Management will: observe the established guidelines and polices for biological safety Revised December 2015 Page 7 of 106

8 ensure that laboratory personnel are trained in the hazards and safe handling procedures of biological agents encourage employees to report any changes or suspected changes in their health status advise Environmental Health and Safety - Biological and Chemical Safety of any significant changes in laboratory protocol for the use of biological agents report all biological and rdna spills and incidents to EHS Biological and Chemical Safety Program. 1.4 Staff/Students Staff/Students will: observe the established guidelines and policies for biological safety inform immediate supervisor of any unsafe practices or conditions in the work area report to supervisor any change or suspected change in their health status if there is a possibility it may be work-related report all biological and rdna spills and accidents to their supervisor 1.5 Institutional Biological Safety Committee (IBC) The Institutional Biological Safety Committee will: establish general policies and standards for the safe use of biologicals at UTMB define categories of biohazardous agents and rdna to determine if specific policies are needed for their safe use review Notification of Use (NOU) applications involving biological agents and recombinant DNA/RNA molecules approve, recommend changes, or deny the use of the biological agents or rdna/rna molecules ensure that there are no undue hazards to the participants, other staff members, or the public verify the adequacy of the technical procedures performed in order to minimize occupational exposure review the qualifications of the applicant undertaking the proposed research review and report to NIH OBA any rdna incident deemed to be significant. 2.0 General Guidelines for Handling Biological Agents 2.1 The Seven Basic Rules of Biosafety The most common means of exposure can be essentially eliminated as occupational hazards by following these seven basic rules of biosafety. Do not mouth pipette. Manipulate infectious fluids carefully to avoid spills and the production of aerosols and/or droplets. Revised December 2015 Page 8 of 106

9 Restrict the use of needles and syringes to those procedures for which there are no alternatives; use disposable or needle-locking syringes to avoid self-inoculation; and dispose of sharps in leak and puncture-resistance containers. Use protective laboratory coat, gloves, and eye protection. Wash hands following all laboratory activities, or contact with infectious materials/animals. Decontaminate work surfaces before and after use, and immediately after spills. Do not eat, drink, store food, or apply cosmetics in the laboratory. These procedures are targeted at minimizing overt occupational exposures and constitute basic essentials of good laboratory practice. 2.2 Personal Hygiene Whenever handling biological material, wash your hands thoroughly: after working with any biohazards or animals after removing gloves, lab coat, and other contaminated protective clothing before eating, drinking, smoking, or applying cosmetics before leaving the laboratory area You should always keep your hands away from your face while handling biological material. Do not: eat drink smoke apply cosmetics in the work area 2.3 Clothing (Primary Barriers) and Personal Protective Equipment Clothing: clothes should be comfortable and practical for the type of work conducted. No open toe shoes will be worn in the lab. Hair will be properly held back as to not require to be brushed back by the hand Gloves and Lab Coat: are NOT worn outside the laboratory area. BSL 1 It is recommended that lab coats or gowns be worn to prevent soiling of street clothes. Water proof bandage and double gloves will be worn if skin on hands is broken or if a rash is present. Gloves must be worn to protect hands from exposure to hazardous materials. Protective eyewear (if splashes are anticipated) should be worn. Use of contact lenses is not recommended; if contact lenses are used, laboratory eye protection or face protection must be worn. BSL 2 Protective laboratory coats, gowns, smocks or uniforms designated for lab use are worn when in the laboratory. Revised December 2015 Page 9 of 106

10 Gloves are worn when hands may contact potentially infectious materials, contaminated surfaces or equipment. Double gloving is recommended when handling infectious material. Face protection is used for anticipated splashes or sprays of infectious or other hazardous materials when biohazardous material is manipulated outside the Biological Safety Cabinet (BSC). BSL 3 Solid front or wrap-around gowns are used when in the lab; re-usable gowns are decontaminated (autoclaved) prior leaving the lab to be laundered. Disposable gowns are autoclaved prior to leaving the lab. Gowns are changed when contaminated or damaged. Double gloves must be worn when handling infectious materials, infected animals and contaminated equipment, two pairs of gloves is highly recommended. Respiratory and face protection are used in rooms containing infected animals or if determined by the Institutional Biosafety Committee. 2.4 Handling Procedures Always Use mechanical pipetting devices and cotton-plugged pipettes. Perform all procedures with a minimum of aerosol production. Add a disinfectant to water baths used with infectious substances. Use trunnion cups with screw caps or sealed rotors for centrifuging procedures. If safety cups/sealed rotors are not available, run centrifuge inside BSC. Inspect centrifuge tubes for cracks. Use secondary leak-proof containers when transporting samples, cultures, inoculated plates, or other containers of biohazardous materials. Place all containers on a lab cart for transport between laboratories. Label containers indicating contents, date and name of researcher. 2.5 Handwashing Handwashing is an extremely important procedure for preventing exposure to and dissemination of infectious agents. Unless microbial contamination is routinely removed, exposure via contact with mucous membranes, inoculation through skin, or ingestion becomes inevitable. Laboratory personnel should wash their hands: when coming on duty on leaving the laboratory for whatever reason when hands are obviously soiled or after cleaning a spill before and after completion of a task in a biological safety cabinet, even if gloves are worn before touching the face or mouth when removing gloves for any reason upon completion of duty 2.6 Protocol for Handwashing Revised December 2015 Page 10 of 106

11 A protocol for handwashing is as follows: Turn on faucets and wet hands with tepid water. Dispense non-antiseptic soap or antiseptic compound into a cupped hand. Spread soap or compound around both hands and between fingers. If needed, add a little more water to facilitate spread and lathering. Wash hands for about 30 seconds. Vigorously rub both sides of hands starting from a few inches above the wrist, extending downward between the fingers and around and under the fingernails. Rinse thoroughly under the tepid running water. Rinsing should start above the wrist area and proceed to the tips of the fingers. Note: if faucets are not knee- or foot-operated, do not turn off water (do not touch faucet handles) yet. Dry hands thoroughly with paper towels. If faucets are hand operated, turn them off now, using a dry paper towel to protect clean hands. 2.7 Syringes Avoid using syringes and needles whenever possible. If a syringe is necessary: use the needle-locking type, or a disposable syringe-needle unit place disposable syringes directly into sharp container non-disposable sharps & Luer-lock (glass) syringes must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving do not place syringes in pans containing pipettes or other glassware requiring sorting do not recap needles dispose of needles in leak proof, puncture resistant containers specifically designed for sharps disposal and in accordance with Chapter 10 - Disposal of Hazardous Materials 2.8 Controlling the Biohazard Area Keep laboratory doors closed. Laboratory doors are locked when lab is unoccupied. Limit access into the laboratory during procedures involving biohazardous agents. A sign will be posted that includes the universal biohazard symbol when infectious materials or infected animals are present in the laboratory or animal room. This warning sign must identify the agent and indicate requirements for entry (such as immunizations or respiratory protection), emergency contact information, and the approved biosafety level for the laboratory. Have a HEPA filter and vacuum trap will be used on laboratory vacuum lines. 2.9 Housekeeping Decontaminate work surfaces: -daily and -after each spill of biological material using appropriate disinfectant Decontaminate all potentially contaminated equipment used with an experiment. To decontaminate or sterilize materials at a site away from the laboratory, transport in a closed leak-proof container. Revised December 2015 Page 11 of 106

12 Dispose of contaminated wastes according to UTMB Policy for recycle and disposal of chemicals and biohazardous material. (See section Waste Disposal ). Keep books and journals only in clean areas of the laboratory. All equipment must be completely decontaminated prior to sending the equipment for routine maintenance or repair work Containment Containment defines the safe methods for controlling infectious agents where they are being handled. The purpose of containment is to reduce exposure to, and prevent the escape into the environment of, potentially hazardous agents. The three elements of containment include laboratory practice and technique, safety equipment, and facility design. Laboratory practice and techniques include the following: Strict adherence to standard microbiological practices and procedures (Biosafety in Microbiological and Biomedical Laboratories) Awareness of any potential hazards and training and proficiency in the practices and techniques associated with the materials being handled Use of appropriate safety equipment for the specific procedure The laboratory supervisor must be trained in laboratory techniques, safety procedures, and hazards associated with handling potentially infectious agents. Safety equipment includes primary barriers between the infectious agent and the worker includes biological safety cabinets, safety centrifuge cups, personal protective clothing is most effective when used with good laboratory techniques Facility design provides secondary barrier against potential exposure includes engineering features allowing protection of laboratory personnel, the work area, or the environment while handling hazardous materials most effective when combined with good laboratory technique and safety equipment 2.11 Biosafety Levels Four biosafety levels are described by the Centers for Disease Control and Prevention (CDC) and the National Institutes of Health (NIH) to recommend laboratory practices, safety equipment, and facilities appropriate for the potential hazards posed by the laboratory activity and the microorganism involved. The practices, safety equipment, and facility design for each biosafety level and animal biosafety level are fully described in the U.S. Department of Health and Human Services s Biosafety in Microbiological and Biomedical Laboratories Manual (See Appendix C of BMBL 5 th Ed). Biosafety Level 1 - practices, safety equipment, and facilities are appropriate for undergraduate and secondary educational training and teaching laboratories, and for other facilities in which work is done with defined and characterized strains of viable microorganisms not known to cause disease in healthy adult humans. Revised December 2015 Page 12 of 106

13 Biosafety Level 2 - practices, equipment, and facilities are applicable to clinical, diagnostic, teaching and other facilities in which work is done with the broad spectrum of indigenous moderate-risk agents present in the community and associated with human disease of varying severity. With good microbiological techniques, these agents can be used safely in activities conducted on the open bench, provided the potential for producing splashes or aerosols is low. Biosafety Level 2 is appropriate when work is done with any human-derived blood, body fluids, or tissues where the presence of an infectious agent may be unknown. Primary hazards to personnel working with these agents relate to accidental percutaneous or mucous membrane exposures, or ingestion of infectious materials. Biosafety Level 3 - practices, safety equipment, and facilities are applicable to clinical, diagnostic, teaching, research, or production facilities in which work is done with indigenous or exotic agents with a potential for respiratory transmission, and which may cause serious and potentially lethal infection. Primary hazards to personnel working with these agents relate to auto inoculation, ingestion, and exposure to infectious aerosols. Biosafety Level 4 - practices, safety equipment, and facilities are applicable for work with dangerous and exotic agents, which pose a high individual risk of life-threatening disease, which may be transmitted via the aerosol route, and for which there is no available vaccine or therapy. Additionally, agents with a close or identical antigenic relationship to Biosafety Level 4 agents should also be handled at this level. The primary hazards to personnel working with Biosafety Level 4 agents are respiratory exposure to infectious aerosols, mucous membrane exposure to infectious droplets, and auto inoculation Universal Precautions/Standard procedures Laboratories in healthcare and research facilities may handle human specimens. Human blood, blood products, human tissue and biological body fluids (urine, feces etc.) are to be handled as infectious or potentially infectious. Specimens may contain multiple infectious etiologic agents. To minimize personal exposure to specimens of an unknown nature, all personnel in laboratories will observe Centers for Disease Control (CDC) guidelines for universal/standard precautions when handling all specimens of tissue, blood and body fluid. This means that all human material will be considered to be infectious and will be handled as potentially hazardous. (Appendix G) 3.0 Glove Use 3.1 Introduction Gloves are an integral piece of personal protective equipment (PPE) at UTMB. Gloves are used in research, healthcare, animal, grounds keeping and mechanical areas. Attention to glove type and glove material is essential to providing protection for the intended use. No single glove type or material is universally appropriate. 3.2 General Recommendations Gloves should be worn when handing hazardous biological material or when protection of the biological material from contamination is required. Gloves chosen should be of a material known to be resistant to permeation by the agent for the duration of use, while allowing sufficient dexterity. Revised December 2015 Page 13 of 106

14 Inspect gloves for discoloration, small holes or tears before use. Remember that all waterproof gloves will develop pinholes over time. Remove gloves before touching objects such as doorknobs, elevator button, telephones or computers. Reusable gloves should be washed and inspected before and after each use. Remove gloves and wash hands before leaving the laboratory. Note: Wearing the wrong type of glove could be more hazardous than wearing no gloves by: giving a false sense of protection holding the hazardous agent in prolonged contact with the skin creating new hazards by decreasing dexterity Gloves are not to be worn outside the laboratory. Containers/equipments are to be placed inside a disinfected closed container and marked appropriately prior to leaving the laboratory 3.3 Glove Materials and Types The most common glove material used when handling biologicals are: latex - powdered - powder free polyvinyl chloride (PVC) nitrile 3.4 Glove Selection When selecting gloves one should consider the type of work and type of hazard. Important parameters for gloves for physical protection of the hands include material strength, dexterity, permeation, abrasion, and heat or cold resistance. When working with biologicals it is important to also consider the chemicals that are being used in order to choose the correct glove material for both biological and chemical protection. This may involve gloves changes or double-gloving to provide the best protection for the particular job. Problems with latex gloves include reaction and/or allergies to both the powder and the latex material itself. Gloves are available that are powderless. Contact Environmental Health and Safety for glove selection and report suspected allergic reactions to UTMB Employee Health Center. The following factors should be considered when selecting gloves: Chemicals will ultimately penetrate a glove, and may do so without evidence of damage to the material. Although gloves may protect against a biological hazard, they may not protect against a chemical or a mixture of chemicals. Higher temperatures than room temperature decrease break-though time of most materials. Thicker materials or multiple layers are usually better for combined biological and chemical use. Contaminated gloves must be discarded after use. Surgical gloves, in general, must meet strict quality standards and may provide a higher level of protection when working with more hazardous agents/tasks. Revised December 2015 Page 14 of 106

15 Note: Disposable gloves are designed for single use. Do NOT reuse these gloves. Dispose of them in the appropriate container after use. Additional information regarding gloves and chemical use is found in the UTMB, Chemical Safety chapter. 4.0 Shipment of Biological Agents 4.1 Overview The transportation of hazardous agents is strictly regulated. Failure to adhere to applicable regulations can result in fines and/or punitive actions against the university and the transporter. In addition to violating state and federal transportation laws, personal liabilities can be associated with failure to follow the appropriate shipping and handling requirements. The U. S. Department of Transportation (US-DOT) regulations regarding the shipment of hazardous materials state that no person may offer or accept a hazardous material for transportation in commerce unless the hazardous material is properly classed, described, packaged, marked, labeled and in condition of shipment. (HM-171.2). The International Air Transport Association (IATA) Dangerous Goods Regulations were developed so that dangerous goods can be transported safely by air transport throughout the world. It is necessary for all employees involved in the preparation or transport of dangerous goods, to include both diagnostic specimens and infectious substances, to be properly trained. No UTMB employee is permitted to ship regulated diagnostic and infectious substances without having completed certified DOT/IATA training every 2 years. EHS will be responsible for shipping of ALL Select Agents on campus. UTMB has a system to meet DOT and IATA requirements. Contact Environmental Health and Safety for information regarding training prior to shipment. EHS provides access to computer based training as well as sponsors in house training. The employee must successfully complete the training course in order to carry out these responsibilities. Please direct questions to EHS at extension Contact UTMB Technology Management to ensure all international shipping requirements are in place when planning to ship an item outside of the United States. U.S. Department of Commerce Export licenses may be required and may take several months to arrange. Also the receiving country may have import license requirements that must be arranged prior to shipment. 5.0 Transportation and Transfer of Biological Agents 5.1 Introduction Biological agents include infectious agents of humans, plants, and animals, as well as toxins that may be produced by microbes or by genetic material potentially hazardous by itself or when introduced into a suitable vector. Etiologic agents and infectious substances are closely related terms that are found in the transfer and transportation regulations. Biological agents may exist as purified and concentrated cultures but may also be present in a variety of materials such as body fluids, tissues, soil samples, etc. Biological agents and the materials that are known or suspected Revised December 2015 Page 15 of 106

16 to contain them are recognized by federal and state governments as hazardous materials and their transportation and transfer is subject to regulatory control. Transportation refers to the packaging and shipping of these materials by air, land, or sea, generally by a commercial conveyance. Transfer refers to the process of exchanging these materials between facilities. 5.2 Transportation Regulations on the transportation of biological agents are aimed at ensuring that the public and the workers in the transportation chain are protected from exposure to any agent that might be in the package. Protection is achieved through (a) rigorous packaging that will withstand rough handling and contain all liquid material within the package without leakage, (b) appropriate labeling of the package with the biohazard symbol and other labels to alert the workers in the transportation chain to the hazardous contents of the package, c) documentation of the hazardous contents of the package should such information be necessary in an emergency situation, and (d) training of workers in the transportation chain to familiarize them with the hazardous contents so as to be able to respond to emergency situations. Transportation of select agents is strictly regulated. No individual laboratory is allowed to receive or ship select agent material directly into the laboratory. 5.3 Regulations US Public Health Service (USPHS) 42 CFR Part 72. Interstate Transportation of Etiologic Agents. This regulation is in revision to harmonize it with the other U.S. and international regulations. A copy of the current regulation may be obtained from the Internet at: Department of Transportation. 49 CFR Parts Hazardous Materials Regulations. This regulation applies to the shipment of both biological agents and clinical specimens. Information may be obtained from the Internet at: United States Postal Service (USPS). 39 CFR Part 111. Mailability of Etiologic Agents. Codified in the Domestic Mail Manual : Etiologic Agent Preparations. A copy of the Domestic Mail Manual may be obtained from the Government Printing Office by calling or from the Internet at: Occupational Health and Safety Administration (OSHA). 29 CFR Part Occupational Exposure to Bloodborne Pathogens. This standard provides minimal packaging and labeling requirements for transport of blood and body fluids within the laboratory and outside of it. Information may be obtained from your local OSHA office or from the Internet: or STANDARDS&p_id=10051 Revised December 2015 Page 16 of 106

17 Dangerous Goods Regulations (DGR). International Air Transport Association (IATA). These regulations provide packaging and labeling requirements for infectious substances and materials, as well as clinical specimens that have a low probability of containing an infectious substance. These are the regulations followed by the airlines. These regulations are derived from the Committee of Experts on the Transport of Dangerous Goods, United Nations Secretariat, and the Technical Instructions for the Transport of Dangerous Goods by air, which is provided by the International Civil Aviation Organization (ICAO). A copy of the DGR may be obtained by calling or through the Internet at: or General Packaging Requirements for Transport of Biological Agents and Clinical Specimens Annex 3 (Page 23) shows the generalized "triple" (primary receptacle, water tight secondary packaging, and durable outer packaging) packaging required for a biological agent of human disease or materials that are known or suspected of containing them. This packaging requires the "Infectious Substance" label shown in Figure 2 on the outside of the package. This packaging must be certified to meet rigorous performance tests as outlined in the USDOT, USPS, USPHS, and IATA regulations. Clinical specimens with a low probability of containing an infectious agent are also required to be "triple" packaged, but performance tests require only that the package shall not leak after a four-foot drop test. 5.5 Shipment Regulations on the shipment of biological agents are aimed at ensuring that the change in possession of biological materials is within the best interests of the public and the nation. These regulations require documentation of the personnel, facilities, and justification of need for the biological agent in the shipment and subsequent approval of the transfer process by a federal authority. The following regulations fit in this category: Importation of Etiologic Agents of Human Disease The Centers for Disease Control and Prevention s Import Permit Program (IPP) regulates the importation of infectious biological agents, infectious substances, and vectors of human disease into the United States. Prior to issuing an import permit, IPP reviews all applications to ensure that entities have appropriate safety measures in place for working safely with these imported materials. Inspecting Permittees IPP may inspect applicants to ensure that the facilities have implemented the appropriate biosafety measures for the infectious biological agent, infectious substance, or vector to be imported. nt_or_vector.pdf Guidance for completing the CDC PHS import permit can be found on the internet at: Revised December 2015 Page 17 of 106

18 You can also to: Importation of Animal and Plant Pathogens United States Department of Agriculture (USDA), Animal and Plant Health Inspection Service (APHIS) permits are required for infectious agents of livestock and biological materials containing animal material. Tissue culture materials and suspensions of cell culture grown viruses or other etiologic agents containing growth stimulants of bovine or other livestock origins are controlled by the USDA due to the potential risk of introduction of exotic animal diseases into the U.S. Further information may be obtained by calling the USDA/APHIS at (301) (see ). U.S. Fish and Wildlife Service permits are required for certain live animals, including bats. Please call WILD for further information ( ). Importation of Plant Biological Agents 7 CFR Part 330. Federal Plant Pest Regulations; General; Plant Pests; Soil; Stone and Quarry Products; Garbage. This regulation requires a permit to import or domestically transfer a plant pest, plant biological agent, or any material that might contain them. Information can be obtained by calling or through the Internet at: Transfer of Select Biological Agents of Human Disease 42 CFR Part Transferring or Receiving Select Agents. Facilities transferring or receiving select agents must be registered with the CDC and each transfer of a select agent must be documented. Information may be obtained on the Internet at: Export of Etiologic Agents of Humans, Animals, Plants and Related Materials Exports of Infectious Materials The export of a wide variety of etiologic agents of human, plant, and animal diseases may require a license from the Department of Commerce. Information may be obtained by calling the Department of Commerce Bureau of Export Administration at or through the internet at: Contact UTMB Technology Management group for help with information on export licenses and Material Transfer Agreement. Shipment Revised December 2015 Page 18 of 106

19 For further information on any provision of this regulation contact: Centers for Disease Control and Prevention Attn: External Activities Program Mail Stop F Clifton Road N.E. Atlanta, GA Telephone: (404) FAX: (404) For further information on packaging and shipping of a biological material contact: Environmental Health and Safety Biological and Chemical Safety Transportation and transfer of biological material on campus. Transport of biological agents on campus, between building, within building, requires that the person transporting the material has knowledge of the agent/material being moved, how to respond to a spill if dropped, and has been provided with appropriate training in the packaging of the agent/material being transported. All efforts will be made to prevent a spill or aerosol. The outer container needs to be properly surface decontaminated and the agent will never be left unattended while in transit between laboratories. The biohazard level of the agent needs to be respected; you may not take a biohazard agent in a laboratory that is of lower containment specification even if the secondary container is not opened in that laboratory (e.g. BSL3 agents may not be brought into a BSL2 laboratory). Biological material must be triple packed, primary container that has agent is to be placed inside a leak proof secondary container (this can be a zip sealed plastic bag, a screw top conical tube, a pressure sealed plastic box (i.e. Rubbermaid container) which is then placed in a carrier. The carrier must be labeled with the name of the principal investigator/supervisor responsible for the laboratory and must have a biohazard label when moving a biohazardous agent. The carrier must such that if dropped it will not break or come apart. Transportation of agents on campus is not regulated but needs to be documented and records kept in the laboratories Shipment of Select Agents: Transfer of select agent material is regulated and must be approved by EHS, laboratories receiving select agent material are approved to do so and follow the proper regulation of select agent possession. Note: Shipments must be packed by a DOT/IATA trained and certified person. Revised December 2015 Page 19 of 106

20 Select agents will be shipped on a Monday, Tuesday or Wednesday only. Shipments are sent and received by EHS Shipments will not occur during the winter holiday season when the University is closed. Blackout dates will vary. Contact EHS before arranging shipments in or out of campus. 5.8 Internal transfers of Select Agents All internal transfers of select agents will be approved the Responsible Official/Alternate Responsible Official. Date and time of transfer will be arranged beforehand and the transfer witnessed by Biological and Chemical Safety. Revised December 2015 Page 20 of 106

21 Classification Flowchart Revised December 2015 Page 21 of 106

22 Revised December 2015 Page 22 of 106

23 Example of Packing and Marking for Category A Infectious Substances (See Packing Instruction 602 for additional requirements) Notes: Revised December 2015 Page 23 of 106

24 Example of Packing and Marking for Category B Infectious Substances (See Packing Instruction 650 for additional requirements, e.g. drop test) Notes: Revised December 2015 Page 24 of 106

25 Example of Packing and Marking for Exempt Specimens Notes: Revised December 2015 Page 25 of 106

26 6.0 Aerosols 6.1 Introduction The word "aerosol" refers to the physical state of liquid or solid particles suspended in air. The production of aerosols while handling infectious agents may present a serious risk of exposure. 6.2 Risks Aerosol particles one to five microns in size presents the greatest hazard to the laboratory worker because: Small particles readily penetrate and remain in the respiratory tract if inhaled. Many routine laboratory procedures create aerosols in this size range. They may remain suspended in air for long periods of time. Aerosols can settle on equipment normally considered to be clean. Skin contamination from aerosols or from handling contaminated equipment may result in infection through ingestion, mucosal membrane contact, or skin abrasions. It is also possible for aerosol particles to be spread by the building ventilation system. Risks associated with aerosols can be reduced or eliminated by the use of good technique in a biological safety cabinet. 6.3 Aerosol Production Aerosols may be produced in the use of: centrifuge blender shaker magnetic stirrer sonicator pipette vortex mixer syringe and needle freeze-dried sample vacuum-sealed ampoule grinder, mortar, and pestle test tubes and culture tubes heated inoculating loop separatory funnel lyophilizer 6.4 Minimizing Aerosol Production To reduce/minimize aerosol production: Revised December 2015 Page 26 of 106

27 perform activities that may produce aerosols in a biological safety cabinet (preferred) or a chemical fume hood keep tubes stoppered when vortexing or centrifuging allow aerosols to settle for one to five minutes (BSL1/2) and five to ten minutes (BSL3/4) before opening centrifuge, blender, or tubes that have been mixed place a cloth soaked with disinfectant next to the work area to deactivate possible spills or droplets of biohazardous agents reconstitute or dilute contents of an ampule slowly when mixing two solutions, discharge the secondary fluid down the side of the container or as close as possible to the surface of the primary solution allow inoculating loop or needle to cool before touching biological specimens wrap with disinfectant-soaked gauze when: -removing the needle from the rubber stopper of a test tube or vial, -breaking the cap on an ampule, -removing stoppers or plugs from tubes, or -expelling air or surplus solution from a syringe if the safety cups and/or rotors are not gasketed, run them in the biological safety cabinet (BSL1/2); Open centrifuge cups/rotors in the biological safety cabinet (BSL3/4) Do not: mix a solution by flushing with a pipette or syringe; MOUTH pipette; use a hypodermic and syringe as a substitute for mechanical pipetting devices when transferring infectious fluids. 7.0 Disinfection and Sterilization 7.1 Definitions Decontamination the application of microbiocidal steam, gas, solid (granular) or liquid chemical agents in situations in which microbes may be protected from contact by extraneous matter. Decontamination implies the destruction of or removal of microorganisms to some lower level, but not necessarily total destruction. Sterilization, disinfection and antisepsis are forms of decontamination Sterilization the total destruction of all living organisms (including spores) by processing in steam sterilizers (autoclaves), with ethylene oxide autoclaves or by chemical (high level) sterilization. Disinfection to destroy all non-spore forming organisms that could pose a potential hazard to humans or compromise the integrity of the equipment. Disinfection implies the use of antimicrobial agents on inanimate objects (floors, bench tops, equipment). Antisepsis the application of a liquid antimicrobial chemical to living tissue either human or animal. The objective is to prevent sepsis by either destroying potentially infectious organism or inhibiting their growth and multiplication. Revised December 2015 Page 27 of 106

28 7.2 Waste Disposal Laboratory waste needs to be segregated according to it nature and means of disposal. It is important to follow the proper disposal means for safety reasons but also in a concern for the cost of disposal and the threat to the environment. Please try to keep the waste generated to a minimal amount. Normal non hazardous waste (clear bags) This type of waste is handled by housekeeping and should contain non hazardous material such as paper, clean cell culture disposable plastic wear, non infectious nucleic acid and molecular biology disposable plastic wear and biological waste that has been chemically disinfected and autoclaved (pans from biological hoods containing bleach). Biohazardous waste to be autoclaved (red bags) This type of waste is generated by BSL2 laboratories and should be in red bags placed in a secondary container. The bags are picked up by housekeeping and are then taken to a centralized area on campus for autoclaving and disposal. Hazardous waste autoclaved prior to leaving the lab and then autoclaved in centralized place on campus before disposal. This type of waste is generated by high and maximum containment laboratories, for which all waste has to be autoclaved prior to leaving the facility. These are bags that come in a different color, but make sure they are autoclave approved or they will melt in the autoclave, causing extensive damage to the unit. Once this waste is out of the facility it is placed in a secondary container and is picked up by housekeeping and taken to a centralized area on campus for another autoclave cycle followed by disposal. Hazardous material to be incinerated (Yellow bags). This type of waste is to be placed in yellow bags and is very limited to certain research laboratories and chemotherapy areas. Approval for such means of disposal must be obtained by EHS. 7.3 General Procedures Disinfect frequently all floors, cabinet tops, and equipment where biohazardous materials are used. Decontaminate all infectious materials and contaminated equipment prior to being washed, stored, or discarded. Use autoclavable or disposable materials whenever possible. Keep reusable and disposable items separate. Minimize the amount of materials and equipment present when working with infectious agents. Decontaminate, properly store, or dispose of all biohazardous materials at the end of each day. Be aware that agar and other materials may interfere with the germicidal actions of chemical disinfectants, thus requiring pre-cleaning, higher concentrations, or longer contact time. Ensure sterilization by using suitable indicators with each autoclave load. Use clearly marked holding containers such as "NON-INFECTIOUS" or "BIOHAZARDOUS TO BE AUTOCLAVED". Revised December 2015 Page 28 of 106

29 7.4 Choosing a Method The method of choice for sterilization or disinfection will depend on two factors: the target organism (the biological agent) and the characteristic(s) of the materials or areas to be decontaminated. 7.5 Selecting a Disinfectant Use the following table to aid in the selection of an appropriate disinfectant. Substance Alcohols Phenols Quaternary Ammonium Compounds Chlorine Bleach Sodium Hydroxide DESCRIPTIONS OF COMMONLY USED DISINFECTANTS Description Ethyl or isopropyl alcohol at 70-80% concentration is a good general purpose disinfectant, not effective against bacterial spores. At BSL2/3/4, primarily used for control of environmental contamination. Not recommended for use with blood borne pathogens. Effective against vegetative bacteria, fungi and lipid-containing viruses; unpleasant odor; toxic by skin contact. Cationic detergents, which are strongly, surface active; extremely effective against lipoviruses; not effective against bacterial spores; may be neutralized by anionic detergents (soaps). Low concentrations (10%) active against vegetative bacteria and most viruses; higher concentration (50%) required for bacterial spores; corrosive to metal surfaces; must be prepared fresh; laundry bleach (5.25% chlorine may be diluted and used as a disinfectant). Sodium hydroxide at 1-2N for 1-2h can be used to disinfect material contaminated with Prions (BMBL) 7.6 Sterilization Methods Wet Heat (Steam) This method requires approximately 15psi pressure with a chamber temperature of at least 250 F (121 C). The cycle time begins when the materials being sterilized reach the predetermined temperature. Then the length of time is dependent upon the volume size of the load (usually minutes). Monitor steam sterilization effectiveness with a biological indicator, Geobacillus stearothermophilus. Revised December 2015 Page 29 of 106

30 Dry Heat This is less effective than steam, and requires more time (two to four hours) and a higher temperature ( F or C). Monitor dry heat sterilization with a Bacillus subtilis biological indicator. Biological Test Packs EHS monitors autoclaves used for waste decontamination. Geobacillus stearothermophilus. This process utilizes spores of Ethylene Oxide Gas (EO) Ethylene oxide sterilization in no longer available at UTMB. Contact EHS Biological and Chemical Safety for additional information if needed Autoclaving procedure for biological waste All biological waste that requires autoclaving should always be placed in a secondary container when being transported to and from the autoclave for decontamination. Waste bag that require autoclaving shall be double bagged and placed in a container for autoclaving. This will prevent spills in the autoclave. Bottles with liquids will also be placed in a secondary container prior to being autoclaved. Autoclave tape should be placed on the bottle to verify proper sterilization. Waste contained in trays will be placed in a secondary container labeled and autoclaved. DO NOT over fill trays with waste. The tray must be closed and all material should be immersed in disinfecting liquid. Use caution when removing items from autoclave. Steam and hot liquids may seriously burn you. Autoclave gloves face shields, and rubber aprons are available for added protection. Before disposing of the autoclave waste read the printout of the run to assure proper sterilization time, temperature, pressure. If an error is noted the waste must be re-autoclaved and facility maintenance should be notified. 8.0 Biological Safety Cabinets 8.1 Overview A biological safety cabinet is used as a primary barrier against exposure to infectious agents. A PRIMARY BARRIER SUCH AS A BIOLOGICAL SAFETY CABINET MERELY COMPLEMENTS CAREFUL WORK PRACTICES. Revised December 2015 Page 30 of 106