Laboratory BiosafetyLevel Criteria-Biosafety Level 2 suitable BoeateyLeval2issmaroBoaboleel1ads personnel and he environment.It differs from BSL-1 in that (1) laboraonYersonehgYvaeectcntncgnher2nacatoanenic the laboratory s limited when work is being conducted:(3) extreme precautions are taken with contaminated sharp items: and thes ma cabinets or other physical containment equipm ent. 8gentandacmesappyoagensas2gnegSoay The follow standard and special pra A.Standard Microbiological Practices 1 are in progress. 2 Persons wash their hands after the handle viable mate Eating.drinking,smoking,handling contact lenses,and apply 9.cosmetics not permitte in th treas refrigerators designated for this purpose only. 4. ogtDegingispohibiedmechanicalppetingdevie 5. Policies for the safe handling of sharps are instituted 6 All procedures are perfommed carefully to minimize the creation of splashes or aerosols 7. Work surfaces are decontaminated on completion of work or at the end of the day and after any spill or splash 21
Laboratory Biosafety Level Criteria – Biosafety Level 2 21 Biosa fety Lev el 2 is sim ilar to Biosafety Le vel 1 a nd is suitable fo r work inv olving age nts of m oderate potential ha zard to personnel and the environment. It differs from BSL-1 in that (1) labor atory p erso nnel have spec ific tra ining in hand ling pa thog enic agents and are directed b y com petent sc ientists; (2) ac cess to the laboratory is limited when work is being conducted; (3) extreme precautions are taken with contaminated sharp items; and (4) certain procedures in which infectious aerosols or splashe s ma y be create d are co nducte d in biologica l safety cabinets or other p hysical con tainme nt equipm ent. The fo llowing stan dard an d spec ial practices , safety equipm ent, and fa cilities apply to age nts ass igned to Biosafety Level 2: A. Standard Microbiological Practices 1. Access to the laboratory is limited or restricted at the discretion of the labo ratory directo r when e xperim ents are in progress. 2. Persons wash their hands after they handle viable materials, after removing gloves, and before leaving the labor atory. 3. Eating, drinking, smoking, handling contact lenses, and applying co sme tics are no t perm itted in the wo rk area s. Food is stored outside the work area in cabinets or refrigerato rs desig nated fo r this purpo se only. 4. Mouth pipetting is prohibited; mechanical pipetting devices are used. 5. Policies for the safe handling of sharps are instituted. 6. All procedures are performed carefully to minimize the creation of splashes or aerosols. 7. Work surfaces are decontaminated on completion of work or at the end of the day and after any spill or splash
Laboratory Biosafety Level Criteria-Biosafety Level 2 of viable material with disinfectants that are effective against the agents of concern res.stocks,and othe regulated wastes are de- tamination method such as autoclavng.Materials to be decontaminated outside of the imme diate labo placseeonanrabeeaeakpe taminated off-site from the facility are packaged in a ccor dance with appicable bca hd federal regu panat0dogsntcantopogranieneoeeg B.Special Practices 1. Access to the laboratory is limited or restricted by the laboratory director when work with infectious agents is in me. serious consequences,are not allowed in the laboratory or animal rooms mple,persons who ar be at cquiring infections. ineiaboao director has the sing eac the labora tory or anim al room work in 2 The labe bi the potental hazards and meet specific entry require- ments (e.g.,im mu nization)may enter the labor atory A biohazard sign must be posted on the entrance to the sare in biosafety level.the requred investigator's name and telephone number,any personal 22
Laboratory Biosafety Level Criteria – Biosafety Level 2 22 of viable material with disinfectants that are effective against the agents of concern. 8. All cultures, stocks, and other regulated wastes are decontaminated before disposal by an approved decontamination method such as autoclaving. Materials to be decontam inated outside of the imme diate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Materials to be decontaminated off-site from the facility are packaged in accordance with applicable local, state, and federal regulations , befo re rem oval from the fa cility. 9. An insect and rodent control program is in effect (see Appendix G). B. Special Practices 1. Access to the laboratory is limited or restricted by the labor atory d irecto r whe n work with infe ctiou s age nts is in progress. In general, persons who are at increased risk of acquiring infection, or for whom infection may have serious consequen ces, are not allowed in the laboratory or animal room s. For examp le, persons who are immunocomprom ised or immunosuppressed m ay be at increased risk of acquiring infections. The laboratory director has the final responsibility for assessing each circu ms tanc e and determining w ho m ay enter or w ork in the labora tory or anim al room . 2. The laboratory director establishes policies and procedures whereby only persons who have been advised of the potential hazards and meet specific entry requireme nts (e .g., im mu nizatio n) m ay enter the labor atory. 3. A biohazard sign must be posted on the entrance to the laborator y when etiolo gic agen ts are in us e. Appro priate information to be posted includes the agent(s) in use, the biosafety level, the required immunizations, the investigator’s name and telephone number, any personal
ratory BiosafetyLevel Criteria -Biosafety Level 2 o ratory. the labora tory (e.g.hepatitis B vaccine or TB skin test- ing). 5 When appropriate.considering the agent(s)handled baseline serum samples for laboratory and other at-risk lly de 6. Biosafety proce rated inte ct operatin ng proceduresorna biosafety manual adopted or prepare the lab atory by the laborator are required to read and follow instructions on practices and procedures. The laboratory director ensures that laboratory and support personnel receive pot ning on the the exposure evaluationp res.Personnel receive training as necessary fo A high degree of precaution mustal ways be n with ringes.slides.pipettes.capillary tubes.and scalpels. when theres no altemative.such as parentera lastic ware should be substituted for glassware whenever possible 23
Laboratory Biosafety Level Criteria – Biosafety Level 2 23 prote ctive e quipm ent th at m ust be worn in th e labo rator y, and an y proced ures req uired for e xiting the labo ratory. 4. Laboratory personnel receive appropriate immunizations or tes ts for the agents handled or po tentia lly pres ent in the labora tory (e.g., hep atitis B vacc ine or TB skin tes ting). 5. When appropriate, considering the agent(s) handled, baseline serum samples for laboratory and other at-risk personnel are collected and stored. Additional serum specimens may be collected periodically, depending on the agents handled or the func tion of the facility. 6. Biosafety procedures are incorporated into standard operating procedures or in a biosafety manual adopted or prepared specifically for the laboratory by the laboratory director. Personnel are advised of special hazards and are required to read and follow instructions on practices and procedures. 7. The laboratory director ensures that laboratory and support personnel receive appropriate training on the potential hazards associated with the work involved, the necessary precautions to prevent exposures, and the exposure evaluation procedu res. Personnel receive annual updates or additional training as necessary for procedural or policy changes. 8. A high de gree of p recaution mus t always be taken w ith any co ntam inated sha rp item s, inc luding need les an d syringes, slide s, pipettes , capillary tubes , and sc alpels. a. Need les and s yringes or o ther sha rp instrum ents should be restricted in the la bora tory fo r use only when there is no alternative, such as parenteral injection, phlebotomy, or aspiration of fluids from laboratory animals and diaphragm bottles. Plasticware should be substituted for glassware whenever possible
Laboratory Biosafety Level Criteria-Biosafety Level 2 b.Only needle-locking sheared.broken.recapped.removed from dispos- able syringes,or otherwise manipulated by hand used for sharps disposal.Non-disposable sharps must be ntain autoclaving c. appropriate. d.Broken glassware must not be handled directly by hand,but must be removed by mechanical means 5,0 ment,and broken glass are deconta minated efore ogbanngcordingoanylocalstate,ortederal Cultures,tissues,speci en of body fluids,or potentially that prevents leakage during collection.handling. proces sing.storage,trans port,or shipping. 10. l aboratoryeauinment and work surfaces should be de effecive disinfectant on a routine .splashest0r8hernoigrnand tion by infectious materials.Contaminated equipment it is s nance or packaged for trans port in aco ord ance applic ore lity. 24
Laboratory Biosafety Level Criteria – Biosafety Level 2 24 b. Only needle-locking syringes or disposable syringeneedle units (i.e., needle is integral to the syringe) are used for injection or aspiration of infectious materials. Used disposa ble need les mu st not be b ent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal; rather, they must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving. c. Syringes which re-sheathe the needle, needleless systems, and other safety devices are used when appropriate. d. Broken glassware must not be handled directly by hand, but must be removed by mechanical means such a s a brus h and du stpan, ton gs, or forc eps. Containers of contaminated needles, sharp equipment, and brok en glass are decontam inated before disposal, according to any local, state, or federal regulations. 9. Cultu res, tis sues, sp ecim ens of bo dy fluid s, or p oten tially infectious wastes are placed in a container with a cover that prevents leakage during collection, handling, proces sing, stora ge, trans port, or sh ipping. 10. Laboratory equipment and work surfaces should be decontaminated with an effective disinfectant on a routine basis, after work with infectious materials is finished, and especially after overt spills, splashes, or other contamination by infectious materials. Contaminated equipment must be decontaminated according to any local, state, or federal regulations before it is sent for repair or maintenance or pack aged fo r transpo rt in accord ance w ith applicable local, state, or federal regulations, before rem oval from the fa cility
Laboratory Biosafety Level Criteria -Biosafety Level 2 11. vert expc dthe labo ratory director.Medical evaluation,survellance,and treatment are vid das approprate and wrtten re 12. the or bngoed arenot C.Safety Equipment(Primary Barrers) 1. ets.preferably Procedures with a potential fo shakingormixing. uption open ing contan animals intranasally,and harvesting nfected tissues from animals or embryonate eggs. b.High concentrations or large volumes of infectious ma may b fuge safety cups are used,and if these rotors or 2. Face protection(goggles.mask.face shield orother microorgmmstbemaniputated outsd 3 25
Laboratory Biosafety Level Criteria – Biosafety Level 2 25 11. Spills and a ccidents that result in o vert expo sures to infectious materials are immediately reported to the laboratory director. Medical evaluation, surveillance, and treatment are provided as appropriate and written records are maintained. 12. Animals not involved in the work being performed are not perm itted in the lab. C. Safety Equipment (Primary Barriers) 1. Prop erly m aintained b iologic al saf ety cabinet s, pre ferably Class II, or other appropriate personal protective equipment or physical containment devices are used whenever: a. Procedures with a potential for creating infectious aerosols or splashes are conducted. These may include centrifuging, grinding, blending, vigorous shaking or mixing, sonic disruption, opening containers of infectious materials whose internal pressures may be different from ambient pressures, inoculating animals intranasally, and harvesting infected tissues from animals or embryonate eggs. b. High concentrations or large volumes of infectious agents are used. Such materials may be centrifuged in the open labor atory if seale d roto r hea ds or centrifuge safety cups are used, and if these rotors or safety cu ps are o pened only in a biologica l safety cabinet. 2. Face protection (goggles, mask, face shield or other splatter gu ard) is use d for antic ipated spla shes or s prays of infectious or other hazardous materials to the face when the microorganisms must be manipulated outside the BSC. 3. Protective laboratory coats, gowns, smock s, or uniforms designa ted for lab u se are w orn while in th e laborato ry