Institutional BIOSAFETY MANUAL

Transcription

1 Institutinal BIOSAFETY MANUAL The purpse f this manual is t define the bilgical safety plicies and prcedures pertaining t research peratins at Bstn University and Bstn Medical Center. These plicies and prcedures are designed t safeguard persnnel and the envirnment frm bilgically hazardus materials and t cmply with federal, state, and lcal regulatry requirements. All BU and BMC Principal Investigatrs and labratry wrkers must adhere t the bilgical safety plicies and prcedures in the cnduct f their research and the management f their labratries. Fr infrmatin abut specific bilgical safety prgrams fr peratins nt cvered in this manual, cntact the Institutinal Bisafety Cmmittee ffice r the Bisafety Officer. Chair, Institutinal Bisafety Cmmittee Assciate Vice President, Research Cmpliance Page 1 f 201

2 Institutinal BIOSAFETY MANUAL TABLE OF CONTENTS Page Chapter 1 Bilgical Safety Prgram: Purpse, Scpe, and Respnsibilities 4 Chapter 2 Apprval f Research Prjects 12 Chapter 3 List f Regulatins and Guidelines 20 Chapter 4 Bisafety Principles 22 Chapter 5 Labratry Bisafety Practices 30 Chapter 6 Labratry Training 43 Chapter 7 Decntaminatin and Sterilizatin 47 Chapter 8 Bihazardus Spill Respnse 52 Chapter 9 Bihazardus and Medical Waste Dispsal 58 Chapter 10 CDC/USDA Select Agents 61 Chapter 11 Transprtatin f Bilgical Materials 78 Page 2 f 201

3 Appendix A IBC Applicatin Frms/Bilgical Use Authrizatin (BUA) 92 Appendix B Imprtatin and Exprtatin f Etilgic Agents 93 Appendix C Labratry Ventilatin and Cntainment fr Bisafety 96 Appendix D Autclave Quality Assurance Prgram 102 Appendix E Bisafety Level 2 (BSL-2) Requirements 105 Appendix F Bisafety Level 3 (BSL-3) Facility Standard Operating Prcedure (SOP) 110 Appendix G Bisafety Level 4 (BSL-4) Requirements 127 Appendix H Guidelines fr Wrk with Txins f Bilgical Origin 136 Appendix I List f Bilgical Agents with the Ptential t Cause Labratry 140 Acquired Infectin (LAI) in Use at Bstn University Appendix J Prin Research/Cruetzfeldt-Jacb Disease (CJD) Guidelines 141 Appendix K Summary f Requirements fr Bisafety Levels 144 Appendix L Summary f Requirements fr Animal Bisafety Levels 146 Appendix M Bldbrne Pathgen Standard 148 Appendix N Wrking Safely with Animals 155 Appendix O Prcedures fr Wrking in an Animal Bisafety Level (ABSL-2) Facility at BU Appendix P Plicy fr Verifying the Identity f Attenuated Pathgens 163 Appendix Q Bstn Public Health Cmmissin Requirements 165 Appendix R ROHP Medical Surveillance Prgram 166 Appendix S Labratry and Equipment Decntaminatin Prcedures 174 Appendix T Labratry Dr Signage 183 Appendix U Bstn University and Bstn Medical Center Research 184 Cmpliance Plicy n Disease Surveillance and Reprting fr High-Risk Agents Appendix V Institutinal Bisafety Cmmittee Oversight Prgram 191 Appendix W Criteria fr Develpment f Standard Operating Prcedures (SOP) 195 Page 3 f 201

4 Chapter 1 Bilgical Safety Prgram: Purpse, Scpe, and Respnsibilities Purpse The purpse f this (BSM) is t define plicies and prcedures pertaining t use f bilgical materials in research at Bstn University (BU) and Bstn Medical Center (BMC). These plicies and prcedures are designed t safeguard persnnel and the envirnment frm bilgically hazardus materials withut unduly limiting academic research. This manual als ffers guidelines t cmply with federal and state regulatry requirements. The wrk practices, prcedures, and plicies specified in this manual are based n current regulatry requirements and accepted best bisafety practices. Implementatin f these measures will reduce the likelihd that an incident invlving a bilgical agent will ccur and will fulfill regulatry bisafety expectatins. Labratry micrbilgical wrk usually invlves ptential expsure t bilgical hazards, as well as t chemical and radilgical hazards. Cnsequently, this manual shuld be used in cnjunctin with the BU Chemical Hygiene Plans and Radiatin Safety Manual, respectively. Fr infrmatin abut specific bilgical safety prgrams fr peratins nt cvered in this manual, cntact the Institutinal Bisafety Cmmittee (IBC) ffice r the Bisafety Officer (BSO). Scpe This manual applies t all BU and BMC research activities invlving bilgical agents. All faculty, staff, students, and visitrs wh wrk n BU- r BMC-spnsred prjects r at BU r BMC facilities are included in the scpe f this manual. Bilgical agents include all infectius micrrganisms (bacteria, fungi, parasites, prins, rickettsias, viruses, etc.) that can cause disease in humans r pse significant envirnmental r agricultural impact, as well as the txins derived frm such rganisms. Additinally, recmbinant DNA; human r nn-human primate tissues, fluids, cells, r cell cultures; transgenic plants r animals; and any wrk with animals and their tissues, which are knwn t be reservirs f zntic diseases, are whlly r partly cvered by the prcedures and plicies in this manual. Nte: In this dcument, the term Institutin is used t refer t the cmbined BU and BMC entities. Bilgical Safety Prgram Gals The gals f the Bilgical Safety Prgram, referenced in this manual as the Bisafety Prgram, are t prtect labratry wrkers, the public, and the envirnment frm ptentially hazardus bilgical agents. The IBC advcates the use f bisafety precautins that effectively reduce r eliminate the risk f expsure t ptentially hazardus agents used in research. In develping its guidelines, the IBC is ensuring that all plicies and prcedures are in accrdance with bth the regulatry framewrks gverning the use f bilgical materials and the best practices adpted natinally. Page 4 f 201

5 Chapter 3 cntains a listing and summary f the regulatins and guidelines that gvern the use f bilgical materials in research. Rles and Respnsibilities Success f the Bisafety Prgram, like any ther safety prgram, requires a team effrt invlving the IBC, Principal Investigatrs, labratry wrkers, the Research Occupatinal Health Prgram (ROHP), and Envirnmental Health and Safety (EHS). Principal Investigatrs are respnsible fr the health and safety f persnnel wh wrk under their supervisin and ccupy their labratry space. BU and BMC administratin, the IBC, and EHS endrse this manual and encurage active participatin in maintaining high standards at BU and BMC. Assciate Vice President, Research Cmpliance (AVP-RC) The AVP-RC has verall respnsibility fr: Oversight fr the cntrl f hazards in the research labratries and fr ensuring that cmprehensive, enterprisewide prgrams are in place fr the safe handling f all hazardus (e.g., bilgical, chemical, radilgical, etc.) materials. All nn-financial research cmpliance at BU and BMC. Direct functinal respnsibility fr the IBC, Bisafety Prgram, EHS, labratry safety cmmittees, bisafety prgrams, labratry animal use and care prgrams (bth Institutinal Animal Care and Use Cmmittees (IACUC) and the animal care prgrams), respnsible cnduct f research, spnsred prgrams, and ther research-related versight cmmittees. Acts as the Respnsible Official (RO) fr the City f Bstn s Public Health Cmmissin labratry regulatins. Develps and ensures cmmunicatin between IBC, IRB, IACUC, spnsred research ffices, and regulatry agencies (e.g., City f Bstn, CDC). In cnsultatin with prvsts and deans at bth the medical campus and the Charles River campus, as well as the BMC leadership, appints varius cmmittee members. The IBC, the Bisafety Prgram Directr, the Research Occupatinal Health Prgram, and EHS have been charged with planning and implementing the Bisafety Prgram, the purpse f which is t ensure the health and safety f all persnnel wrking with bihazardus r infectius agents and recmbinant DNA. Institutinal Bisafety Cmmittee The Institutinal Bisafety Cmmittee (IBC) is respnsible fr verall versight f the Bisafety Prgram at BU and BMC (See Appendix V fr IBC versight Prgram). The IBC carries ut these functins pursuant t requirements set frth by the Natinal Institutes f Health (NIH), the Centers fr Disease Cntrl and Preventin (CDC), Occupatinal Safety and Health Administratin (OSHA), the City f Bstn Public Health Cmmissin (BPHC), the Massachusetts Department f Public Health (DPH), and Bstn University. Page 5 f 201

6 The IBC s respnsibilities include: Overall versight f the Institutinal Bisafety Prgram at BU and BMC, including develpment f new, and review f existing, plicies and prcedures designed t enhance the bilgical safety prgrams. Reviews and apprves training prgrams and establishes qualificatins fr individuals wrking with bilgical materials. Reviews and apprves gene transfer clinical trials. Crdinates the bilgical safety requirements with ther campus-wide cmmittees (e.g., IACUC) r prgrams (e.g., Research Occupatinal Health Prgram). Reviews and apprves new research prpsals invlving rdna and bihazardus material in accrdance with guidelines established by the BPHC, DPH, OSHA, U.S. Department f Agriculture (USDA), CDC, NIH, BU and BMC, as well as maintains prject apprval and reviews amendments. Sets required cntainment levels fr research prjects. Generally, bisafety levels (BSL) established by the CDC and NIH will be used as the level f cntainment; hwever, the IBC has the authrity t increase r decrease the level f cntainment accrding t the prject s specific circumstances. Hwever, the IBC als has the authrity t upgrade labratry cntainment levels if the prtcl review identifies specific hazards assciated with the prpsed peratins. Apprves design specificatins and criteria fr cntainment facilities develped by the Bisafety Officer and cnducts the final review and acceptance f BSL-3 and BSL-4 labratries. Develps cmprehensive inspectin prgrams, as well as receives and reviews the findings f such inspectins. Investigates vilatins f bisafety prcedures r plicies and significant accidents r illnesses invlving bilgical agents. If apprpriate, recmmends disciplinary actin t the prper BU/BMC fficials. Reviews and apprves annual reprts sent t: Bstn Public Health Cmmissin, under the rdna rdinance NIH, cncerning rdna expsures incidents NIH s Office f Bitechnlgy Activities, an IBC update Reviews the SA Bisafety Plan Amends the List f Bilgical Agents with the Ptential t Cause Labratry Acquired Infectin (LAI) in use at Bstn University (see Appendix I), as needed, based n the review f prpsed research invlving bihazardus materials; Ensures PIs and research staff listed n apprved IBC prtcls with these agents (in Appendix I) receive agent specific ID cards and Agent Infrmatin Sheets (AIS), in crdinatin with ROHP. Other activities as delegated by the Prvst r the AVP-RC Page 6 f 201

7 Bisafety Officer The Bisafety Officer (BSO) is respnsible fr prviding guidance n the safe handling f bilgical agents and verall management f the Bisafety Prgram. The BSO is an ex-ffici member f the IBC. The BSO s specific respnsibilities include, but are nt necessarily limited t, the fllwing: Prvides technical advice t the IBC, and researchers n labratry cntainment, security, and safety prcedures. Oversees peridic and unscheduled inspectins t ensure that labratry standards are rigrusly maintained. Develps emergency plans fr handling spills and persnnel cntaminatin. Develps and implements labratry safety practices. Develps design specificatins and criteria fr cntainment facilities. Crdinates the preparatin f IBC reprts submitted t regulatry agencies. Prvides training prgrams as apprved by the IBC. The BSO regularly reprts n the Bisafety Prgram t the IBC. The BSO s reprt shuld include rutine peratinal updates and any significant prblems r vilatins f the regulatry mandates r IBC requirements n any research-related accidents r illnesses that have ccurred. Research Safety Directr The Research Safety Directr (RSD) has verall respnsibility and wrks with the different safety cmmittees t ensure that labratry research is cnducted in apprpriate facilities, using safety equipment, and implementing safety prcesses. The RSD is als: Appinted as the Respnsible Official under the CDC Select Agents regulatin Crdinate wrk with apprpriate regulatry agencies and acts as a liaisn during regulatry inspectins. Wrks with the AVP-RC, IBC, and ther institutin fficials t crdinate respnses t any regulatry findings. Review new and prpsed regulatins and ther requirements and summarize impact t the institutin. Determine prgrams and prcesses necessary fr an effectively safe prcess t cnduct labratry research. Page 7 f 201

8 Envirnmental Health and Safety The varius prgrams within Envirnmental Health and Safety (EHS) wrk clsely with the BSO t ensure that peratins fr which EHS has respnsibility are cnducted in accrdance with the criteria and guidelines established by the BSO. These include: Dispsal f medical waste. Selectin f apprpriate prtective clthing fr individuals wrking with hazards, including bihazardus, materials. Develpment and implementatin f emergency respnse and preparedness plans. Mnitring wrk areas, including the presence f allergens. Principal Investigatrs Principal Investigatrs (PIs) r Labratry Directr are respnsible fr the health and safety f all persnnel and cmpliance with all applicable regulatins and the criteria established in this manual in their labratries. The PI: Ensures that specific labratry hazards are effectively cmmunicated t labratry persnnel; persnnel have received apprpriate training and are cmpetent t perfrm prcedures used in the labratry; and cntrls are in place t minimize risks assciated with these hazards. Develps labratry-specific standard perating prcedures (SOPs) that cver the hazards and activities (bth rutine activities and unusual events) relevant t the labratry. Ensures that engineering cntrls are available, in gd wrking rder, and are used apprpriately t minimize expsure t bihazardus agents. Ensures that apprpriate persnal prtective equipment is available and used by labratry persnnel. Ensures that all labratry persnnel receive general bisafety training that is cnducted as part f the Bisafety Prgram, as well as specific training n the hazards, prcedures, and practices relevant t the labratry in which they are wrking. All training must be dcumented and recrds maintained. Ntifies the IBC and btains prir IBC apprval fr wrk invlving recmbinant DNA and/r bihazardus material and cnfrms t all terms and cnditins f IBC apprval. Ensures that labratry wrkers are prvided immunizatins and medical surveillance prir t, and in the event f, expsure t bihazardus agents as apprpriate (based n current CDC and IBC recmmendatins). Immunizatins are available thrugh BU s Research Occupatinal Health Prgram (617) , fr emplyees. Page 8 f 201

9 Ntifies the BUMC/BMC Cntrl Center at (617) r EHS emergency telephne, (617) at the Charles River Campus, f any spills r incidents invlving bilgical agents that result in expsure t labratry persnnel r the public, r release t the envirnment. The Cntrl Center will ntify the BSO. Ensures that bilgical agents are dispsed f accrding t regulatins, as utlined in this manual. Ensures that bihazardus materials t be transprted are packaged and shipped in accrdance with regulatins. Ensures that peridic inspectins f the labratry are cnducted. Labratry Safety Crdinatrs Supprts the PI and versees that safety practices are implemented in the lab s daily peratins. Authrized t represent the PI in matters related t the implementatin f labratry and wrker safety. Serve as the primary labratry cntact with EHS fr issues related t safety (i.e. bilgical, chemical, fire, general safety, cntrlled substances, etc). Take psitive actins t help reduce the ptential fr accidents and incidents assciated with labratry peratins. Infrm labratry persnnel and/r students f the safety hazards assciated with their wrk. Instruct all labratry persnnel and students in safe wrk methds. Reprt all accidents, near misses, r safety cncerns t the PI and EHS. Ensure that apprpriate Standard Operating Prcedures (SOP) are established and that lab persnnel and students are apprpriately trained and fllw them. Wrk with EHS t determine best safe practices and prcedures. Wrk with EHS t ensure that lab persnnel and students cmplete all required safety trainings in a timely manner. Ensure that all deficiencies identified by EHS r utside regulatry inspectrs are addressed and crrected within the time required. Participate in the incident review prcess. Stp peratins that are in clear vilatin f the safety requirements, apprved SOPs, r may ptentially result in injuries r ptential expsures. Research Occupatinal Health Prgram (ROHP) and Occupatinal Health Officer (OHO) The Occupatinal Health Officer (OHO) is the leader f the Research Occupatinal Health Prgram (ROHP) and is respnsible fr reprting f BU/BMC expsure incidents invlving select agents (see Chapter 10 CDC/USDC Select Agents) and return t wrk plans t the Bstn Public Health Cmmissin (BPHC). ROHP is primarily respnsible fr establishing and perfrming apprpriate medical surveillance fr all persnnel perfrming research r supprting research such as animal care wrkers, Page 9 f 201

10 facilities, EHS, Plice and Public Safety. Surveillance is required at the time f hire r transfer int the research envirnment and peridically depending n the wrk envirnment, ccupatinal expsure and risk fr each psitin r jb categry. In additin t perfrming medical surveillance, ROHP is als respnsible fr: Develp expsure treatment plans fr labratry expsures r incidents, including pst-expsure management and mnitring Review IBC prtcl prpsals and determine additinal medical surveillance requirements fr PI s and ther persnnel included in prpsals befre apprvals are granted; create and issue wallet-sized agent cards, agent infrmatin sheets, and emergency medical respnse prtcls t thse persnnel apprved by the IBC t wrk with bilgical agents with the ptential t cause LAI (see Appendix I, List f Bilgical Agents with the Ptential t Cause LAI in Use at Bstn University) Participates in labratry safety, IACUC, radiatin safety and ther cmmittee meetings as required t ensure apprpriate health and safety practices are fllwed Assists PI s and EHS in the preparatin and presentatin f bisafety and agent specific training Prvide medical supprt cverage n a 24/7 basis fr researchers and ther persnnel t call fr triage, evaluatin, and medical care referral based n severity, lcatin and time f labratry expsure r incident Wrk with EHS t develp SOPs and apprpriate health and safety practices Perfrm return t wrk assessments in cnjunctin with BPHC fr labratry expsures invlving high risk agents Issue generic wallet-sized agent cards t persnnel with ptential expsure t hazardus materials in research labratries and animal care facilities Labratry Wrkers Labratry wrkers are the mst imprtant element in develping and maintaining a safe labratry envirnment. Labratry wrkers are respnsible fr their wn health and safety, as well as that f their cwrkers. An incident caused by ne labratry wrker can have a widespread effect n thers. Labratry wrkers are expected t: Fllw prcedures and practices established by BU r BMC, the IBC, and the labratry. Use best bisafety labratry practices t minimize expsures t bilgical agents and t avid ther incidents (such as persnal injuries, chemical and radiatin spills, labratry fires, explsin, etc.). Attend the required Labratry Safety Training initially and cmplete the annual refresher training thereafter. Page 10 f 201

11 Reprt spills, accidents, and unsafe labratry cnditins t the PI, EHS, and/r ther respnsible parties. Utilize cntrl measures, such as bilgical safety cabinets and persnal prtective equipment, t prevent expsure t bilgical agents and cntaminatin f persnnel and facilities. Immediately cntact ROHP at (617) (ROHP) in the event f an expsure injury where medical triage and evaluatin, dcumentatin and ntificatins can be perfrmed. Page 11 f 201

12 Chapter 2 Apprval f Research Prjects Wh Needs Apprval Principal Investigatrs (PIs) at BU and BMC planning t carry ut research using recmbinant DNA and/r bilgically hazardus materials that pse a ptential risk t the health f humans r animals, either directly thrugh infectin r indirectly thrugh damage t the envirnment, shuld submit an IBC Applicatin entitled Bilgical Use Authrizatin (BUA) t the IBC ffice fr review and apprval by the IBC. Nte: Because the Bstn Public Health Cmmissin (BPHC) requires registratin f BSL-1 rdna prjects, studies designated Bisafety Level 1 (BSL-1) and cnsidered "exempt" by the NIH must be reviewed and apprved by the IBC. Researchers using any f the fllwing shuld cmplete and submit a Bilgical Use Authrizatin fr review and apprval: Recmbinant DNA Infectius agents Txins Human bld, bdy fluids, unfixed tissue, r human cell cultures Tissues, rgan, r cell lines cultures f human r nn-human primate rigin Creatin r certain uses f transgenic animals Human gene therapy Nn-human primates (because f the ptential t transmit herpes and ther diseases) Sheep and any tissues derived frm them (these tissues can transmit Cxiella burnetii, the causative agent f Q-fever) Transgenic plants Wild animals knwn t carry zntic diseases (e.g., bats, raccns, etc.) When wrking with ptentially infectius agents and human subjects r experimental animals, IBC review is necessary in additin t review by the Institutinal Animal Care and Use Cmmittee (IACUC) r the apprpriate Institutinal Review Bard (IRB). Page 12 f 201

13 PIs whse research will invlve bihazardus materials, infectius agents, human tissue (e.g., bld, cells, fluids, etc.) r whse research cmes under the gvernance f any f the fllwing campus rules r gvernmental regulatins, are required t cmplete a Bilgical Use Authrizatin and btain IBC apprval prir t receiving such materials r cmmencing such research. PIs are als required, where applicable, t maintain a medical surveillance prgram fr labratry emplyees (see Appendix O). Relevant campus and/r gvernmental regulatins include the fllwing: NIH Guidelines fr Research Invlving Recmbinant DNA Mlecules Bstn Public Health Cmmissin OSHA Bldbrne Pathgens Standard OSHA Tuberculsis Standard Massachusetts Department f Public Health The Centers fr Disease Cntrl and Preventin (CDC) Select Agent regulatins Massachusetts Department f Health and Human Resurces Medical Waste Management Act Internatinal, federal, and state transprt regulatins Nte: If a Principal Investigatr is perfrming this type f wrk withut IBC apprval, he r she is ut f cmpliance with current NIH and lcal regulatins and has placed the institutin in that psitin as well. New Applicatins A new Bilgical Use Authrizatin must be submitted and reviewed by the IBC fr any research using rdna and/r bilgically hazardus materials. PIs seeking IBC apprval fr the first time als need t submit a curriculum vitae (CV) with their applicatin. One applicatin fr bth rdna and bihazardus wrk may be submitted via the Research Infrmatin Management System (RIMS), and this nline system may be accessed frm the IBC website, IBC apprval f recmbinant DNA and bihazardus research prjects is effective fr three years. PIs must cmplete a renewal frm annually t cntinue wrk fr up t three years after the initial apprval. After three years, the applicatin must be resubmitted and reviewed by the cmmittee. Renewals A renewal ntice serves as a mechanism fr the PI t prvide an annual update and this frm is sent t the PI listed n the riginal apprval the first and secnd year after initial apprval f a prtcl. The PI is asked t list all prpsed deviatins frm the prtcl as initially apprved (r since the last renewal ntice); changes in labratry lcatin; changes in labratry staff wrking n the prject; and any prject titles t be added. Page 13 f 201

14 If there are significant deviatins frm the prtcl, especially deviatins that affect the cntainment level (i.e., new study rganisms, a new hst-vectr-dnr system, r any ther mdificatins that may affect the cntainment level), the IBC may ask the PI t seek an additinal apprval t cver the additinal experiments. When a prject is renewed as part f the annual update prcess, all new lab staff must submit either an Initial Health Questinnaire r an Annual Health Questinnaire (Initial, if n prir baseline medical histry n recrd) t ROHP and cmplete lab safety training. Fr changes in PI, the new PI must attach his r her CV (tw-page NIH frmat) t the renewal frm. Amendments Amendments must be submitted in electrnic r hard cpy frm fr changes within an apprved prject. All changes shuld be detailed in the Prtcl Amendment frm, which the IBC must review and apprve. Title additins (expedited review) apprval may be applied t several different granting agencies, but all grant titles must be registered with the IBC. Lab space additins (expedited review) apprval applies nly t wrk perfrmed in registered lab space. Fr nn-pi persnnel changes (expedited review), individuals must be trained in lab techniques and have cmplied with necessary trainings r apprval prcedures, such as medical surveillance and lab safety training. If technical changes (full cmmittee review) are extensive, the IBC may require the PI t submit a cmpletely new applicatin. A change in PI als requires full cmmittee review. The new PI must attach his r her CV (tw-page NIH frmat) t the amendment. Bihazardus and Ptentially Infectius Materials Categries Bihazards are infectius agents r bilgically derived infectius materials that present a risk r ptential risk t the health f humans r animals, either directly thrugh infectin r indirectly thrugh damage t the envirnment. Infectius agents have the ability t replicate and give rise t ptentially large ppulatins in nature when small numbers are released frm a cntrlled situatin. The fllwing is a listing f the ptentially hazardus bilgical materials and agents. PIs shuld fllw the instructins in the Bilgical Use Authrizatin applicatin carefully t ensure that all apprpriate sectins f the applicatin are cmpleted. If a PI intends t use bilgical agents that are nt listed in this sectin, he r she shuld cntact the IBC r BSO fr advice regarding prper cmpletin f the Bilgical Use Authrizatin. Human, animal, and plant pathgens Viruses, including ncgenic and defective viruses Rickettsiae Chlamydiae Bacteria, including thse with drug-resistant plasmids Fungi Parasites Undefined r ther infectius agents, such as prins Page 14 f 201

15 All human bld, bld prducts, tissues, and certain bdy fluids Cultured cells (all human r certain animal, including nn-human primates) and the ptentially infectius agents these cells may cntain Allergens Txins (bacterial, fungal, plant, etc.) Certain recmbinant nucleic acid prducts Clinical and diagnstic specimens Infected animals and animal tissues Nn-human primates and any tissues derived frm them (can transmit Herpes B virus) Sheep and any tissues derived frm them (can transmit Cxiella burnetii, the causative agent f Q- fever) Nte: The IBC has develped a Plicy fr Verifying the Identity f Attenuated Pathgens (Appendix P) that must be fllwed by all researchers. Recmbinant DNA (rdna) Materials Generatin r Use f rdna The NIH s Guidelines fr Research Invlving Recmbinant DNA Mlecules is the definitive regulatry reference fr recmbinant DNA (rdna) research in the United States. There may be experiments, nt cvered by the guidelines that wuld require review and apprval by utside agencies befre initiatin. If the experimental prtcl is nt cvered by the NIH s guidelines, cntact the BSO at (617) at BUMC and (617) at CRC t determine further review requirements. Nte: rdna wrk with BSL-4 classificatin is nt permitted in Bstn. Human Gene Therapy All prtcls invlving human gene therapy must be submitted t the NIH Office f Bitechnlgy Activities t underg initial review by the Recmbinant Advisry Cmmittee (RAC). After RAC review, the prtcl must be apprved by the IBC and IRB. Fr mre details abut IBC apprval f human gene therapy prtcls, call (617) Fr infrmatin abut IRB submissins, call (617) (BUMC) r (617) (CRC). Use f Animals The use f animals in research requires cmpliance with the Animal Welfare Act, administered by the USDA s Animal and Plant Health Inspectin Service (APHIS); the Public Health Service Plicy n Humane Care and Use f Labratry Animals, administered by NIH s Office f Labratry Animal Welfare (OLAW); and all applicable state r lcal regulatins cvering the care and use f animals. All prtcls invlving the use f live animals must be reviewed and apprved by the IACUC befre their implementatin. All PIs planning t use rdna and/r bihazardus materials in their labratry r in research animals must receive IBC apprval. Page 15 f 201

16 Transgenic Animals PIs wh create transgenic animals, either in the PI s lab r thrugh the BU Transgenic Cre facility, as well as PIs wh use transgenic animals at ABSL-2, r at ABSL-1 if nt cnsidered exempt under the NIH Guidelines (Sectin III-E, Appendix C-VIII) must cmplete an IBC applicatin and submit it t the IBC fr apprval prir t initiatin f experimentatin. In additin, the IACUC must apprve the prtcl. The IACUC can be reached at (617) (BUMC) r (617) (CRC). Tissue Culture/Cell Lines Risk Grup 1/Bisafety Level 1 (BSL-1) The fllwing are cnsidered Risk Grup 1 cell lines and are handled at BSL-1: They are nn-primate rigin They d nt harbr a primate virus They are nt cntaminated with bacteria, mycplasma, r fungi They are well established after several years f use in the labratry nt t cause infectin in humans. Nte: All human and nn-human primate cell lines must be used at BSL-2 as they may harbr previusly undefined pathgens. Risk Grup 2/Bisafety Level 2 (BSL-2) When cell cultures are knwn t cntain an etilgic agent r an ncgenic virus, the cell line can be classified at the same level as that recmmended fr the agent r virus. The CDC has recmmended that all cell lines f human rigin be handled at BSL-2. The fllwing are identified as Risk Grup 2 and must be handled at BSL-2: Primate cell lines derived frm lymphid r tumr tissue All cell lines expsed t r transfrmed by a primate ncgenic virus All clinical material (e.g., samples f human tissues and fluids btained after surgical resectin r autpsy fr use in rgan culture r establishment f primary cell cultures) All primate tissue All cell lines new t the labratry (until prven t be free f all adventitius agents) All virus and mycplasma-cntaining primate cell lines Risk Grup 3/Bisafety Level 3 (BSL-3) When cell cultures are knwn t cntain any Risk Grup Three bilgical agent, the cell line can be classified at the same level as that recmmended fr the agent r virus. The fllwing are examples f bilgical materials identified as Risk Grup 3 and must be handled at BSL-3: High titer HIV wrk St. Luis encephalitis virus Venezuelan equine encephalmyelitis virus Page 16 f 201

17 M. tuberculsis Cncentrated Lentivirus r Lentiviral vectrs with high likelihd f aersl frmatin. Francisella tularensis See Appendix-F: BSL-3 Facility Standard Operating Prcedures. Risk Grup 4/Bisafety Level 4(BSL-4) When cell cultures are knwn t cntain any Risk Grup Fur bilgical agent as listed in BMBL (Fifth editin), the cell line can be classified at the same level as that recmmended fr the agent r virus. The fllwing are examples f bilgical materials identified as Risk Grup 4 and must be handled at BSL-4: Marburg virus Ebla virus Junin virus Machup virus Guanarit virus See Appendix-G: BSL-4 Requirements. Nte: Appendix K prvides a summary f Bisafety Level requirements fr each risk grup. Tuberculsis Since 1985, the incidence f tuberculsis in the United States has been increasing steadily, reversing a 30- year dwnward trend. Recently, drug-resistant strains f Mycbacterium tuberculsis have becme a serius cncern. Outbreaks f tuberculsis, including drug-resistant strains, have ccurred in health-care envirnments. In 2005 the CDC published Guidelines fr Preventing the Transmissin f Mycbacterium tuberculsis in Health-care Settings. MMWR 2005; 54 (N. rr-17, 1-141). (Errata published September 25, 2006.) The guidelines cntain specific infrmatin n ventilatin requirements, respiratry prtectin, medical surveillance, and training fr thse persnnel wh are cnsidered at-risk fr expsure t tuberculsis. Fr mre infrmatin, cntact the BSO. PIs intending t wrk with Mycbacterium tuberculsis in the labratry must btain written apprval frm the IBC via the IBC applicatin prcess befre beginning wrk. Prpagatin and manipulatin f Mycbacterium tuberculsis cultures must be perfrmed at Bisafety Level 3 (BSL-3). PI s and all ther persns apprved t wrk with M. tuberculsis are required t underg bi-annual tuberculsis screenings. Page 17 f 201

18 Vaccinia Virus PIs wishing t use vaccinia virus must btain written apprval frm the IBC via the applicatin prcess. BSL-2 practices and prcedures must be fllwed. All emplyees wh directly handle cultures r animals cntaminated r infected with vaccinia virus, recmbinant r defective vaccinia viruses, r ther rthpx viruses that infect humans shuld be ffered the smallpx vaccine. PI s and all ther persns apprved t wrk with the vaccinia virus must be ffered the vaccinia virus vaccine at n charge. Anyne that declines the vaccinia virus vaccine must sign a declinatin frm which will be kept n file with ROHP. Francisella tularensis Bacteria Tularemia, r rabbit fever, is a bacterial disease assciated with bth animals and humans. Accrding t the CDC, Tularemia is a ptentially serius illness that ccurs naturally in the United States. It is caused by the bacterium Francisella tularensis fund in animals (especially rdents, rabbits, and hares). PI s and all ther persns apprved t wrk with F. tularensis must be ffered the tularemia vaccine at n charge. Anyne that declines the tularemia vaccine must sign a declinatin frm which will be kept n file with ROHP. The use f tularemia-causing bacteria at BU/BMC is strictly regulated by the IBC and must be cnducted in designated BSL-3 facilities. Herpes B virus Cercpithecine herpesvirus 1 (Herpesvirus simiae r B-virus) is an endemic infectin in Old Wrld primates f the genus Macaca, which is the species mst cmmnly used in bimedical research. It is assumed that all existing animals in the United States are carriers. Althugh herpes B virus is relatively benign in the macaque mnkey, it can cause rapidly ascending encephalmyelitis in humans with a fatality rate f apprximately 80%. The mst cmmn rute f transmissin f the virus is thrugh bites, scratches, splashes, r cuts. All research invlving nn-human primates must be apprved by bth the IACUC and the IBC. Additinal ccupatinal health screening and persnal prtective equipment (PPE) requirements are described in this manual. Human Tissue and Cell Culture Wrking with Human Tissues and Cells All unfixed human tissue and cells are t be assumed t be infectius (the cncept f universal precautins ) and must be handled using BSL-2 practices and prcedures. Persns wh are expsed t these materials in the labratry are cnsidered t have ptential expsure t bldbrne pathgens, such as human immundeficiency virus (HIV) and hepatitis B virus (HBV), and must be included in the Bldbrne Pathgens prgram. These persns must be ffered the hepatitis B vaccinatin and receive annual bldbrne pathgens training. Page 18 f 201

19 Cell Culture Human r animal pathgens may be assciated with cell r rgan cultures. Cell cultures knwn (r suspected) t cntain an etilgic agent r an ncgenic virus are classified at the same bisafety level as that recmmended fr the agent. The fllwing cell cultures and tissues require BSL-2 r higher cntainment and prcedures: All cultured cells derived frm human surces, including immrtalized and well established cell lines. All cultured cells derived frm nn-human primates, primate lymphid, r tumr tissue. All cultured cells expsed t r transfrmed by a primate ncgenic virus. All clinical materials, such as samples f human tissue btained frm surgery, bipsy, r autpsy. All primate and sheep tissue. All uncharacterized cultured cells new t the labratry until prven t be free f infectius agents. All virus-cntaining primate cultured cells. All mycplasma-cntaining cultured cells. The Bisafety Prgram has develped additinal Agent Infrmatin Sheets that are available nline, fr agents listed n the List f Bilgical Agents with the Ptential t Cause LAI in Use at Bstn University (see Appendix I) Where applicable, agent specific vaccines will be ffered at n charge t all persns wh are apprved by the IBC t wrk with r expsed t these materials. Page 19 f 201

20 Chapter 3 List f Regulatins and Guidelines The fllwing is a summary f federal, state, and lcal agency regulatins and guidelines that either regulate r prvide guidelines cvering the use f bilgical agents: Centers fr Disease Cntrls and Preventin and the Natinal Institutes f Health: Bisafety in Micrbilgical and Bimedical Labratries (BMBL), 5 th Editin, This dcument cntains guidelines fr micrbilgical practices, safety equipment, and facilities that cnstitute the fur established bisafety levels. The BMBL is generally cnsidered the standard fr bisafety and is the basis fr this manual. Natinal Institutes f Health: Guidelines fr Research Invlving Recmbinant DNA Mlecules (NIH Guidelines). This dcument prvides guidelines fr cnstructing and handling recmbinant DNA mlecules (rdna) and rganisms cntaining rdna. Althugh these guidelines are nt subject t regulatry enfrcement, institutins that receive any NIH funding fr rdna research are required t cmply with these guidelines as a cnditin f funding. This dcument requires that each institutin establish an Institutinal Bisafety Cmmittee with the authrity t apprve prpsed rdna research using the NIH guidelines as the minimum standard. Occupatinal Safety and Health Administratin: Bldbrne Pathgens. This regulatin cvers ccupatinal expsure t human bld and ther ptentially infectius materials, including human tissue and cells. OSHA specifies a cmbinatin f engineering cntrls, wrk practices, and training t reduce the risk f infectin. Persnnel ptentially expsed t human bld and ther ptentially infectius material must be ffered immunizatin against hepatitis B and receive annual training. Persnnel wh wrk with HIV r hepatitis B in a research labratry must receive additinal training and demnstrate prficiency in wrking with human pathgens. Bstn Public Health Cmmissin: Recmbinant DNA Technlgy: Use Regulatins (passed March 22, 1994) and Bilgical Labratry Regulatin (passed September 19, 2006). These regulatins require that all institutins in the City f Bstn that wrk with recmbinant DNA mlecules r that perates BSL-3 r BSL-4 labratries be licensed by BPHC. These regulatins require strict adherence t the CDC/NIH guidelines, as well as ther regulatins that the BPHC s Bard f Health and Hspitals may apply. Disease Surveillance and Reprting Regulatin (passed March 30, 2004) requires all institutins in the City f Bstn that engage in research with select agents, Risk Grup 4 agents, and ther agents named by BPHC as high-risk agents t be registered and maintain disease surveillance and reprting prgrams in effect t minimize ptential expsures t these high-risk agents. Cmmnwealth f Massachusetts Department f Public Health: The Center fr Envirnmental Health regulates the strage and dispsal f ptentially infectius material, and includes requirements fr labeling and recrdkeeping. Page 20 f 201

21 Select Agent Rule Department f Health and Human Services: 42 CFR Parts 42 and 43 Pssessin, Use, and Transfer f Select Agents and Txin; Final Rule; and the Department f Agriculture s Animal and Plant Health Inspectin Service: 7 CFR Parts 331 and 9 CFR Parts 121, Agricultural Biterrrism Prtectin Act f 2002: Pssessin, Use, and Transfer f Bilgical Agents and Txin; Final Rule. These regulatins require institutins that pssess, use, r transfer certain bilgical agents and txins ( select agents ) t be registered and apprved by DHHS and/r APHIS. Specific requirements are described in Chapter 10. Other Regulatry Requirements U.S. Department f Transprtatin and the Internatinal Air Transprtatin Authrity: These rganizatins have strict requirements gverning the shipment and transprtatin f hazardus materials, including bilgical agents. Chapter 11 prvides infrmatin n shipping regulatins. Centers fr Disease Cntrl and Preventin: The CDC has established specific regulatry requirements fr imprtatin r transprtatin f etilgic agents, which include a permit applicatin that must be submitted and apprved prir t any such imprtatins. The federal regulatin gverning the imprtatin f etilgic agents is USPHS 42 CFR - Part 71 Freign Quarantine. Part 71.54, Etilgic agents, hsts, and vectrs. U.S. Department f Agriculture, Animal and Plant Health Inspectin Service, and Veterinary Services: USDA, APHIS, and VS regulate the imprtatin f animals and animal-derived materials t ensure that extic animal and pultry diseases are nt intrduced int the United States. Generally, a USDA veterinary permit is needed fr materials derived frm animals r expsed t animal-surce materials. Materials that require a permit include animal tissues, bld, cells r cell lines f livestck r pultry rigin, RNA/DNA extracts, hrmnes, enzymes, mnclnal antibdies fr in viv use in nn-human species, certain plyclnal antibdies, antisera, bulk shipments f test kit reagents, and micrrganisms, including bacteria, viruses, prtza, and fungi. Exceptins t this requirement are human and nn-human primate tissues, serum, and bld. U.S. Department f Cmmerce: The DOC has specific regulatry requirements fr exprtatin f bilgical materials. These regulatins are bth agent and cuntry specific and must be fllwed strictly. Institutinal Bisafety Cmmittee: The IBC has prmulgated a number f specific plicies and prcedures that are incrprated int this dcument as requirements r have been included as appendices. Page 21 f 201

22 Chapter 4 Bisafety Principles The risk f expsure t bilgical agents in a research envirnment depends n a number f parameters (e.g., the agent, its virulence, subject s susceptibility, rute f transmissin, etc.). In general, the bisafety prcedures used are designed t prevent such expsures by cntaining the agents. T prperly design the cntainment, it is imprtant t recgnize the ptential rutes f transmissin fr the given agent. Rutes f Transmissin Skin and Mucus Membrane Cntact Lw-energy prcedures, such as decanting f liquids, pipetting, remval f screw caps, vrtex mixing, streaking agar plates, and inculatin f animals, can result in the generatin f infectius drplets, as well as direct cntact with infectius material. Eye cntact is als cnsidered a rute f expsure. Ingestin Muth pipetting presents the highest risk fr ingestin f infectius material. Splashing f material int the muth and indirect ral expsure thrugh tuching the muth with cntaminated hands can als result in the ingestin f infectius material. Strage f fd r drinks in labratries with bilgical agents, as well as strage f utensils and eating and drinking in the lab, can als result in ingestin f infectius material. Percutaneus Inculatin Use f syringes and needles are cnsidered the greatest risk f expsure thrugh inculatin. Accidental inculatin can als ccur as a result f cuts and scratches frm cntaminated items including syringes used fr animal inculatins, as well as animal bites. Inhalatin Many prcedures have the ptential fr generatin f respirable aersls, including snicatin, centrifugatin, blwing ut f pipettes, heating inculating lps, and changing litter frm the cages f infected animals. Cntainment The term cntainment is used t describe safe methds fr managing infectius agents in the labratry envirnment where they are being handled r maintained. The purpse f cntainment is t reduce r eliminate expsure f labratry wrkers, ther peple, and the utside envirnment t ptentially hazardus agents. The fur elements f cntainment are administrative cntrls, wrk practices, persnal prtective equipment, and facility design. Page 22 f 201

23 Primary Cntainment The prtectin f persnnel and the immediate labratry envirnment frm expsure t infectius agents is prvided by gd micrbilgical technique and the use f apprpriate safety equipment, such as bilgical safety cabinets. Secndary Cntainment Prtecting the labratry s external envirnment frm expsure t infectius materials is accmplished by a cmbinatin f facility design and peratinal practices. The risk evaluatin f the wrk t be dne with a specific agent will determine the apprpriate cmbinatin f these elements Safety Equipment Safety equipment includes bilgical safety equipment, enclsed cntainers, safety centrifuge cups, and ther engineered cntrls designed t minimize expsure t bilgical agents. Bilgical safety cabinets are the mst imprtant safety equipment fr prtectin f persnnel and the labratry envirnment, and mst als prvide prduct prtectin. Safety equipment is mst effective at minimizing expsure when wrkers are trained n the prper use f such equipment and the equipment is regularly inspected and maintained. Bilgical Safety Cabinets (BSC) Prper use f a bilgical safety cabinet (BSC) prvides a high level f cntainment that prtects the peratr frm expsure while prviding sme prtectin frm cntaminatin f the material being handled within the wrk envirnment. Because f its imprtance in prviding cntainment and safety prtectins in the labratry, a BSC is cnsidered ne f the mst critical pieces f safety equipment in bilgical labratries. BSCs are designed t cntain aersls generated during wrk with bilgical material thrugh the use f laminar air flw and high efficiency particulate air (HEPA) filtratin. Three types f BSCs (Class I, II, and III) are used in labratries. Open-frnted Class I and Class II BSCs are partial cntainment devices that prvide a primary barrier ffering significant levels f prtectin t labratry persnnel and t the envirnment when used in cmbinatin with gd labratry technique. The Class I BSC is suitable fr wrk invlving lw-t-mderate risk agents where there is a need fr cntainment, but nt fr prduct prtectin. It prvides prtectin t persnnel and the envirnment frm cntaminants within the cabinet but des nt prtect the wrk within the cabinet frm dirty rm air. The Class II BSC prtects the material being manipulated inside the cabinet (e.g., cell cultures, micrbilgical stcks) frm external cntaminatin. It meets requirements t prtect persnnel, the envirnment, and the prduct. The tw basic types f Class II BSCs are Type A and Type B. The majr differences between the tw types may be fund in the percent f air that is exhausted r recirculated and the manner in which exhaust air is remved frm the wrk area. The gas-tight Class III BSC, r glve bx, prvides the highest attainable level f prtectin t persnnel, Page 23 f 201

24 the envirnment, and the prduct. It is the nly unit that prvides a ttal physical barrier between the prduct and persnnel. It is used with high-risk bilgical agents and when abslute cntainment f highly infectius r hazardus material is required. It is imprtant t nte that hrizntal laminar flw benches must nt be utilized fr wrk with bihazardus r chemically hazardus agents. These units prvide prduct prtectin by ensuring the prduct is expsed nly t HEPA-filtered air. They d nt prvide prtectin t persnnel r the ambient envirnment. Principal Investigatrs are respnsible fr ensuring the prper maintenance f lab equipment. BSCs used as primary barriers must be certified annually by a qualified vendr. Cntact the Bisafety Office infrmatin abut vendrs r ther BSC-related infrmatin. Prper peratin and maintenance f a BSC requires knwledge f hw the system perates, as well as training and experience in effective techniques fr wrking within the cabinet vlume withut cmprmising its functins. Additinal details cncerning the design and use f BSCs are prvided in Appendix C. Tw specialized frms f quality cntrl are strngly recmmended fr all BSCs and are required fr cabinets used t cntain Risk Grup 2 r higher agents: At least daily, r each time the cabinet is perated, the peratr r user shuld bserve the magnahelic gauge and nte its relative psitin. Magnahelic gauges measure the pressure drp acrss the utlet HEPA filter and are imprtant indicatrs f filter integrity and lading. The gauge will typically indicate the same measurement ver a lng perid f time. A significant change in the reading ver a shrt perid f time may indicate clgging r a leaking filter. In such cases, the hd shuld nt be used until the prblem is identified and reslved. If the BSC lcated within a labratry des nt have a magnahelic gauge, users must understand the peratin f the airflw mnitr, cntrls, and alarm settings. Annually, the cabinet shuld be certified by an utside cntractr. EHS des nt ffer this service but can prvide names and telephne numbers f lcal certifiers. The certificatin prcess is quick and relatively inexpensive and ensures that the BSC is meeting its perating specificatins and prviding maximum prtectin. In additin, certifiers prvide service and preventive maintenance fr cabinets and can ften frecast expensive requirements like HEPA filter replacements, allwing PIs t budget fr the event. If BSC recertificatin is required, the recertificatin must be cmpleted befre the current certificatin expires. If the certificatin lapses, the BSC may nt be used fr BSL-2 r higher prcedures until it is recertified. The lab will reprt the lapsed recertificatin t EHS immediately. EHS will infrm the PI and lab wrkers nt t use the BSC and affix a label DO NOT USE and assist the lab t get the BSC recertified. Unless a gd reasn exists fr mre frequent certificatin, a ne-year certificate life is apprpriate. The certificate will generally expire n the last day f the mnth in which the certificatin was perfrmed, ne year later (fr example, a certificate issued n June 2, 2007 will expire n June 30, 2008). Page 24 f 201

25 Persnal Prtective Equipment (PPE) Persnal prtective equipment (PPE) includes safety eyewear, face shields, glves, apprpriate respiratry prtectin, and lab cats. This equipment is used t supplement the cntainment prvided by labratry practices and safety equipment. Nte: PPE is designed t prtect labratry wrkers frm serius expsure t bihazardus materials and shuld be used in cnjunctin with apprpriate engineering and administrative cntrls. At a minimum, staff must use lab cats, safety glasses, and glves whenever there is a ptential fr skin cntact, splash, r aersls. Facility Design The design f a facility is imprtant in prviding a barrier t prtect peple wrking inside and utside the labratry, as well as t prtect peple r animals in the cmmunity frm infectius agents that may be accidentally released frm the labratry. Facility design must be cmmensurate with the labratry's functin and the recmmended bisafety level fr the agent being used r stred. The recmmended secndary barrier(s) will depend n the risk f transmissin f specific agents. Fr example, the expsure risks fr mst labratry wrk in BSL-1 and BSL-2 facilities will be direct cntact with the agents r inadvertent cntact expsures thrugh cntaminated wrk envirnments. Secndary barriers in these labratries may include separatin f the labratry wrk area frm public access; availability f decntaminatin equipment (e.g., autclave*); and handwashing facilities. In BSL-3 facilities, additinal safeguards, such as directinal airflw airlck-cntrlled entry and exiting, a shwer used fr persnnel t shwer ut may be required. As the risk fr aersl transmissin increases, higher levels f primary cntainment and multiple secndary barriers may becme necessary t prevent infectius agents frm escaping int the envirnment. Such design features culd include specialized ventilatin systems t ensure directinal airflw; air treatment systems t decntaminate r remve agents frm exhaust air; cntrlled access znes; an airlck at the labratry entrance; r separate buildings r mdules fr physical islatin f the labratry building itself. *Nte: It is IBC plicy that autclaves used t sterilize bihazardus materials be validated mnthly using a sprulatin test and that validatin recrds be kept (see Appendix D). Bihazardus materials can als be dispsed f in a red bag as medical waste withut autclaving. Bisafety Levels Fur bisafety levels (BSLs) represent cmbinatins f labratry practices and techniques, safety equipment, and labratry facilities. Each cmbinatin is specifically apprpriate fr the peratins perfrmed and the dcumented r suspected rutes f transmissin f the infectius agents, as well as fr the labratry functin r activity. The recmmended bisafety level fr an rganism represents the cnditins under which the agent can be rdinarily handled safely. Page 25 f 201

26 NIH s Guidelines fr Research Invlving Recmbinant DNA Mlecules classifies human etilgic agents n the basis f their relative pathgenicity. Agents are categrized int fur risk grups (RG). As a general rule, a bisafety level shuld be used that matches the highest RG classificatin f the rganisms invlved. Fr example, wrk with vaccinia virus, a Risk Grup 2 (RG2) agent, shuld be cnducted at BSL-2 r higher; simultaneus wrk with E. cli (RG1), Epstein-Barr virus (RG2), and Mycbacterium tuberculsis (RG3) shuld be cnducted at BSL-3. Descriptins f bisafety levels, as well as assigned bisafety levels fr specific rganisms, are cntained in the CDC/NIH dcument, Bisafety in Micrbilgical and Bimedical Labratries (BMBL). The BMBL utlines fur bisafety levels, summarized belw: Page 26 f 201

27 Bisafety Level Agents Practices Safety Equip. Facilities 1 Nt knwn t cause disease in healthy adults; RG1 Standard micrbilgical practices Use basic persnal prtective equipment including labratry cats, dispsable glves and as necessary eye prtectin Open bench tp, sink required 2 Assciated with human disease, which is rarely serius and fr which preventive r therapeutic interventins are ften available; RG2 BSL-1 practice plus: Limited access Bihazard warning signs Sharps precautins Bisafety manual Primary barriers: Class I r II BSCs r ther cntainment used fr manipulatins f agents that cause splashes r aersls f infectius materials. PPE: lab cats; glves; eye/face prtectin as needed. BSL-1 plus: Autclave available 3 Assciated with human disease fr which preventive r therapeutic interventins may be available; RG3 BSL-2 practice plus: Cntrlled access Decntaminatin f all waste Decntaminatin f lab clthing befre laundering Baseline serum Primary barriers: Class I r II BSCs r ther physical cntainment devices used fr all manipulatins f agents. PPE: prtective lab clthing; glves; respiratry prtectin as needed. BSL-2 plus: - Physical separatin frm access crridrs - Self-clsing, duble-dr access - Exhausted air nt recirculated - Negative airflw int labratry 4 Agents are likely t cause serius r lethal human diseases fr which preventive r therapeutic interventins are nt usually available; RG4 BSL-3 practices plus: Clthing change befre entering Shwer n exit All material decntaminated n exit frm facility Primary barriers: All prcedures cnducted in Class III BSCs r Class I r Class II BSCs in cmbinatin with fullbdy, air-supplied, psitive-pressure persnnel suit. BSL-3 plus: - Separate building r islated zne - Dedicated supply/exhaust, vacuum, and decntaminatin systems - Other requirements utlined in BMBL Nte: Cnsult the BMBL fr a mre cmplete descriptin f the fur bisafety levels, as well as recmmended bisafety levels fr specific rganisms. Page 27 f 201

28 In additin t the fur bisafety levels described abve, there are als fur bisafety levels fr wrk with infectius agents in vertebrate animals, referred t as the Animal Bisafety Level (ABSL). ABSL Agents Practices Safety Equipment (Primary Barriers) 1 Nt knwn t Standard animal care As required fr nrmal care cnsistently cause and management f each species. disease in healthy practices, including human adults; apprpriate medical RG1 surveillance prgrams. Facilities (Secndary Barriers) Standard animal facility N recirculatin f exhaust air Directinal air flw recmmended Handwashing sink recmmended 2 Assciated with human disease; RG2 ABSL-1 practices plus: Limited access Bihazard warning signs Sharps precautins ABSL-1 equipment plus primary barriers: cntainment equipment apprpriate fr animal species. PPE: labratry cats, glves, face, and respiratry prtectin as needed. ABSL-1 facility plus: Autclave available Handwashing sink available in the animal rm. Mechanical cage washer used Bisafety manual Decntaminatin f all infectius wastes and f animal cages prir t washing 3 Indigenus r extic agents with ptential fr serius health effects; RG3 ABSL-2 practices plus: Cntrlled access Decntaminatin f clthing befre laundering Cages decntaminated befre bedding remved Disinfectant ft bath as needed ABSL-2 equipment plus: Cntainment equipment fr husing animals and cage dumping activities Class I r II BSCs available fr manipulative prcedures (inculatin, necrpsy) that may create infectius aersls. PPE: apprpriate respiratry prtectin ABSL-2 facility plus: Physical separatin frm access crridrs Self-clsing, duble-dr access Sealed penetratins Sealed windws Autclave available in facility Page 28 f 201

29 ABSL Agents Practices Safety Equipment (Primary Barriers) 4 Dangerus/extic ABSL-3 practices ABSL-3 equipment plus: agents that pse plus: high risk f lifethreatening disease; RG4 Entrance thrugh change rm where persnal clthing is remved and labratry clthing is put n; shwer upn exiting All wastes are decntaminated befre remval frm the facility Maximum cntainment equipment (i.e., Class III BSC r partial cntainment equipment in cmbinatin with full bdy, airsupplied, psitive-pressure persnnel suit) used fr all prcedures and activities Facilities (Secndary Barriers) ABSL-3 facility plus: Separate building r islated zne Dedicated supply and exhaust, vacuum, and decntaminatin systems Other requirements utlined in the text Page 29 f 201

30 Chapter 5 Labratry Bisafety Practices The fundatins f prtective practices in a labratry lie in an individual s labratry experience, technical knwledge, persnal wrk habits, and attitude tward labratry safety. Unlike administrative cntrls, which are behavirs dictated by regulatin r labratry plicy, the term prtective behavir is used t define an innate part f each individual wrker s persnal apprach t the labratry envirnment. As such, prtective behavirs frm the first and mst imprtant line f defense against injury r expsure in the bimedical wrkplace. Basic Labratry Practices Prudent practices and gd techniques are f primary imprtance in labratry safety. Bth are based n sund technical knwledge, experience, cmmn sense, and an attitude f curtesy and cnsideratin fr thers. Techniques and practices spelled ut in detail as Standard Micrbilgical Practices in the CDC/NIH s Bisafety in Micrbilgical and Bimedical Labratries and the NIH s Guidelines fr Research Invlving Recmbinant DNA Mlecules, as well as in the Natinal Research Cuncil s Bisafety in the Labratry - Prudent Practices fr the Handling and Dispsal f Infectius Materials (Natinal Academy Press, Washingtn, D.C., 1989). Many labratry safety text and reference bks als cntain gd infrmatin. At a minimum, the seven basic rules f bisafety, based n the Natinal Research Cuncil s Prudent Practices dcument, shuld be the basis f any persnal labratry wrk ethic. They are nted in Table 1. Bisafety Practice Table 1: Bisafety practices and blcked rutes expsure Rutes f Expsure Blcked 1. D nt muth pipette. 2. Manipulate infectius fluids carefully t avid spills and the prductin f aersls. 3. Restrict use f needles, syringes, and ther sharps t thse prcedures fr which there are n alternatives; dispse f sharps in leak- and punctureprf cntainers. 4. Use lab cats, glves, safety eyewear, and ther persnal prtective equipment. Inhalatin, ingestin, skin, and mucus membrane cntact Inhalatin, skin, and mucus membrane cntact Percutaneus, inhalatin Skin and mucus membrane cntact Page 30 f 201

31 5. Wash hands after all labratry activities, fllwing the remval f glves, and immediately fllwing cntact with infectius agents. 6. Decntaminate wrk surfaces befre and after use, and immediately after spills. 7. D nt eat, drink, stre fds, r smke in the labratry. Skin and mucus membrane cntact Skin and mucus membrane cntact Ingestin, skin, and mucus membrane cntact Labratry Practice and Technique The mst imprtant element f cntainment is strict adherence t standard micrbilgical practices and techniques. Persns wrking with infectius agents r infected materials must be aware f ptential hazards and be trained and prficient in the practices and techniques required fr handling such material safely. The PI is respnsible fr ensuring that labratry persnnel are prperly trained; the PI may delegate the prvisin f training t the labratry supervisr, but the respnsibility remains with the PI. Each labratry shuld develp an peratinal manual identifying specific hazards that will r may be encuntered and specifying practices and prcedures designed t minimize r eliminate risks. Persnnel shuld be advised f special hazards and shuld be required t read and t fllw the required practices and prcedures. A scientist trained and knwledgeable in apprpriate labratry techniques, safety prcedures, and hazards assciated with the handling f infectius agents must direct labratry activities. When standard labratry practices are nt sufficient t cntrl the hazard assciated with a particular agent r labratry prcedure, additinal measures may be needed. The PI is respnsible fr selecting additinal safety practices, which must be in keeping with the hazard assciated with the agent r prcedure. Labratry persnnel safety practices and techniques must be supplemented by apprpriate facility design and engineering features, safety equipment, and management practices. Each labratry will designate a persn as the Labratry Safety Crdinatr. Nte: Althugh each individual is respnsible fr his r her wn safety, the PI has ultimate respnsibility fr ensuring that persns wrking in the labratry are adequately trained and that they fllw the prescribed safety measures. Labratry Husekeeping and Persnal Hygiene With labratry space at a premium, dedicated bench space is a rarity. Fr thse wh d nt have t share wrk space, persnal safety is greatly enhanced by keeping the area neat, clean, and rderly. Injuries and expsures are mre likely t ccur in prly maintained, disrderly areas. Page 31 f 201

32 If wrk space is shared, the imprtance f maintaining a neat, clean area increases significantly. Cwrkers must rely n ne anther t maximize efficiency and safety. Persnal materials shuld be prperly labeled, waste discarded, and the shared space disinfected r cleaned prir t leaving it fr the next user. The fllwing guidelines shuld be bserved in the labratry: Rutine husekeeping ensures wrk areas are free f significant surces f cntaminatin and hazards. Husekeeping prcedures shuld be based n the highest degree f risk t which persnnel and experimental integrity may be subjected. Labratry persnnel are respnsible fr cleaning labratry benches, equipment, and areas that require specialized technical knwledge. Access t exits, sinks, eyewashes, emergency shwers, and fire extinguishers must nt be blcked. The wrkplace shuld be free f physical hazards. Electrical safety is a pririty, especially as it relates t the use f extensin crds. Equipment shuld be prperly grunded. Overladed electrical circuits and the creatin f electrical hazards in wet areas are t be avided. Surfaces shuld be clean and free f infrequently used chemicals, glassware, and equipment. Unnecessary items n flrs, under benches, r in crners shuld be remved. All cmpressed gas cylinders shuld be prperly secured. Persnal hygiene, including prper handwashing techniques, is als a means by which t enhance persnal prtectin in the labratry. Scrubbing immediately after deglving ensures that cntaminatin f the hand by glve micrpuncture r prir expsure is neutralized befre being spread. The labratry is als an inapprpriate place t perfrm persnal csmetic tasks, such as applying makeup, cleaning r trimming fingernails, r brushing hair. These activities prvide new pprtunities fr expsure and cntribute t retrgrade cntaminatin f the labratry envirnment. Universal Precautins Prudent practices ften verlap with a set f practices knwn as universal precautins. The verarching universal precautin espused by the Bldbrne Pathgens (BBP) Standard (see Appendix M fr a list and mre detailed discussin f universal precautins) shuld be adpted by all labratry persnnel. Universal precautins require that all human bld and tissues be handled as thugh they are infectius. Adpting and applying universal precautins t all labratry reagents clearly creates a heightened awareness f ptential risk and adds anther level f cautin t activities invlving reagents. Administrative Cntrls Administrative cntrls are plicies and prcedures designed t assist with the safe handling f ptentially hazardus bilgical materials. They include training, medical surveillance, vaccinatins, access cntrl, etc. Page 32 f 201

33 Bilgical Hazard Infrmatin Labratry wrkers must be knwledgeable f the hazards assciated with the bilgical agents present in the labratry and have hazard infrmatin available t them. The fllwing are surces f hazard infrmatin fr bilgical agents. Micrbial Agents The CDC/NIH s Bisafety in Micrbilgical and Bimedical Labratries (BMBL) has descriptins f bisafety levels and recmmended bisafety practices fr specific bilgical agents. The Canadian Labratry Centre fr Disease Cntrl maintains Material Safety Data Sheets fr micrbial agents ( Txins Islated bilgical txins are chemical hazards, althugh many such txins prduce adverse effects at dses significantly belw that f traditinal labratry chemicals. Labratry use f islated txins falls under the BU Lab Safety Manual and Chemical Hygiene Plans, and Material Safety Data Sheets (MSDSs) must be maintained and available. MSDSs fr a specific txin shuld be btained frm the vendr upn receipt f the txin. Txiclgy textbks, such as Casarett & Dull s Txiclgy, are als gd surces f hazard infrmatin fr txins. Written Standard Operating Prcedures A manual f written standard perating prcedures (SOPs) fr the labratry, in cmbinatin with this manual and the CDC/NIH publicatins Bisafety in Micrbilgical and Bimedical Labratries and Guidelines fr Research Invlving Recmbinant DNA Mlecules, prvide general requirements fr wrking with bilgical agents. Hwever, because these cver relatively general tpics, individual labratries are required t develp labratry-specific SOPs that cver the bisafety cncerns and labratry prcedures fr that particular labratry. Fr example, labratry-specific SOPs shuld address safe manipulatin f specific rganisms, specific expsure cntrl methds, and specific decntaminatin and waste-handling requirements. Appendix W prvides a recmmended standard frmat fr SOPs, and labratries are encuraged t use this frmat t effectively cnvey the bisafety infrmatin (including use f pictures and illustratins). The labratryspecific SOPs d nt need t duplicate the mre general SOPs cntained in this manual r the CDC/NIH dcuments, but shuld serve as supplements. Security and Inventry f Bilgical Agents In recent years, a number f highly publicized incidents invlving bilgical materials have increased bth public cncerns and regulatry versight cncerning the security f bilgical agents. Even thugh many f the agents used in research labratries d nt pse a real risk t health and safety f the wrkers r the public, the perceptin f such risks is f great imprtance. Page 33 f 201

34 At BU and BMC, each PI must develp site-specific criteria that safeguard all bilgical materials, regardless f their risk grup, frm unauthrized remval. It is the PI s respnsibility t ensure that his r her labratry implements sufficient security measures and prcedures t prevent unauthrized access t bilgical agents. Select agents (see Chapter 10) and ther higher-risk micrrganisms and txins must be stred in a lcked cntainer, and a detailed inventry must be maintained with per CDC requirements. In many instances, during the applicatin review prcess, the IBC will review the prpsed acceptable safeguards and either apprve r recmmend enhancements t the prpsed plans. Preventin f Aersls and Drplets Handling f liquids r dry pwders generally is likely t generate aersls r drplets. In practice, highenergy prcedures, such as centrifuging, vrtexing and mixing, tend t prduce respirable aersls that stay airbrne fr extended perids and are small enugh t be inhaled, while lw-energy prcedures, including pening cntainers and streaking plates, prduce drplets that settle quickly n surfaces, skin, and mucus membranes. Utilizatin f Bilgical Safety Cabinets In general, the fllwing guidelines are recmmended when using bilgical safety cabinets (BSCs): The BSC shuld be certified when it is installed r after it is mved, and annually thereafter (fr infrmatin n cabinet certificatin, call (617) at BUMC and (617) at CRC. The magnahelic gauge shuld be checked regularly. This gauge will nrmally run at a relatively fixed value. When it deviates significantly, the cabinet shuld nt be used until the cause f the deviatin has been identified and fixed. Persnnel shuld understand hw the BSC wrks. Persnnel shuld be familiar with the safe and effective use f any UV lamps inside the BSC and use apprpriate precautins t avid UV-related injuries. Nte: EHS has discuraged the use f UV lamps because they have a shrt half-life and are nt generally maintained apprpriately. The BSC s prtective airflw pattern shuld nt be disrupted. Rapid arm mvement, nearby wrkers, and pen labratry drs may disrupt the airflw pattern and reduce the cabinet s effectiveness. Wrk and the necessary materials shuld be planned t minimize the need t exit and reenter the wrk vlume. Accumulatin f materials in the BSC wrk vlume shuld be minimized t reduce turbulence and ensure prper laminar air flw. The BSC shuld be left running whenever the cabinet is in use. Prper disinfectants that avid damaging the cabinet s interir shuld be used. Wrk surface shuld be wiped with 70% alchl befre use. Each item needed fr the planned prcedures shuld be wiped ff and placed in the cabinet. After the wrk vlume is set up, the BSC shuld run fr at least 5 minutes t allw fr stabilizatin f air flw befre any prcedures are begun. Page 34 f 201

35 If a piece f equipment, such as a centrifuge r blender, will create air turbulence in the BSC, it shuld be placed in the back ne-third f the cabinet. All ther wrk shuld be stpped while this equipment is perating. Open flames shuld be avided in the wrk vlume because they create air flw turbulence that may cmprmise sterility. In additin, the heat buildup may damage the HEPA filters. If a flame is necessary, a burner with a pilt light shuld be used. Electric devices, such as lp sterilizers, are ften satisfactry alternatives t pen flames. A pan with disinfectant and/r a sharps cntainer shuld be placed inside the BSC fr pipette/sharps dispsal. Vertical pipette discard canisters n the flr utside the cabinet shuld be avided. Cntaminated and clean items shuld be segregated, and persnnel shuld wrk frm clean t dirty. The bihazardus waste cllectin bag shuld be in a rigid cntainer. D nt blck air flw int the frnt and rear exhaust grilles. Mve arms slwly when remving items frm r intrducing items int the cabinet wrk vlume. Prtect the facility vacuum system frm bihazards by using dual aspiratr flasks in series (A and B) and placing an in-line hydrphbic HEPA filter (C) between the vacuum trap and the surce valve (D) in the cabinet: Nte: EHS requires that the flasks are placed in a secndary cntainer, such as a nalgene tub. All spills in the cabinet shuld be cleaned immediately. Wrk shuld nt resume fr 10 minutes. When wrk is cmplete, all materials shuld be remved frm the BSC and all interir surfaces shuld be wiped with 70% alchl, r ther apprpriate disinfectants. Glves must be remved befre exiting the Bisafety Cabinet, after tuching r handling cntaminated materials. Labratry cat must be remved and hands thrughly washed befre leaving labratry. Utilizatin f Pipettes Pipettes are used fr vlumetric measurements and the transfer f fluids that may cntain infectius, txic, crrsive, r radiactive agents. Labratry-assciated infectins have ccurred frm ral aspiratin f infectius materials, muth transfer via a cntaminated finger, tuching face (eyes, nse, etc) and inhalatin f aersls. Expsure t aersls may ccur when liquid frm a pipette is drpped nt the wrk surface; when cultures are mixed by pipetting; r when the last drp f an inculum is blwn ut. Page 35 f 201

36 The fllwing utlines safe pipetting techniques t minimize the ptential fr expsure t hazardus materials: Never muth pipette. Always use a pipetting aid. If wrking with bihazardus r txic fluid, cnfine pipetting peratins t a bilgical safety cabinet. Always use cttn-plugged pipettes when pipetting bihazardus r txic materials, even when safety pipetting aids are used. D nt prepare bihazardus materials by bubbling expiratry air thrugh a liquid with a pipette. D nt frcibly expel bihazardus material ut f a pipette. Never mix bihazardus r txic material by suctin and expulsin thrugh a pipette. When pipetting, avid accidental release f infectius drplets. Use t deliver pipettes rather than t cntain pipettes, which require blwut. Be careful nt t disldge the residual liquid. D nt discharge material frm a pipette at a height. Whenever pssible, allw the discharge t run dwn the cntainer wall. Place cntaminated, reusable pipettes hrizntally in a pan cntaining enugh liquid disinfectant t cmpletely cver them. Autclave the pan and pipettes as a unit befre prcessing them fr reuse. Discard cntaminated, brken, r intact Pasteur pipettes and brken glass in a sharps cntainer. Dispse f the cntainer prperly when it is, at mst, three-furths full. Pans r sharps cntainers fr cntaminated pipettes shuld be placed inside the BSC, if pssible. Prper prcedures fr dispsal f plastic pipettes are presented in Chapter 9. Utilizatin f Centrifugatin Hazards assciated with centrifuging include mechanical failure and the creatin f aersls. T minimize the risk f mechanical failure, centrifuges must be maintained and used accrding t the manufacturer s instructins. Users shuld be prperly trained and perating instructins that include safety precautins shuld be prminently psted n r near the unit. Aersls are created by activities such as filling centrifuge tubes, remving plugs r caps frm tubes after centrifugatin, remving supernatant, and re-suspending sedimented pellets. A significant aersl hazard can be created if a tube breaks during centrifugatin. T minimize the generatin f aersls when centrifuging bihazardus material, the fllwing prcedures are recmmended: Use sealed tubes and safety buckets that seal with O-rings. Befre use, inspect tubes, O-rings, and buckets fr cracks, chips, ersins, bits f brken glass, etc. D nt use aluminum fil t cap centrifuge tubes because it may detach r rupture during centrifugatin. Fill and pen centrifuge tubes, rtrs, and accessries in a bilgical safety cabinet. Avid verfilling centrifuge tubes t prevent clsures frm becming wet. After tubes are filled and sealed, wipe them dwn with disinfectant. In the event f breakage during centrifugatin, the unit shuld be decntaminated prir t reuse. Always balance buckets, tubes, and rtrs prperly befre centrifugatin. Page 36 f 201

37 Avid decanting r puring ff supernatant; unless the supernatant must be retained, use a vacuum aspiratr with apprpriate in-line reservirs and filters. Wrk in a bilgical safety cabinet when re-suspending sedimented material. Use a swirling rtary mtin rather than shaking. If shaking is necessary, wait a few minutes t permit the aersl t settle befre pening the tube. Small, lw-speed centrifuges may be placed in a bilgical safety cabinet during use t reduce the aersl escape. High-speed centrifuges pse additinal hazards. Precautins shuld be taken t filter the exhaust air frm vacuum lines, t avid metal fatigue resulting in disintegratin f rtrs, and t use prper cleaning techniques and centrifuge cmpnents. Manufacturers recmmendatins must be meticulusly fllwed t avid metal fatigue, distrtin, and crrsin. Avid the use f cellulid (cellulse nitrate) tubes with bihazardus materials. Cellulid centrifuge tubes are highly flammable and prne t shrinkage with age. They distrt n biling and can be highly explsive in an autclave. If cellulid tubes must be used, an apprpriate chemical disinfectant must be used. Utilizatin f Crystats Use f crystats is very cmmn in many research labratries. These devices may pse ptential hazards assciated with sharp cutting edges and cld envirnments and shuld be handled with extra care. The fllwing guidelines shuld be fllwed when using crystats: Frzen sectins f unfixed human tissue r animal tissue infected with an etilgic agent pse a risk because freezing tissue des nt necessarily inactivate infectius agents. Use f freezing prpellants under pressure is nt recmmended with frzen sectins because they may cause spattering f drplets f ptentially infectius material. Apprpriate glves shuld be wrn during preparatin f frzen sectins. When wrking with human r infected animal tissue, cnsider the cntents f the crystat t be cntaminated and decntaminate it frequently with 70% alchl. Cnsider trimmings and sectins f tissue that accumulate in the crystat t be ptentially infectius and remve them during decntaminatin. Defrst and decntaminate the crystat with a tuberculcidal hspital disinfectant nce a week and immediately after use with tissue knwn t cntain bldbrne pathgens, M. tuberculsis, r ther infectius agents. Handle micrtme knives with extreme care. Stainless steel mesh glves shuld be wrn when changing knife blades. Slutins used fr staining ptentially infected frzen sectins shuld be cnsidered cntaminated. Utilizatin f Inculating Lps Flaming inculating lps can result in spatter and the release f aersls and drplets. Use f an electric micrincineratr is the preferred alternative, t minimizing this issue.. Page 37 f 201

38 Use f Absrbent Materials Wrk surfaces shuld be cvered with absrbent paper r diaper sheets t cllect splashes and drips and t minimize the spread f cntaminatin. The absrbent paper shuld be changed at the end f the labratry prcedure as part f the final cleanup, r at least daily during use. Utilizatin f Miscellaneus Aersl-Prducing Devices and Activities Use f any f the devices listed belw results in cnsiderable aersl prductin. Blending, cell-disrupting, and grinding equipment shuld be used in a BSC when wrking with bihazardus materials. Blenders Safety blenders, althugh expensive, are designed t prevent leakage frm the bttm f the blender jar. They prvide a cling jacket t avid bilgical inactivatin and can withstand sterilizatin by autclaving. If blender rtrs are nt leak-prf, they shuld be tested with sterile saline r dye slutin prir t use with bihazardus material. The use f glass blender jars is nt recmmended because f the ptential fr breakage. If they must be used, glass jars shuld be cvered with a plyprpylene jar t prevent spraying f glass and cntents in the event the blender jar breaks. The blender must be perated within a secndary cntainment basin. A twel mistened with disinfectant shuld be placed ver the tp f the blender during use. When pening blenders, be cgnizant f ptential cntaminatin hazards in the frm f drplets that might becme airbrne r fall n the surfaces; liquid residue n the cap; and pssible expansin f the vlume due t aeratin. Befre pening the blender jar, allw the unit t rest fr at least ne minute t allw the aersl t settle. Placing the blender in a BSC will prvide prtectin against airbrne hazards and placement f a tray lined with absrbent pads wuld assist with cntaminatin cntrl. The device shuld be decntaminated prmptly after use. Lyphilizers Depending n lyphilizer design, aersl prductin may ccur when material is laded int r remved frm the lyphilizer unit. If pssible, sample material shuld be laded in a BSC. The vacuum pump exhaust shuld be filtered t remve any hazardus agents r, alternatively, the pump can be vented int a BSC. After lyphilizatin is cmplete, all surfaces f the unit that have been expsed t the agent shuld be disinfected. If the lyphilizer is equipped with a remvable chamber, it shuld be clsed ff and mved t a BSC fr unlading and decntaminatin. Handling f cultures shuld be minimized and vapr traps shuld be used wherever pssible. Page 38 f 201

39 Snicatrs Snicatin is the use f sund-wave energy fr dispersin, disruptin, r inactivatin f bilgical materials, such as viruses. Snicatrs generate sund waves at very high frequencies (~20,000 + Hz range), which is utside nrmal hearing range. The fllwing are hazards assciated with snicatrs: Ampules Nise: Althugh the 20,000-Hz frequency is utside nrmal hearing range, there are ther surces f nise, such as vibratin frm any lse equipment r ther items n the bench r the liquid itself. If the nise levels are high, nrmal hearing prtectin devices shuld be wrn. Aersls: Aersls present a mre serius ptential hazard and must be taken int cnsideratin. Precautins listed fr blenders and lyphilizers shuld be bserved. Opening ampules cntaining liquid r lyphilized culture material shuld be perfrmed in a BSC t cntrl any aersl prduced. Sealed-glass ampules used t stre bihazardus material in liquid nitrgen have explded, causing eye injuries. The use f plyprpylene tubes (cryvials) eliminates this hazard. These tubes are available dust-free r pre-sterilized and are fitted with plyethylene caps with silicne washers. Heat-sealable plyprpylene tubes are als available. Glves must be wrn when pening ampules r cryvials. T pen a sealed-glass ampule, nick the neck f the ampule with a file, wrap it in disinfectantsaked dispsable twel, hld the ampule upright, and snap it pen at the nick. Recnstitute the cntents f the ampule by adding liquid slwly t avid aerslizatin f the dried material. Mix the cntents withut bubbling and withdraw it int a fresh cntainer. Discard the dispsable twel and the ampule s tp and bttm as medical waste. Lp Sterilizers and Bunsen Burners Sterilizatin f inculating lps r needles in an pen flame generates small-particle aersls that may cntain viable micrrganisms. Alternatively, dispsable plastic lps and needles may be used fr culture wrk where electric incineratrs r gas flames are nt available. Cntinuus flame gas burners shuld nt be used in a BSC. These burners can prduce turbulence that disturbs the cabinet s prtective airflw patterns. Additinally, the heat prduced by the cntinuus flame may damage the HEPA filter. If a gas burner must be used, ne with a pilt light shuld be selected. Electric sterilizers shuld als be cnsidered. Page 39 f 201

40 Persnal Prtective Equipment (PPE) Persnal prtective equipment (PPE) must be prvided withut cst t persnnel. Althugh nt a substitute fr the use f BSCs and gd labratry practices, PPE is cnsidered a primary barrier t infectius agents and prper use will reduce the likelihd f infectin. PPE is the least-desirable expsure cntrl methd because its failure results in direct expsure t the agent. PPE is mst effective when used t supplement primary cntrl methds such as bilgical safety cabinets, safety centrifuge cups, and ther cntainment devices. Apprpriate clthing may als prtect the experiment frm cntaminatin The fllwing are cnsidered PPE: Face Prtectin Gggles r safety glasses with slid-side shields in cmbinatin with masks, r chin-length face shields r ther splatter guards, are required fr anticipated splashes, sprays, r splatters f infectius r ther hazardus materials t the face. Wearing cntact lenses is inapprpriate in the labratry setting. Labratry Clthing Labratry cats, smcks, scrub suits, and gwns are cnsidered labratry clthing. Lng-sleeved garments shuld be used t minimize the cntaminatin f skin r street clthes and t reduce shedding f micrrganisms frm the arms. In circumstances where it is anticipated that splashes may ccur, the garment must be resistant t liquid penetratin t prtect clthing frm cntaminatin. If the garment is nt dispsable, it must be capable f withstanding sterilizatin, in the event it becmes cntaminated. Additinal criteria fr selecting clthing include cmfrt, appearance, clsure types and lcatin, antistatic prperties, and durability. Prtective clthing must be remved and left in the labratry befre leaving fr nn-labratry areas. Dispsable clthing shuld be available fr visitrs, maintenance, and service wrkers in the event it is required. All prtective clthing shuld be discarded in the labratry, disinfected, r laundered by the facility. Persnnel must nt launder labratry clthing at hme. Glves Glves must be selected n the basis f the hazards invlved and the activities t be cnducted. Glves must be wrn when wrking with bihazardus and/r txic materials and physically hazardus agents. Temperature-resistant glves must be wrn when handling ht materials, dry ice, r materials being remved frm crygenic strage devices. Delicate wrk requiring a high degree f precisin dictates the use f thin-walled glves. Page 40 f 201

41 Respiratrs When wrking with hazardus materials, the glve shuld verlap the lwer sleeve and cuff f the labratry garment. A lng-sleeved glve r dispsable arm-shield may be wrn fr further prtectin f the garment. In sme instances, duble glving may be apprpriate. If a spill ccurs, hands will be prtected after the cntaminated uter glves are remved. Glves must be dispsed f when cntaminated, remved when wrk with infectius materials is cmpleted, and never wrn utside the labratry. Dispsable glves must nt be washed r reused. Prtectin frm cntact with txic r crrsive chemicals may als be required. Fr assistance in glve selectin, call yur Department Safety Advisr (DSA). Respiratrs are selected based n the hazard invlved and the prtectin factr required. Certain labratry and clinical situatins require respiratry prtectin t prevent inhalatin f infectius agents. Regulatins, as well as gd safety practice, require that persnnel be medically evaluated, specifically trained, and fit tested prir t wearing respiratry prtective equipment. Cntact EHS if respiratry prtective equipment is required r if there are questins abut the respiratry prtectin prgram. Nte: Use f respiratrs requires cmpletin f the OHSA Respiratry Questinnaire fr medical clearance frm the Research Occupatinal Health Prgram (ROHP), and fit-testing by EHS. Ftwear and Miscellaneus Clthing Guidelines Open-ted shes r sandals are nt allwed in the lab. In additin, wearing shrts r ther clthing that expses the lwer legs is generally cnsidered unsuitable in labratries because it increases the ptential fr skin cntaminatin and absrptin f cntaminants. Further guidance n the use f PPE can be fund in the Chemical Hygiene Plan. Strage and Labeling f Bilgical Agents Bilgical agents must be stred using leak prf and sealed cntainer. Cntainers must be clearly labeled with the identity f the agent and shuld include the universal bihazard symbl (see belw) as physical space n the cntainer permits. At a minimum, secndary (r utside) cntainers must include the universal bihazard symbl (identity f cntents is als desirable). Freezers, refrigeratrs, and ther strage areas must als be labeled with the bihazard symbl; exceptins t this plicy will be cnsidered n an individual basis by the IBC. Waste and cntaminated equipment r ther bjects t be decntaminated must als be labeled with the bihazard symbl. Page 41 f 201

42 Universal Bihazard Symbl The OSHA Bldbrne Pathgen Standard specifically requires that cntainers f human bld r ther ptentially infectius material (OPIM), cntaminated waste, and refrigeratrs, freezers, and ther strage cntainers used t stre r transprt bld r OPIM be labeled with the universal bihazard symbl (flurescent range r range-red). Bihazard Labels and Signs Each labratry must have a sign at the entrance that prvides safety infrmatin t visitrs and service persnnel. Rm signs must cntain designatins fr all labratry hazards in use within the labratry (carcingens, acutely txic agents, reprductive hazards, bihazards, radiactive materials, lasers, and magnetic fields). Envirnmental Health and Safety will prepare the signs fr each dr in accrdance with the requirements f NFPA 704 and BSL-3. Bihazard signs will be psted at the fllwing: Entrances t labratries and animal rms that use agents classified as BSL-2 r BSL-3. Cages r animal rms used fr husing animals infected with BSL-2 r BSL-3 agents. Fr a sample f BU dr signage, see Appendix T. Certain ther areas and pieces f equipment within a labratry may als require signs. Refrigeratrs, freezers, cabinets, and ther strage facilities require the bihazard symbl whenever they are used t stre infectius agents f Risk Grup 2 r higher; human bld r bld prducts; unfixed tissues; cell r rgan cultures; bdy fluids; r excreta. Large pieces f equipment fr handling such materials (e.g., centrifuges, bilgical safety cabinets) must be similarly labeled. Page 42 f 201

43 Chapter 6 Labratry Training Training is a critical cmpnent f any integrated bilgical safety prgram. Training is intended t prvide the understanding, technical knwledge, and tls that the trainee can use t imprve his r her daily labratry safety practices. At a minimum, all persnnel wrking with bilgical materials at BU and BMC must have training in the fllwing areas prir t the start f their experiments: Knwledge f this bisafety manual Experimental prcedures t be used Decntaminatin and spill clean-up prcedures Safe handling methds fr any infectius agent and/r recmbinant DNA (rdna) they might be handling Prper methds fr transprting infectius agents and ther bihazardus materials Bldbrne Pathgens Standard (if they wrk with human bld r bld prducts, unfixed tissue, bdy fluids, rgan, r primary tissue and/r samples cntaminated with bldbrne pathgens) Bstn Public Health Cmmissin requirements (see Appendix Q fr a summary) Other specialized training as Bisafety deemed apprpriate by the IBC r the BSO. The PI is respnsible fr ensuring that his r her emplyees receive prper training in the bihazards and cntrls specific t his r her labratry and the safe cnduct f the experimental prcedures t be used. The Bisafety Prgram prvides different types f training assciated with the BU and BMC bilgical, chemical, and radilgical safety prgrams. Each f these has its wn driver and emphasis. The Research Occupatinal Health Prgram is available as a resurce t the IBC, PI s and the BSO t cmplement required training in the areas f bisafety and specific agents. Mandated General Bisafety Training This training is required by law and/r plicy and must be btained thrugh the BSO because f the regulatry aspects that must be included. An example f mandated general bisafety training is initial bldbrne pathgens training and annual retraining (see Appendix M Bldbrne Pathgen Standard). Mandated general bisafety training is required fr all labratry wrkers (faculty, staff, students, and visiting scientists) at BU and BMC. The exact training required fr a particular persn will depend n the hazards t which he r she is expsed. Labratry Safety Training is a training prgram ffered by EHS that is designed fr thse wrking in labratries. New emplyees, faculty, and staff must attend this training prgram immediately fllwing their hiring r as sn as practical and befre beginning labratry wrk. Attendance at new emplyee rientatin des nt fulfill this requirement. Training includes, but is nt limited t, labratry safety practices, bisafety, chemical safety, bldbrne pathgens, and hazardus waste peratins. Page 43 f 201

44 Labratry Safety Training Labratry safety training satisfies the basic cmpetency regulatry requirements fr thse wrking in labs. It des nt satisfy the need fr department-specific training, shipment f infectius agents, select agents, Bisafety Level 3 r 4 wrk, r ther specialized training. Mandated Specific Training Mandated specific training is als required by law and/r plicy. In sme cases, it is administered and tracked by BSO r EHS, which maintains the recrd files. Examples f mandated specific training include nn-human primate users training, agent specific trainings, r ther specific training required by the IBC r BPHC. Individuals wrking in labratries classified as BSL-3 and abve, r wh are ptentially expsed t specific zntic diseases, must als underg training. Training labratry persnnel in the unique hazards, equipment, and prcedures fr a given labratry is the respnsibility f the PI r labratry manager t administer, dcument, and track. This training is mandated and must be prvided by the PI r labratry manager n a peridic basis t all labratry persnnel. Dcumentatin is als required and must include at least the date and duratin f training, name and psitin f the trainer, tpics cvered, and names f the trainees. HIV/HBV Labratry Training Persnnel wh wrk in research labratries that culture, prduce, r therwise perfrm micrbilgical manipulatin f human immundeficiency virus (HIV) r hepatitis B virus (HBV) must receive additinal training beynd the standard bldbrne pathgen training. Prir t wrking with HIV r HBV, labratry wrkers must demnstrate prficiency in standard micrbilgical techniques, and in the practices and techniques specific t the labratry. Additinally, wrkers must have prir experience in handling human pathgens befre wrking with HIV r HBV. Persnnel wh d nt have experience with human pathgens must be trained in the labratry befre wrking with HIV r HBV. Initial training must nt include the use f infectius agents; rather, training and wrk activities shuld be prgressive as prper techniques are demnstrated. Wrkers are permitted t handle infectius agents nly after demnstrating prficiency t the labratry supervisr s satisfactin. Althugh this specialized, labratry-specific training is the labratry supervisr s respnsibility, the training shuld be crdinated with the BSO t ensure prper dcumentatin and recrdkeeping. Packaging and Shipping f Infectius Agents Training Persnnel wh package and ship infectius agents and diagnstic specimens such as micrrganisms, bld samples, and clinical samples fr pathlgical testing are required by federal and internatinal regulatins t receive training every tw years. EHS ffers this training peridically and upn request. Page 44 f 201

45 Select Agents Training Persnnel authrized t use select agents are required t receive training. This training is designed t meet the specific requirements f the 42 CFR 73 requirements and must be cmpleted prir t any individual starting wrk with select agents; in additin there is an annual refresher curse that all authrized individuals must attend in rder t cntinue with their ability t wrk with select agents. EHS and the lab will maintain cpies f the training recrds fr reference. In additin, persnnel authrized by the IBC t wrk with specific agents designated as bilgical agents with the ptential t cause LAI are als required t receive agent specific training (see Appendix I, List f Bilgical Agents with the Ptential t Cause LAI in Use at Bstn University). Cntact the BSO fr mre infrmatin. Bisafety Level 3 Training Specialized BSL-3 training is required fr individuals wh wrk in a BSL-3 cntainment lab r in the BSL-3 bicntainment facility (see Appendix F). Cntact the BSO fr mre infrmatin. Labratry-Specific Training Individual labratries are required t develp specific training fr the particular agents and prcedures that persnnel will perfrm in that labratry. This training shuld be specific t the hazards in the labratry and t each persn s labratry duties. Each persn in the labratry must understand the hazards assciated with the agent and labratry peratins, hw t prevent expsures t bilgical and chemical agents (see Chemical Hygiene Plans), and trained n the labratry standard perating prcedures. Labratry-specific training shuld nt duplicate the general bisafety training, but instead shuld supplement it. Each labratry must maintain training recrds. The recrds shuld include the names f persnnel in the labratry and their mst recent dates f Labratry Safety Training and training prvided specifically by the lab PI r supervisr. Recrds are als updated and maintained in the Bisafety and Chemical Safety Lgbks. The infrmatin will be updated and maintained by the PI r supervisr. Onging training is required as new hazards and prcedures are intrduced int the labratry. The ccurrence f spills, spread f cntaminatin, near misses, etc., als indicate the need fr refresher training. Other Safety Training Persnnel wh utilize hazardus chemicals, radiistpes, r x-ray generating devices must attend additinal labratry safety trainings. Refresher Training All labratry wrkers and certain categries f building ccupants will be subject t peridic mandatry refresher training. The scpe and details f these refresher trainings will be determined by the IBC and will range frm annually (fr high-risk areas, such as BSL-3 and BSL-4 labratries, r thse required by regulatry mandates, such as Bldbrne Pathgen Standard r Select Agents Rule) t every three years Page 45 f 201

46 (fr lw-risk peratins, such as BSL-1). All labratry persnnel will als cmplete the annual Labratry Safety Training. Nnuser Training Individuals emplyed in high cntainment facilities (BSL-3 and BSL-4) but wh are nt wrking with bilgical materials (e.g., administrative staff, facilities, security, etc.) are als required t attend mandatry training prgrams. The scpe and cntent f these training prgrams will be develped based n the needs f each facility. Page 46 f 201

47 Chapter 7 Decntaminatin and Sterilizatin Decntaminatin is a prcess r treatment that renders a device, instrument, r wrk surface safe t handle. A decntaminatin prcedure can range frm sterilizatin by autclave r ethylene xide t simple cleaning with sap and water. Sterilizatin, disinfectin, and antisepsis are all frms f decntaminatin. Sterilizatin is the use f a physical r chemical prcedure t destry all micrbial life, including highly resistant bacterial endspres. Disinfectin eliminates virtually all pathgenic, nn-spre-frming micrrganisms but nt necessarily all micrbial frms n inanimate bjects (wrk surfaces, equipment, etc.). Effectiveness is influenced by the kinds and numbers f rganisms, the amunt f rganic matter, and the bject t be disinfected and chemical expsure time, temperature, and cncentratin. Antisepsis is the applicatin f a liquid antimicrbial chemical t skin r living tissue t inhibit r destry micrrganisms. It includes using germicidal slutins fr swabbing an injectin site n a persn r animal and fr handwashing. Althugh sme chemicals may be utilized as either a disinfectant r an antiseptic, adequacy fr ne applicatin des nt guarantee adequacy fr anther. Manufacturers recmmendatins fr apprpriate use f germicides shuld always be fllwed. General Prcedures Decntaminatin f cultures and bjects cntaminated by bilgical agents is rutinely perfrmed in micrbilgical labratries. Decntaminatin is a vital cmpnent f micrbilgical safety practice and serves t prtect labratry persnnel (as well as thers) frm infectin and the release f infectius rganisms t the utside envirnment (primarily thrugh persn-t-persn transmissin). Decntaminatin f media, wrk surfaces, and equipment is als necessary t prevent cntaminatin f cultured rganisms. Infectius wastes such as liquid and slid will be handled, treated and dispsed accrding t hazardus waste plicies and prcedures. Liquid wastes such as bacterial r viral culture media frm BSL2 labs will be treated with apprpriate disinfectant prir t sink dispsal. Slid wastes frm the BSL2 labratries will be segregated and placed in bihazard cntainers lined with bihazardus waste bags and dispsed as bilgical wastes. This waste is sealed by the labratry and shipped ffsite fr sterilizatin (see Waste Chart psted in the labratry fr mre infrmatin). All wastes frm the BSL3 labratries will be treated befre dispsal frm the labratry (see Chapter 9) Autclaving is the preferred methd fr treating bilgical wastes. A disinfectant shuld be chsen that is apprpriate fr the rganism in use. All liquid bilgical cultures shuld be deactivated with apprpriate disinfectant. All slid bilgical waste shuld be dispsed f in the bihazard waste cntainers available in Waste created in BSL-3 labratries is required t be autclaved prir t remval frm the labratry (see Chapter 9). Page 47 f 201

48 Methds f Decntaminatin The three main categries f physical and chemical decntaminatin are heat, liquid disinfectin, and vaprs and gases. Heat: Wet heat is the mst dependable methd f sterilizatin. Autclaving (saturated steam under pressure f apprximately 15 psi t achieve a chamber temperature f at least 250 F fr a prescribed time) is the best methd f rapidly achieving destructin f all frms f micrbial life. In additin t prper temperature and time, preventin f entrapped air is critical t achieving sterility because f air s pr heat transfer prperties. Material t be sterilized must cme int cntact with steam and heat. Indicatrs f prper autclave peratin (e.g., autclave tape r autclave-sensitive labels) must be used with each lad t visually indicate successful prcessing. Use f autclave tape alne is nt an adequate mnitr f the sterilizatin s success. The Massachusetts Department f Public Health Medical Waste Management Act has specific quality-cntrl requirements fr autclaves used fr sterilizatin f medical waste. Appendix D describes the prcedures fr such tests. Autclaving Liquid disinfectin: A liquid disinfectant (e.g., 1:10 slutin f husehld bleach yielding a final hypchlrite cncentratin f 0.5%) is used t wipe r sak ptentially cntaminated materials fr a perid f time t kill all pathgenic agents present. Each disinfectant requires varying amunts f cntact time. Gas and vapr: Ptentially cntaminated articles are expsed t a sterilizing gas (e.g., ethylene xide, r ETO) r vaprs frm a chemical (e.g., frmaldehyde). Because f the hazardus nature f the gases and vaprs used, this requires specially designed equipment and facilities. Autclaving uses saturated steam under pressure (apprximately 15 psi) t achieve a temperature in the autclave f at least C (250 0 F). Autclaving can be used t destry vegetative bacteria, bacterial spres, and viruses. When decntaminating bihazardus waste, it is recmmended that the temperature in the waste reach a minimum f C fr a minimum f 20 minutes. The ttal prcessing time required t meet these cnditins depends n several lading factrs (see belw); hwever, it is recmmended that a minimum autclave cycle f ne hur be used when decntaminating waste. When using an autclave, the fllwing guidelines shuld be taken int cnsideratin: Bihazardus materials shuld nt be placed in autclaves vernight in anticipatin f autclaving the next day. Autclaves shuld nt be perated by untrained persnnel. Special precautins shuld be taken t prevent accidental remval f material frm an autclave befre it has been sterilized r the simultaneus pening f bth drs n a duble dr autclave. Dry hypchlrite, r any ther strng xidizing material, must nt be autclaved with rganic materials such as paper, clth, r il: Page 48 f 201

49 WARNING! OXIDIZER + ORGANIC MATERIAL + HEAT = POSSIBLE EXPLOSION Three factrs in cmbinatin determine the effectiveness f autclaving: Temperature: an autclave uses steam under a pressure f apprximately 15 psi t achieve a chamber temperature f at least C. Althugh the autclave chamber may reach C, this des nt necessarily mean that the interir f the lad will reach this temperature. Time: a minimum autclave cycle time f 20 minutes at a chamber temperature f C (time des nt begin as sn as the autclave cycle is initiated) is cmmnly recmmended fr sterilizatin f clean items. Hwever, the ttal prcessing time required t achieve decntaminatin depends n several lading factrs, including the lad cntainer (heat transfer prperties); the amunt f water added t the lad; and the weight f the lad. Fr increased lads, an increased cycle time will be required t ensure effective decntaminatin. Cntact: steam saturatin is essential fr maximum heat transfer. Steam must cntact all areas f the lad. Autclave bags and ther cntainers shuld be left partially pen (r therwise permit entry f steam) t ensure adequate cntact. Studies have shwn that adding water t the interir f the bag imprves the time-temperature prfile f the autclave cycle, thereby increasing the autclave s sterilizatin efficiency. Dry Heat Requiring higher temperature and lnger cntact time, dry heat is less effective than mist heat (autclaving). Nevertheless, dry heat is preferable t mist heat fr decntaminatin f anhydrus materials and clsed cntainers because the misture cmpnent f the steam used in an autclave will nt effectively penetrate anhydrus materials and clsed cntainers. The highest dry heat equivalent temperature that these materials will reach in an autclave is C. The highest temperature that material will reach in a dry heat ven will be the actual temperature inside the ven. A temperature f C fr three t fur hurs is recmmended fr decntaminatin f waste using a dry heat ven. Chemical Disinfectin Disinfectin is the decntaminatin f wrk surfaces, equipment, bilgical safety cabinets, and ther inanimate bjects using antimicrbial agents. Several chemical agents are used as disinfectants. Labratry wrkers shuld remember that there are hazards assciated with all f these chemical disinfectants. Inhalatin and skin cntact shuld be minimized, and eye cntact avided. Apprpriate glves and safety eyewear shuld always be wrn when handling these chemicals. Pertinent infrmatin fr sme f the cmmn chemical disinfectants is summarized in table frmat at the end f this chapter. Page 49 f 201

50 Summary f Chemical Disinfectants Effective Against a Disinfectant Alchl (ethyl, isprpyl) Use Parameters cnc.: 70-85% cntact time: min. Vegetative cells Lipphilic viruses Tuberc le bacilli Hydrphilic viruses ± Bacter ial spres Imprtant Characteristics Eye irritant, txic, flammable, inactivated by rganic matter. Ptential Applicatin Surfaces: wrk and equipment Chlrine Cmpunds cnc.: % (cmmercial bleach 0.5%) cntact time: min ± May leave residue; crrsive; skin, eye and respiratry irritant; inactivated by rganic matter; make up at least weekly. Spills, equipment surfaces, instruments, glassware, water baths Quaternary Ammnium Cmpunds cnc.: 0.1-2% cntact time: min. + + Txic, inactivated by rganic matter. Surfaces (wrk and equipment), BSCs, flr maintenance, glassware, instruments Phenlic Cmpunds cnc.: 0.2-3% cntact time: min ± Leaves residue; crrsive; skin, eye and respiratry irritant; txic; inactivated by rganic matter. Surfaces (wrk and equipment), BSCs, flrs, spills, glassware, instruments, water baths Idphr Cmpunds cnc.: 0.47% cntact time: min ± Leaves residue; crrsive; skin and eye irritant; txic; inactivated by rganic matter. Surfaces (wrk and equipment), BSCs, glassware, water baths Frmaldehyde b cnc.: 4-8% ± Leaves residue; skin, eye and Less effective than ther Page 50 f 201

51 Effective Against a Disinfectant Use Parameters Tuberc le bacilli Vegetative cells Lipphilic viruses Hydrphilic viruses Bacter ial spres Imprtant Characteristics (Frmalin) cntact time: min. respiratry irritant; txic (carcingen). Ptential Applicatin disinfectants but can be used fr equipment surfaces, glassware, instruments Glutaraldehyde cnc.: 2% cntact time: min Leaves residue; skin, eye and respiratry irritant; txic. Equipment surfaces, glassware, instruments a: + = very psitive respnse, ± = less psitive respnse. A blank dentes a negative respnse r nt applicable. b: due t its irritating characteristics and status as a carcingen, frmaldehyde shuld nt be used withut gd lcal exhaust ventilatin. Frm Labratry Safety: Principles and Practices, secnd editin, Diane O. Fleming, Jhn H. Richardsn, Jerry J. Tulis, and Dnald Vesley, eds., American Sciety fr Micrbilgy, Washingtn, D. C. Page 51 f 201

52 Revised: September 2011 Chapter 8 Bihazardus Spill Respnse Even with the mst careful planning and implementatin f a research prject, the pssibility f an incident r spill invlving bilgical materials exists. The fllwing prcedures are intended t prvide a planned respnse t such rare events. In any spill scenari, the pririty f actins is determined by the PEP rule - Peple, Envirnment and Prperty. The highest pririty is t prvide aid t injured persnnel and prevent spill area access t thers. Nte: the fllwing are the general requirements and guidelines fr Bihazardus Spill Respnse; each BSL-3 and BSL-4 facility will develp site specific prcedures. Preplanning fr Bihazardus Spill Cleanup All spills f bihazardus materials d nt represent the same risk t persnnel and the envirnment, making each spill smewhat unique. The vlume f a spill is nt necessarily a valid measure f the risks invlved. Fr example, drpping a glass vial cntaining 1.0 ml f lyphilized anthrax spres pses much greater risk t labratry staff than drpping a 10 liter glass bttle f Escherichia cli K-12 culture. Factrs ther than vlume that must be cnsidered in spill risk assessment include: Lcatin (e.g., bihazard cabinet, cuntertp, flr, equipment) Nature (e.g., tip-ver, aerslizing (spray/splash), drp frm a height) Txicity/infectivity f spilled material Vlatility and viscsity f spilled material Other prperties f material (e.g., ph, nrmality, temperature) Nature f affected surfaces (e.g., absrbent, pitted, smth) Cmplicating materials (e.g., brken glass, clthing, mixing with ther materials) Susceptibility f spilled material t neutralizatin/disinfectin Nevertheless, preplanning f spill respnse will lwer the risk f cleaning up a spill and will increase the likelihd that the spill is handled apprpriately. Principal Investigatrs r Labratry Directrs shuld prepare their labratry fr typical spill scenaris expected in the labratry. Labratry wrkers shuld be infrmed f the hazards f the bilgical agents used in the labratry, the risk assciated with these agents during spill scenaris, hw t safely clean up the agents, and hw t prperly dispse f cleanup materials. Page 52 f 201

53 Spill Cleanup Materials Each labratry area shuld have spill cleanup materials available t respnd t the largest spill anticipated fr that area. At a minimum, the fllwing spill cleanup materials shuld be available in the labratry: Glves (thick, chemical-resistant glves r duble pair f thin, nitrile glves are recmmended) Safety gggles and masks r a face shield (strngly recmmended t avid splashes t the nse and muth) Lab cat r smck t prtect clthing and bdy Absrbent pads Disinfectant apprpriate fr the agents used in the labratry Frceps r ther devices t pick up cntaminated material (especially sharps) Sharps dispsal cntainer Autclavable bihazard bags The spill kits distributed by EHS t BU and BMC labratries may nt be adequate fr the respnse t a bilgical spill. Additinal items needed fr the cleanup f bihazardus agents can be maintained in the labratry. Bihazardus Spill Cleanup Risk Assessment Several factrs must be cnsidered when assessing the risk that a spill represents: Vlume and cncentratin f the spilled material The infectius dse f the spilled material and rutes f expsure Lcatin f the spill Degree f aerslizatin f the agent resulting frm the spill Susceptibility f the spilled material t disinfectin Nature f the affected surface(s) and its ability t hide rganisms frm disinfectin Immune status f immediate persnnel As with any spill scenari (bilgical, chemical, r radilgical), the safety f persnnel is the mst imprtant cnsideratin. Cleanup is t begin nly after it is determined that the persnnel wh will clean up the spill have apprpriate knwledge, training, and equipment. Page 53 f 201

54 Bihazardus Spill Cleanup Prcedures The fllwing are general bihazardus spill cleanup prcedures that are apprpriate fr mst spill scenaris; hwever, the apprpriate respnse t any spill is based n an assessment f the risk assciated with that particular situatin. If in dubt, immediately call the Medical Campus Cntrl Center at (617) r the CRC EHS emergency telephne at (617) Bth respnse lines are active 24/7/365. Bihazardus Spills Inside Bilgical Safety Cabinets Wear a labratry cat (dispsable recmmended), safety glasses, and glves (apprpriate fr the bilgical agent and the chemical disinfectant) during cleanup. Allw the BSC t run cntinually during cleanup. Surrund the affected spill area with absrbent material t prevent spread f the spill. Apply disinfectant apprpriate fr the bilgical agent and allw a minimum f 20 minutes cntact time (r as directed by manufacturer s instructins). Alchl r ther flammable liquids are nt recmmended. Wipe up the spill with a dispsable clth r a twel saked with disinfectant. Wipe the BSC s walls and wrk surface, as well as any equipment in the cabinet, with a disinfectantsaked clth. Place cntaminated items in an apprpriate cntainer (bihazard waste bag, sharps cntainer, r autclavable pan with lid fr reusable items) fr autclaving. Allw nn-autclavable items t have a minimum f 20 minutes cntact time with the disinfectant (r as directed by manufacturer s instructins) befre remving them frm the BSC. Remve prtective clthing and place in a bihazard waste bag fr autclaving. Thrughly wash hands and frearms with sap and water. Allw BSC t run fr a minimum f 10 minutes befre resuming wrk in the cabinet r shutting ff the cabinet. Bihazardus Spills in the Labratry, Outside the Bilgical Safety Cabinet If a BSL-1 agent r less than 100 ml f a BSL-2 agent is spilled, the fllwing prcedures shuld be fllwed: Remve any cntaminated clthing and place in a bihazard waste bag fr autclaving, and wash all areas affected by skin cntact with sap and water. Page 54 f 201

55 Wear a lng-sleeved gwn r lab cat (dispsable recmmended), she cvers, safety glasses (face shield als recmmended), and glves (apprpriate fr bilgical agent and disinfectant). Place absrbent pads ver the spill (t absrb liquid), then place a secnd layer f disinfectant-saked absrbent pads ver the spill. Pur additinal disinfectant arund the spill, being careful t minimize aerslizatin, and wrk frm the periphery tward the center, ensuring thrugh cntact between the spill and the disinfectant. Disinfect all items in the spill area. Allw a minimum f 20 minutes cntact time (r as directed by manufacturer s directins) with the disinfectant. Wipe dwn all equipment, tls, etc., with disinfectant. Place cntaminated items in an apprpriate cntainer (bihazard waste bag, sharps cntainer, r autclavable pan with lid fr reusable items) fr autclaving. Remve prtective clthing and place in a bihazard waste bag fr autclaving. Thrughly wash hands, frearms, and face with sap and water. It is recmmended that cleanup persnnel shwer as sn as pssible. If the spill invlves a BSL-3 agent, r greater than 100 ml f a BSL-2 agent, immediately evacuate all persnnel frm the affected area. Wait fr aersl t settle (usually a minimum f 30 minutes) befre entering the spill area. Exceptin: If the labratry is nt under negative pressure, cleanup shuld begin as sn as pssible t minimize the spread f aersls. In additin, the fllwing prcedures shuld be fllwed: Cntact the BUMC Cntrl Center at and ntify EHS immediately fr assistance with the cleanup. Remve any cntaminated clthing and place in a bihazard waste bag fr autclaving and wash all areas affected by skin cntact with sap and water. Wear a lng-sleeved gwn r lab cat (dispsable recmmended), she cvers, safety glasses (face shield als recmmended), and glves (apprpriate fr bilgical agent and disinfectant). Fr cleanup f a BSL-3 agent, a HEPA-filtered respiratr may be required. Place absrbent pads ver the spill (t absrb liquid), then place a secnd layer f disinfectant-saked absrbent pads ver the spill. Pur additinal disinfectant arund the spill, being careful t minimize aerslizatin, and wrk frm the periphery tward the center, ensuring thrugh cntact between the spill and the disinfectant. Disinfect all items in the spill area. Page 55 f 201

56 Allw a minimum f 20 minutes cntact time (r as directed by manufacturer s directins) with the disinfectant. Wipe dwn all equipment, tls, etc., with disinfectant. Place cntaminated items in an apprpriate cntainer (bihazard waste bag, sharps cntainer, r autclavable pan with lid fr reusable items) fr autclaving. Remve prtective clthing and place in a bihazard waste bag fr autclaving. Thrughly wash hands, frearms, and face with sap and water. It is recmmended that cleanup persnnel shwer as sn as pssible. Bihazardus Spills Inside a Centrifuge Clear the area f all persnnel and allw aersl t settle (usually a minimum f 30 minutes) befre reentering the area. Wear a labratry cat (dispsable recmmended), safety glasses, and glves during cleanup. Fr a BSL-3 agent, a HEPA-filtered respiratr may be required. Transfer the rtr and buckets t a BSC fr cleanup. Using an apprpriate disinfectant, thrughly disinfect the inside f the centrifuge, the rtr, and buckets. Discard cleanup materials and prtective clthing as bihazardus waste. Thrughly wash hands, frearms, and ther parts f the bdy with sap and water. Bihazardus Spills Outside the Labratry During Transprt All bilgical agents are t be transprted frm the labratry inside an unbreakable, well-sealed, primary cntainer cntaining absrbent material that is cntained inside a secnd unbreakable, well-sealed leakprf cntainer (see Chapter 11 fr transprtatin guidelines). Bth the primary and secndary cntainers must be labeled with the universal bihazard symbl and the identity f the agent. In the event a transprt cntainer drps and its cntents are spilled, the fllwing prcedures shuld be fllwed: Immediately clear the area f all persnnel and secure the area. Cleanup shuld be initiated as sn as pssible t prevent spread f aersl. Attempt cleanup nly if apprpriate cleanup materials and prtective clthing are available. Ntify the Medical Campus Cntrl Center at (617) r the CRC EHS emergency telephne at (617) Bth respnse lines are active 24/7/365. Nte: Emplyees shuld becme familiar with ther nn-spill emergencies, such as fire and medical emergencies. EHS has develped special emergency flip charts that are lcated in every lab and prvide Page 56 f 201

57 quick references t emplyees. Emplyees shuld review the charts s that in the event f an emergency, they are familiar with their lcatin and cntent. Site-Specific Spill Prcedures BSL-3 and BSL-4 facilities have site-specific emergency respnse and spill respnse prcedures that are part f the facility SOP. Nte: BPHC is ntified f all emergencies in BSL-3 and 4 Facilities. Spill Respnse When respnding t a spill, the fllwing rules shuld be fllwed: Tend the injured: Ensure receipt f immediate medical care and d nt attempt t mve the injured individual(s) unless ambient cnditins becme life-threatening. Individuals splashed, sprayed with, r therwise expsed t human bld r ther bdy fluids r tissues during a spill will need t remve cntaminated clthing and utilize basic first aid, washing any wunds immediately. Await assistance: Unless labratry persnnel are trained and prperly supplied with persnal prtective equipment, DO NOT attempt t clean up the spill. Persnnel shuld immediately call the Medical Campus Cntrl Center at (617) r the CRC EHS emergency telephne number, (617) Bth respnse lines are active 24/7/365. Islate the spill: Evacuate the immediate spill area r the entire rm in the case f an aerslizing (splashing r spraying) spill r a spill f vlatile material. Prevent thers frm entering the spill area with barricades r, if necessary, a sentry. Cntain the spill: Place absrbent material arund, n, r in the flw path f the spilled material nly if it can be dne safely. Prvide infrmatin: Prvide the infrmatin requested by the Cntrl Center r EHS persnnel and await arrival f the emergency prvider. Clean up: Clean up shuld take place ONLY if labratry persnnel are trained, prperly supplied with persnal prtective equipment, and therwise able t clean up and disinfect the spill safely. Page 57 f 201

58 Chapter 9 Bihazardus and Medical Waste Dispsal In the Cmmnwealth f Massachusetts, bihazardus waste is gverned by the Department f Public Health regulatin 105 CMR 480, Strage and Dispsal f Infectius r Physically Dangerus Medical r Bilgical Waste, State Sanitary Cde Chapter VIII. The regulatin defines bihazardus waste as infectius r physically dangerus medical r bilgical waste that because f its characteristics may cause, r significantly cntribute t, an increase in mrtality r an increase in serius irreversible r incapacitating reversible illness; r pse a substantial present ptential hazard t human health r the envirnment when imprperly treated, stred, transprted, dispsed f, r therwise managed. The fllwing types f waste are identified and defined as infectius r physically dangerus medical r bilgical waste, and shall be subject t the requirements f 105 CMR : Bld and bld prducts: Discarded bulk human bld and bld prducts in free draining, liquid state; bdy fluids cntaminated with visible bld; and materials saturated/dripping with bld. Pathlgical waste: Human anatmical parts, rgans, tissues, and bdy fluids remved and discarded during surgery r autpsy, r ther medical prcedures and specimens f bdy fluids and their cntainers. Cultures and stcks f infectius agents and assciated bilgicals: All discarded cultures and stcks f infectius agents and assciated bilgicals, bitechnlgical by-prduct effluents, cultures f specimens frm medical and pathlgical labratries, cultures and stcks f infectius agents frm research labratries, wastes frm the prductin f bilgicals, and discarded live and attenuated vaccines intended fr human use. Cntaminated animal carcasses, bdy parts and bedding: The cntaminated carcasses and bdy parts and bedding f all research animals knwn t be expsed t pathgens. Sharps: Discarded medical articles that may cause puncture r cuts, including but nt limited t all, used and discarded hypdermic needles and syringes, Pasteur pipettes, brken medical glassware, scalpel blades, dispsable razrs, and suture needles. Bitechnlgical by-prduct effluents: Any discarded preparatins made frm genetically altered living rganisms and their prducts. Infectius r physically dangerus medical r bilgical waste shall be referred t as Waste in the subsequent prvisins f 105 CMR Bihazardus Waste Bihazardus waste includes waste materials derived frm cultures and stcks f infectius agents, human pathlgical wastes, cntaminated and nn-cntaminated animal carcasses and bdy parts, all sharps, and human bld and bld prducts. Page 58 f 201

59 Prper handling and dispsal f bihazardus waste is necessary t prevent infectin f persnnel (labratry wrkers, custdians, labratry visitrs, etc.) and release t the envirnment. OSHA and Cmmnwealth f Massachusetts regulatins (105 CMR ) require that bihazardus waste be prperly labeled, stred, and dispsed f. Labeling Bihazardus Waste At a minimum, all bihazardus waste must be labeled with the universal bihazard symbl. Additinal infrmatin, such as the type f waste (such as sharps r liquid waste ) and rigin f the waste, is recmmended. Handling and Dispsal f Bihazardus Waste Sharps Sharps include all syringes, lancets, scalpels, and ther similar medical instruments (whether r nt cntaminated), as well as cntaminated Pasteur pipettes and brken glass, and ther instruments r materials that can cut r puncture persnnel. Sharps must be cllected in rigid cntainers that are leak-prf and resistant t puncture frm the sharps. Sharps cntainers must be designed s that sharps can be safely intrduced int the cntainer but nt easily retrieved. Cntainers shuld be red r range in clr and labeled with the universal bihazard symbl. When the sharps cntainer is apprximately 3/4 full, BUMC/BMC persnnel shuld seal the waste cntainer and it will be picked up by the building facilities custdian r apprpriate service persnnel. CRC persnnel shuld seal the waste cntainer and fill ut a pick-up request n the EHS website ( HS). Waste will be picked up by a waste management cntractr wrking fr EHS. A licensed vendr retrieves the waste frm each building at pre-determined intervals t prcess the waste with an apprved sterilizatin methd. Uncntaminated Labratry Glassware and Brken Glass Cllect uncntaminated labratry glassware and brken glass in rigid cntainers (separate frm ther waste) that will prevent cuts and punctures t persnnel. Cntainers shuld be labeled brken glass. Brken glass is t be dispsed f as rdinary trash. Slid Bihazardus Waste Slid bihazardus waste includes micrbial agents, tissue culture, and cntaminated material (such as petri dishes, pipettes, cntaminated glass, etc.). These materials are cllected in red bihazard bags that are duble-lined and placed in cardbard bxes. BU/BMC persnnel shuld seal the waste cntainer fr pick up by the building facilities custdian r apprpriate service persnnel. Page 59 f 201

60 CRC persnnel shuld seal the waste cntainer and fill ut a pick-up request n the CRC website ( HS) A licensed vendr retrieves the waste frm each building at pre-determined intervals t prcess the waste with an apprved sterilizatin methd. Liquid Bihazardus Waste Liquid bihazardus waste includes all bld and liquid waste frm humans r animals, and all ther liquid bihazardus waste (such as micrbial cultures). Cllect liquid waste in clseable, rigid, plastic, leak-prf cntainers labeled with the universal bihazard symbl. Human and animal bld and bdy fluids can be dispsed f by flushing directly t the sanitary sewer (wear labratry cat, safety glasses and face shield, and glves, and be careful t minimize splashing). All ther liquid waste must be autclaved r treated with a disinfectant prir t dispsal. Liquid waste treated with small quantities f bleach r ther husehld disinfectants can be dispsed f by flushing directly t the sanitary sewer after sufficient cntact time. Liquid waste treated with ther chemical disinfectants must be dispsed f as hazardus chemical waste thrugh EHS. Animal Carcasses, Bdy Parts, and Tissue Infectius animal carcasses are placed in a red bihazard bag secured with a plastic tie and dispsed as bilgical wastes. All nn-preserved carcasses shuld be stred in a freezer r cld strage area prir t dispsal. Secure limbs and sharp prtrusins s they d nt puncture the bag. Animal carcasses frm ABSL3 are treated by autclaving. Validatin f autclaving is dne prir t dispsal as bihazardus wastes. Animal carcasses that are autclaved frm the ABSL4 are dispsed by tissue digestin prcess. Carcasses that are nn-infectius and nt transgenic will be dispsed f thrugh a cmmercial waste dispsal cmpany. A licensed vendr retrieves the waste frm each building at pre-determined intervals t prcess the waste with an apprved sterilizatin methd. Page 60 f 201

61 Chapter 10 CDC/USDA Select Agents The U.S. Department f Health and Human Services (DHHS) and the U.S. Department f Agriculture s (USDA) Animal and Plant Health Inspectin Service (APHIS) regulatins require institutins that pssess, use, r transfer certain bilgical agents and txins (knwn as select agents ) be registered and apprved by DHHS and/r APHIS. DHHS actins are prmulgated under the authrity f the Centers fr Disease Cntrl and Preventin (CDC), and APHIS actins are under USDA enfrcement. The CDC and USDA have identified specific bilgical agents and txins they cnsider t be a severe threat t public health and safety because f their ptential use as biterrrism agents. These materials are referred t as select agents by the CDC and as high-cnsequence livestck pathgens and txins and listed plant pathgens by the USDA. Their transfer, pssessin, use, and dispsal are strictly regulated. This chapter will refer t all these agents as select agents. The current list f select agents is prvided at the end f this chapter. Because the list f select agents may be revised, it is recmmended that the CDC Select Agent Prgram website and the USDA Agriculture Biterrrism Prtectin Act website be checked befre acquiring pathgenic agents and bilgical txins ( The regulatins assciated with select agents are very cmplex and strict, and significant mnetary fines and criminal penalties are assciated with nn-cmpliance. The infrmatin in this chapter is a summary f the select agent regulatins; it is nt a cmplete descriptin f the regulatry requirements. Investigatrs must review and understand the select agent regulatins and their respnsibilities prir t acquiring r wrking with any select agent (fr regulatry infrmatin, see Respnsible Official and Authrizatin Respnsible Official (RO) Select agent regulatins require that a Respnsible Official (RO) be designated fr each institutin that pssesses and uses select agents. The RO has institutinal respnsibility fr the bisafety, security, and regulatry cmpliance f select agents, and as such, must be cntacted prir t btaining any select agents. Authrizatin t Pssess and Use Select Agents Prir t persnnel btaining any select agent, the IBC must review and apprve any prpsed wrk. In additin, PIs wh want t acquire, pssess, r use any bilgical agent r txin listed as a select agent must be registered and apprved with the apprpriate agency (CDC r USDA) prir t btaining the agent(s) r txin(s). The apprpriate agency must apprve bth the institutin, whether BU r BMC, and the individual labratry. Investigatrs wh want t pssess and use select agents must cntact the RO fr assistance Page 61 f 201

62 with the registratin applicatin prcess. Apprval by the CDC r USDA can take several mnths, and PIs shuld plan research prjects accrdingly. The select agent regulatins cntain very strict requirements regarding bisafety, training, emergency respnse, security and accuntability, as well as ther requirements. Investigatrs wanting t acquire select agents shuld thrughly review the CDC r USDA select agent regulatins (whichever is apprpriate) befre initiating the registratin applicatin prcess. Exemptins and Exclusins Diagnstic labs that d nt maintain select agents are largely excluded frm the CDC and USDA select agent regulatins; hwever, ntificatin and pssessin time limits and ther requirements d apply. Additinally, the CDC and USDA can grant exclusins fr temprary public health emergency situatins and ther special circumstances. Cnsequently, any labratry that cnducts diagnstic r verificatin testing fr any select agent must identify itself by cntacting the RO as sn as pssible. Identificatin f any select agent in a specimen r islate must be reprted t the RO as sn as pssible. The RO will assist the diagnstic labratry with reprting t APHIS/CDC within 7 days f identificatin. Labratries must fill ut Reprt f the Identificatin f a Select Agents r Txin (Frm 4) within 7 days f the identificatin. Frm 4 can be fund n the Natinal Select Agents Registry website: Several specific select agent micrbial strains and txin frms have been determined nt t present a severe threat t public health and safety, and are therefre excluded frm the CDC/USDA select agent regulatins. The list f excluded bilgical agents and txins is dynamic and the mst current list is available n the CDC and USDA select agent websites. Txins are excluded based n threshld quantities. Labratries maintaining exempt quantities f select agent txins must keep an accurate inventry f txin amunts t verify that ttal quantities are belw the threshld. Within BU and BMC, each select agent labratry must develp and implement three written plans security, incident respnse, and bisafety that address safety issues assciated with that specific select agent. Security Requirements Security Plan Each select agent labratry must have a written security plan that addresses the fllwing tpics: Physical security Cyber security Inventry f select agents Select agent transfers Training Reprting f unauthrized persns and missing materials Prvisins fr cleaning, maintenance, and repairs Page 62 f 201

63 Any theft r lss f select agents must be immediately reprted t the RO, BUMC Security r BU Plice and EHS, which will ntify the CDC r USDA as apprpriate in additin Bstn Public Health Cmmissin (BPHC), will als be ntified f such events. BU in cllabratin with City Agencies (BPHC, Bstn Fire Department, Bstn Plice Department and EMS) has develped an Emergency Ntificatin and After Actin Guideline High Risk Materials Matrix utlining the list f agencies and events fr which they will be ntified. This plan must be available and up-t-date during a CDC inspectin. The Department f Justice must apprve all persns wh will have access t any select agent. Apprval requires that each individual successfully pass a backgrund security check (cnducted by the FBI in accrdance with the USA Patrit Act) and submit fingerprints t the FBI. Anyne wh has nt been apprved fr access t select agents must be denied access unless escrted by an apprved persn. Everyne wh enters a labratry where select agents are accessible must have security apprval r be accmpanied by an apprved persn. This includes visiting scientists (BU/BMC r ff-campus), maintenance wrkers, custdians, and vendrs. Incident Respnse Incident Respnse Plan Each labratry that pssesses r uses select agents must develp a written emergency plan that is labratry specific and crdinated with department, building, and BU/BMC emergency plans. The plan must address the fllwing: Hazards f the select agents Planning and crdinatin with emergency respnders Building evacuatin, site security, and cntrl Decntaminatin and emergency medical treatment, and ther emergency respnse issues Any expsure r ptential expsure shuld be reprted immediately, and the BU Incident Respnse Plan fr Select Agent Labratries and the Bilgical Spill Respnse Plan fr Select Agent Labratries implemented. Affected persnnel shuld immediately cntact the Research Occupatinal Health Prgram fr medical evaluatin and treatment by calling (617) (ROHP). Labratry persnnel shuld becme familiar with these plans. Bisafety and Recrdkeeping Plan Training f Persnnel All persns apprved fr access t select agents must receive dcumented training cvering the fllwing: Bisafety f select agents and their safe handling, use, and dispsal Security requirements and prcedures Inventry and accunting prcedures Page 63 f 201

64 Emergency respnse prcedures. Training is required befre beginning wrk with select agents and annually thereafter. Transfers f Select Agents Select agents can nly be transferred between entities that are currently apprved by the CDC r USDA t pssess and use select agents. All transfers f select agents (including inter and intrafacility transfers) require prir apprval f the CDC r USDA. Bth the sender and recipient must cmplete a cmmn transfer frm (Frm 2), and the recipient must submit the cmpleted frm t the CDC r APHIS. Frm 2 requires the signature f the RO at bth the sender and recipient facilities. and signature by CDC/APHIS. When the select agent is cnsumed r destryed, the recipient must ntify CDC r APHIS thrugh an update f their registratin. Inventry and Dispsal f Select Agents An accurate recrd f all select agents, frm receipt t destructin r dispsal, must be maintained. The inventry must include specific infrmatin n individual cntainers and vials, as well as a recrd f each use, and ultimate dispsal. The select agent inventry must be verified at least mnthly t accunt fr all quantities and cntainers f select agents. Any discrepancies between the inventry recrd and the actual inventry must be reprted immediately t the RO. Recrds Required fr Select Agents Select agent regulatins require that several recrds be maintained, including the fllwing: Detailed inventry f each select agent and assciated cntainers Access t select agents Access t the area where select agents are used r stred Safety, security, and emergency respnse plans Training recrds Transfer dcuments (Frm 2) Safety and security incident reprts Each individual labratry is respnsible fr maintaining these recrds. EHS will keep recrds f any training that it cnducts. Labratries must maintain recrds f training fr their wrk cnducted by labratry persnnel r any ther applicable training. The Bisafety Office will maintain Frm 2s and any ther CDC r APHIS select agent frms; hwever, labratries must als maintain cpies f these frms. The recrdkeeping requirements are cmplex, and therefre the CDC r APHIS regulatins shuld be reviewed fr a cmplete descriptin f the recrdkeeping requirements. Recrdkeeping fr Select Agent Labratries The fllwing is a list f required recrdkeeping fr all select agent inventry and access recrds, per CDC and APHIS regulatins. Page 64 f 201

65 Lgbk Inventry 1. Name, characteristics, surce data 2. Quantity f first inventry (txins nly) 3. Quantity acquired, surce, date 4. Quantity, vlume, mass destryed r dispsed, date 5. Quantity used 6. Transfers: quantity, date, individual 7. Current quantity held 8. Lst, unaccunted fr, stlen 9. Written explanatin f any discrepancy Access Recrds 1. Access t SA: Name, SA, date remved and returned, quantity used (txins), quantity returned (txins). 2. Access t Area: Name, date, and time entered and left area; uncleared individuals must be accmpanied by apprved individuals and recrded as such. Recrds are reviewed rutinely by the RO and EHS t ensure that they being maintained and updated apprpriately. Recrds must be maintained fr 3 years. Further infrmatin regarding select agents and recrds will be given t labratries wrking with select agents in specialized safety training by labratry persnnel authrized t wrk with such agents. Persnnel, Respnsibilities, and Prcedures fr Select Agent Prcurement and Receipt Prcedure Descriptin This Standard Operating Prcedure (SOP) establishes the BU and BMC prcedures t be fllwed when btaining bilgical agents and txins that have been designated as Select Agents under Federal Regulatin (42 CFR 73.0, 7 CFR 331, 9 CFR 121). This SOP implements the requirements f the BU/BMC Materials Transprtatin Mitigatin Plicy as regards Hazardus Materials including Select Agents. Scpe This prgram cvers all emplyees and staff f Bstn University, including affected PIs, EHS, the Research Occupatinal Health Prgram, and the IBC. Page 65 f 201

66 Respnsibilities Envirnmental Health and Safety (EHS) Manages and versees the acquisitin and receipt f select agents in accrdance with the BU/BMC Materials Transprtatin Mitigatin Plicy. Bisafety Officer (BSO) Prvides guidance t BU/BMC persnnel and assists in cmplying with the requirements f the Public Health Security and Biterrrism Preparedness and Respnse Act f 2002 (Public Law ). Prvides required DHHS, USDA, and APHIS/CDC frms and registratin/reprting dcuments necessary fr btaining apprval t transfer select agents. Cmmunicates directly with CDC Select Agent Prgram t facilitate transfer apprval and t reslve issues. Respnsible Official (RO)/Alternate RO Verifies that each transfer f a select agent cmplies with Public Law Verifies that each transfer is authrized and cmpleted, and cmpletes required dcumentatin. Dcuments successful receipt r reprts theft, lss, r release as required by regulatin. Occupatinal Health Officer (OHO)/Designee The OHO cnducts medical surveillance f emplyees wrking with high-risk agents. The Occupatinal Health Officer is respnsible fr having in place a general plan t determine whether emplyees wrking with varius high-risk agents have had a significant expsure t a high-risk agent, as well as a plan t mnitr significantly expsed emplyees. The OHO is als respnsible fr the existence f a prject-specific plan, created in cnjunctin with the individual labratry supervisr. The OHO, labratry supervisr, the IBC, and the EHS Directr f Research Safety must sign ff n the plan. The Occupatinal Health Officer reprts t the Bstn Public Health Cmmissin any labratry emplyee r individual having access t the labratry wh has been diagnsed r is exhibiting symptms f, r may have been expsed t, any high-risk agent within ne business day. The OHO will cnduct a fllw-up assessment and prvide the BPHC with additinal infrmatin requested regarding islatin and/r quarantine issues. Reprt f unplanned absence due t illness lasting tw r mre days invlving emplyees wrking with high-risk agents must be reprted t BPHC within tw business days. If the illness is believed t be wrk related, the evaluatin must be cmpleted prir t the emplyee returning t Page 66 f 201

67 wrk, and BPHC must be cnsulted three business days prir t the emplyee s expected return t wrk. The Occupatinal Health Officer must reprt any vilatins f labratry prcedures resulting in the release f a high-risk agent. The OHO must reprt t the BPHC any vilatin r breach f any labratry prcedures r any ther incident that the OHO shuld reasnably believe released a high-risk agent beynd the wrk area. See Appendix U fr BPHC Disease Surveillance and Reprting Regulatins. The Occupatinal Health Officer must prvide written ntificatin f any reprts submitted t the BPHC t the EHS Directr f Research Safety. Principal Investigatr (PI)/Authrized Recipient Understands and fully cmplies with all regulatins and BU/BMC prcedures invlving select agents. Executive Directr f Public Safety and Operatins/Designee Maintains security f lcatins determined t be apprpriate fr the receiving, shipping, and strage f designated materials, as well as the screening and examinatin f vehicles, packages, and persnnel. Directr f Emergency Planning and Respnse Prvides recmmendatins related t emergency management planning, training, and respnse crdinatin t EHS, Facilities Management and Planning, the Department f Public Safety, and members f the Emergency Respnse Team. Participates in the develpment and implementatin f emergency respnse plans, exercises, risk reductin initiatives, and risk preventin measures. Serves as the liaisn t the Bstn Mayr s Office f Emergency Preparedness, as well as t the state f Massachusetts and federal emergency management agencies. Institutinal Bisafety Cmmittee (IBC) Reviews and apprves all prjects invlving hazardus bilgical agents and txins, accrding t CDC/NIH and the City f Bstn Public Health Cmmissin guidelines. Fr a current listing f IBC members cntact the IBC Office at (617) Research Occupatinal Health Prgram (ROHP) Prevents, diagnses, and treats persnnel with injuries r illnesses related t expsures r ptential expsures in research and animal care facilities. In cnjunctin with the IBC, prvides medical supprt t wrk invlving hazardus bilgical agents and txins. Page 67 f 201

68 Apprval Prcess Initial Request fr Select Agent PI ntifies the BU/BMC Bisafety Officer that a select agent needs t be rdered at least tw weeks befre anticipated use date. PI btains any applicable USDA permits that may be required fr the interstate transprt f the select agent, and prvides permits t RO. PI cmpletes Sectin A and blcks 30 and 37 f APHIS/CDC Frm 2 and frwards it t the BSO. Bisafety Officer s Review Upn receipt f request t acquire select agent, the BSO: Cnfirms that the PI is an authrized select agent user. Cnfirms that IBC apprvals, training, and inspectin status are valid and current. Cnfirms that apprpriate labratry and strage facilities are available. Fr each transfer, the BSO verifies that the PI has cmpleted Frm 2 accrding t the Guidance Dcument fr the Reprt f Transfer f Select Agents and Txins. If review is favrable, frwards the partially cmpleted frm t the BU/BMC RO. Rejects request if requirements are nt met. Respnsible Official s (RO) Apprval RO r Alternate RO reviews Frm 2 fr accuracy and transmits Frm 2 t the Sender RO. Fr imprted select agents, the RO r Alternate RO wrks with the PI t btain any needed USDA and/r U.S. Public Health Service (PHS) permits and cmpletes APHIS/CDC Frm 2 as per the Guidance Dcument fr Reprt and Transfer f Select Agents and Txins. Federal Apprval Sender receives Frm 2 frm BU/BMC RO/Alternate RO and cmpletes Sectin B and blcks 31-36, then transmits Frm 2 via facsimile t APHIS/CDC. APHIS/CDC crrespnds with the recipient (BU/BMC RO/Alternate RO) and the Sender RO as required t supprt authrizatin f the shipment. APHIS/CDC issues an apprval authrizatin number that is gd fr 30 days. Faxes apprved Frm 2 t Recipient (BU/BMC RO) and Sender RO. Page 68 f 201

69 Shipping Prcess Order Placement The RO receives apprval authrizatin number and cmpleted Frm 2 frm APHIS/CDC and frwards t BSO. RO designates a transprter t be used wh meets the selectin criteria f the BU/BMC Materials Transprtatin Mitigatin Plicy described in Chapter 11. The BSO, upn receipt f the apprved Frm 2 and authrizatin number frm the RO, crdinates transprt f the select agent frm the shipper t BU/BMC using a methd that tracks the mvement f the select agent being shipped and is in accrdance with the BU/BMC Materials Transprtatin Mitigatin Plicy. Crdinates shipment details and tracking with transprter. Sender crdinates shipment details with BU/BMC BSO. Packages, labels, and ships select agent in accrdance with all federal regulatins. Nte: All transprtatin must be in accrdance with the specifics f the BU/BMC Materials Transprtatin Mitigatin Plicy in effect at the time f the transprtatins. Ntificatins The BSO ntifies the fllwing f shipment, date, and time f delivery: RO PI/Authrized User (Recipient) Executive Directr f Public Safety Manager f Emergency Planning and Respnse IBC Chair/IBC Office Occupatinal Health Officer Transprtatin Transprter accepts, stres, lads and delivers package(s) t the BU/BMC apprved lcatin, using apprved access rutes. Transprter reprts any and all vilatins f law, regulatin, and/r plicy. Transprter will cntact the relevant State Plice department having jurisdictin fr any prblem r incident that may ccur during transit with select agents. Receipt Steps Prir t Arrival RO, BSO, and PI verify that apprpriate labratry and/r strage facilities are available. Page 69 f 201

70 RO and Executive Directr f Public Safety r designee review security prcedures fr receipt and transfer prir t the day f scheduled arrival which includes the means fr securing f the lading dck. The Executive Directr f Public Safety r designee will ensure that a BU/BMC public safety fficer prvides escrt fr the RO and subject materials package. Arrival and Receipt Upn the transprter s arrival, the RO and Executive Directr f Public Safety r their designee(s) verify identity f drivers and accuracy f the shipping papers. RO r designee instructs drivers t wait until the package cntents are verified. The public safety fficer will escrt the RO r designee t the apprved select agent labratry. Nte: if the shipment appears t be damaged the delivery truck will be stpped frm mving frm the premises and BU ERT will evaluate the situatin t determine the best curse f actin. Public agencies (e.g. BPHC, BFD, BPD) will als be ntified immediately. The actual curse f actin will depend n whether r nt there is any sign f bvius leakage, the extent f the damage r suspicin that the integrity f the internal packaging cntainer has been cmprmised. The actin taken may include: Over-packing f the package and remval t the labratry t check the cntents t check the integrity f the inner package Leaving the cntainer in place fr further evaluatin by public agencies fr additinal curse f actin Cntainment f any leakage within the transprt vehicle will be achieved by use f apprpriate (i.e. effective fr the agent) absrbent materials and disinfectants. Verificatin f Shipment After dnning apprpriate persnal prtectin equipment and fllwing established labratry prcedures, the PI and RO r their designee(s) take the unpened package t a previusly designated bilgical cabinet (in a labratry apprpriate fr a select agent). The PI and RO r their designee(s) examine the package fr any signs f tampering, damage, r leakage and then pen the package and verify cntents. The public safety fficer will wait utside the labratry while verificatin f the shipment ccurs. If all is in rder (n nn-cnfrmities) and the cntents are verified, the RO r designee finalizes the transprter s shipping papers. Page 70 f 201

71 Ntificatin and Dcumentatin The RO then cmpletes blcks 41 and 42 f Frm 2, faxes, and mails the frm within 24 hurs f receipt t the shipper s RO and APHIS/CDC. The PI enters the select agent int inventry per established prcedure. Discrepancies Ntificatin In the event f a nn-cnfrmity with the shipment, the RO will immediately ntify: 1) The Executive Directr f Public Safety t hld the transprter. 2) Directr f Emergency Planning and Respnse. 3) APHIS and/r CDC, BPHC and lcal law enfrcement and the Department f Justice in the event f theft r lss. Cmplete and transmit APHIS/CDC Frm 3. References APHIS/CDC Guidance Dcument fr Reprt and Transfer f Select Agents and Txins (APHIS/CDC Frm 2): APHIS/CDC Guidance Dcument fr the Reprt f Theft, Lss r Release f Select Agents and Txins (APHIS/CDC Frm 3): CDC Select Agent website: APHIS Agricultural Select Agent website: BU/BMC Materials Transprtatin Mitigatin Plicy Page 71 f 201

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75 HHS Select Agents and Txins Abrin Btulinum neurtxins List f Select Agents* (* Subject t change by DHHS/APHIS at any time) Btulinum neurtxin prducing species f Clstridium Cercpithecine herpesvirus 1 (Herpes B virus) Clstridium perfringens epsiln txin Cccidiides psadasii/cccidiides immitis Cntxins Cxiella burnetii Crimean-Cng haemrrhagic fever virus Diacetxyscirpenl Eastern Equine Encephalitis virus Ebla virus Francisella tularensis Lassa fever virus Marburg virus Mnkeypx virus Recnstructed replicatin cmpetent frms f the 1918 pandemic influenza virus cntaining any prtin f the cding regins f all eight gene segments (Recnstructed1918 Influenza virus) Ricin Rickettsia prwazekii Rickettsia rickettsii Saxitxin Shiga-like ribsme inactivating prteins Shigatxin Suth American Haemrrhagic Fever viruses Flexal Guanarit Junin Machup Sabia Page 75 f 201

76 Staphylcccal entertxins T-2 txin Tetrdtxin Tick-brne encephalitis cmplex (flavi) viruses Central Eurpean Tick-brne encephalitis Far Eastern Tick-brne encephalitis Kyasanur Frest disease Omsk Hemrrhagic Fever Russian Spring and Summer encephalitis Varila majr virus (Smallpx virus) Varila minr virus (Alastrim) Yersinia pestis OVERLAP Select Agents and Txins Bacillus anthracis Brucella abrtus Brucella melitensis Brucella suis Burkhlderia mallei (frmerly Pseudmnas mallei) Burkhlderia pseudmallei (frmerly Pseudmnas pseudmallei) Hendra virus Nipah virus Rift Valley fever virus Venezuelan Equine Encephalitis virus USDA Select Agents and Txins African hrse sickness virus African swine fever virus Akabane virus Avian influenza virus (highly pathgenic) Bluetngue virus (extic) Bvine spngifrm encephalpathy agent Page 76 f 201

77 Camel px virus Classical swine fever virus Ehrlichia ruminantium (Heartwater) Ft-and-muth disease virus Gat px virus Japanese encephalitis virus Lumpy skin disease virus Malignant catarrhal fever virus (Alcelaphine herpesvirus type 1) Menangle virus Mycplasma capriclum subspecies capripneumniae (cntagius caprine pleurpneumnia) Mycplasma mycides subspecies mycides small clny (Mmm SC) (cntagius bvine pleurpneumnia) Peste des petits ruminants virus Rinderpest virus Sheep px virus Swine vesicular disease virus Vesicular stmatitis virus (extic): Indiana subtypes VSV-IN2, VSV-IN3 Virulent Newcastle disease virus 1 USDA Plant Prtectin and Quarantine (PPQ) Select Agents and Txins Pernsclerspra philippinensis (Pernsclerspra sacchari) Phma glycinicla (frmerly Pyrenchaeta glycines) Ralstnia slanacearum race 3, bivar 2 Rathayibacter txicus Sclerphthra rayssiae var zeae Synchytrium endbiticum Xanthmnas ryzae Xylella fastidisa (citrus variegated chlrsis strain) 1 A virulent Newcastle disease virus (avian paramyxvirus sertype 1) has an intracerebral pathgenicity index in day-ld chicks (Gallus gallus) f 0.7 r greater r has an amin acid sequence at the fusin (F) prtein cleavage site that is cnsistent with virulent strains f Newcastle disease virus. A failure t detect a cleavage site that is cnsistent with virulent strains des nt cnfirm the absence f a virulent virus. Page 77 f 201

78 Chapter 11 Transprtatin f Bilgical Materials The packaging and transprtatin f bilgical materials are subject t strict lcal, state, federal, and internatinal regulatins. This is particularly s if the material is transprted thrugh the public dmain, namely, thse radways, airways, and sea lanes accessible t the public. Therefre, at BU/BMC, unless the material is being mved within a specific campus, legal requirements gverning packaging, labeling, and handling must be fllwed. The intent f the packaging and transprtatin regulatins is t prevent accidental expsure f persnnel wh may handle the material during its shipment. Therefre, certain general criteria apply t all pssible transprtatin scenaris. Prir t transprting any bilgical materials, the fllwing cntrls must be in place: Emergency prcedures (e.g., cntact names and infrmatin, spill cleanup, disinfectin prtcls, etc.) must be knwn t the persn carrying the materials. Cntainer must be apprpriate fr the material being transprted. Material must be packed s that it will stay upright during transprtatin. The cntainers must be prperly labeled. Prper prtective clthing must be wrn during the packaging f the material. Hands shuld be washed after handling materials. Open cuts r ther wunds shuld be cvered befre handling the materials. Aersl generatin must be avided when handling and packing the materials. The persn packaging the material must ensure that the exterir surfaces f each package are free f any ptential cntaminatin by the packed material. Transprtatin within a Campus The fllwing requirements must be bserved during the transprtatin f bilgical materials within a campus (e.g., between tw labratries): At a minimum, all labratry materials must be transprted in a secndary cntainer that is gasketsealed, shatterprf, and leak-prf. Materials shuld never be carried in hands r pckets. The secndary cntainer shuld be clseable and easy t decntaminate; an absrbent pad (r similar material) shuld be placed inside the secndary cntainer t absrb any spills. A labratry cat shuld be wrn during transprt. Label infrmatin must include the identity f the bilgical material r agent, the universal bihazard symbl (if the material r agent is in, r abve, Risk Grup 2), and the sending and receiving labratry identificatin (e.g., PI name and rm number). Each individual cntainer must have enugh label infrmatin t identify its cntents. Other infrmatin shuld be n the utside f the package. The cntainer shuld be carried directly t the intended labratry and nt taken t ffices, cafeterias, r ther public r inapprpriate lcatins. Page 78 f 201

79 Upn delivery, the receiving labratry persnnel shuld be infrmed and the material prperly stred. The package shuld be carefully inspected fr signs f leakage r ther cntaminatin and, if necessary, decntaminated befre pening. Transprtatin between Campuses Transprtatin f bilgical samples between campuses (i.e., BUMC and CRC) is subject t the general cnditins described abve. In additin, because the transprtatin takes place thrugh the public dmain, the fllwing ther cnditins apply: All bilgical samples must be packed accrding t Department f Transprtatin/Internatinal (DOT)/Internatinal Air Transprt Assciatin (IATA) regulatins; this includes triple-packaging all samples, even if exempt materials. The specimen shuld be placed inside a primary cntainer with a tight-fitting, leak-resistant tp (e.g., full rund-threaded screw cap with seal r stpper). The primary receptacle r secndary cntainer shuld be labeled with the universal bihazard symbl if it cntains bldbrne pathgen materials, as per required under the OSHA Bldbrne Pathgen standard 1910,1030. The primary receptacle is placed within a secndary (utermst) cntainer that must meet the fllwing specificatins: Shatter- and leak-resistant. Enugh extra space t hld absrbent and cushining materials. arund the primary receptacle. Label infrmatin must include the categry f the infectius bilgical material r agent, (i.e. Categry A, Categry B, r exempt human r animal specimen), and the sending and receiving labratry identificatin (e.g., PI name and rm number). Each individual cntainer must have enugh label infrmatin t identify its cntents. In additin, a sheet cntaining a descriptin f cntents shuld be placed inside the cntainer between the uter and secndary packaging. Any dry ice r ther clant can nw be added between the secndary and uter packaging layers. This clant material shuld be placed in a shipping bx that cntains a cler cmpartment t ensure that the uter bx is nt damaged by misture frm cld packs r ther clants. All required DOT/IATA labeling and marking infrmatin shuld be n the utside f the package. The BU shuttle system must nt be used fr transprtatin f infectius agents r ther bihazardus materials. If the package cntains exempt human r animal specimens, r materials that fall under the Categry B Infectius Substances categry, the package may be mved ver U.S. radways by a member f the labratry. This exclusin, called by the U.S. Department f Transprtatin as exclusive use allws sme materials that are exempt r Categry B Infectius materials t be transprted by a research r clinical labratry persnnel r curier service. This exclusin des nt apply t Categry A infectius substances r ther categries f Dangerus Gds. This individual must have undergne shipping training in the last tw years. This package must fllw all requirements as described abve. Cntact the Office f Research Safety, EHS fr further infrmatin and questins abut this DOT exclusin. The cntainer shuld be shipped directly t the intended labratry and nt taken t ffices, cafeterias, r ther public r inapprpriate lcatins. Page 79 f 201

80 Upn delivery, the receiving labratry persnnel shuld be infrmed and the material prperly stred. The package shuld be carefully inspected fr signs f leakage r ther cntaminatin and, if necessary, decntaminated befre pening. Packaging and Shipping Infectius Agents via Dmestic Flights Occasins d arise when a PI must either ship r receive bilgical materials frm anther institutin. Such activities are gverned by strict federal and internatinal guidelines. The Internatinal Civil Aviatin Organizatin (ICAO) is the United Natins entity that gverns all internatinal civil aviatin matters. The ICAO s Technical Instructins fr the Safe Transprt f Dangerus Gds by Air gvern the shipping f dangerus gds. These technical instructins have been incrprated int U.S. law and are an acceptable methd f transprt in the United States (49 CFR 175). Packaging and shipping bilgical materials invlves certain risks with numerus ptential liabilities. The Internatinal Air Transprt Assciatin s (IATA) Dangerus Gds Regulatins (DGR), latest editin, is the wrldwide gld standard fr shipping. The IATA regulatins apply t all air transprt, bth dmestic and internatinal flights. Fllwing IATA s DGR ensures that a package will als meet U.S Department f Transprtatin requirements fr grund transprt. All respnsibilities fr packaging and shipment f these agents have been assigned t the shipper. Only prperly trained persnnel may ffer infectius materials fr transprt. The fllwing is nly a summary f the requirements fr packaging and shipping infectius agents and des nt cnstitute prper training. Definitins and Applicability Dangerus gds: articles r substances capable f psing significant risk t health, safety, prperty, r the envirnment when transprted by surface r air. Mst infectius r bilgical materials are cnsidered dangerus gds and therefre subject t shipping regulatin. Infectius substances: substances knwn r reasnably expected t cntain pathgens. Pathgens are defined as micr-rganisms (including bacteria, viruses, rickettsiae, parasites, fungi) and ther agents, such as prins, which can cause disease in humans r animals. Fr the purpses f shipping classificatin, infectius substances are brken int tw categries: Categry A: an infectius substance transprted in a frm that, when expsure t it ccurs, is capable f causing permanent disability r life-threatening r fatal disease in therwise healthy humans r animals. Categry B: an infectius substance that des nt meet the criteria fr inclusin in Categry A. Bilgical prducts: thse prducts derived frm living rganisms manufactured and distributed in accrdance with the requirements f natinal gvernmental authrities (e.g., the FDA). They may have special licensing requirements and are used either fr preventin, treatment, r diagnsis f disease in humans r animals, r fr develpmental, experimental, r investigatinal purpses related theret. Page 80 f 201

81 Bilgical prducts manufactured and packaged in accrdance with the requirements f apprpriate natinal authrities; transprted fr the purpses f final packaging r distributin; and fr persnal healthcare use by medical prfessinals are NOT subject t dangerus gds regulatin. Hwever, bilgical prducts nt gverned by natinal authrities and that are knwn r reasnably believed t cntain infectius substances MUST be classified and shipped accrding t dangerus gds regulatins. Exempt Patient Specimens: patient specimens fr which there is a minimal likelihd that pathgens are present are exempt frm mst f the shipping regulatins. Hwever, they must be marked with the wrds exempt human specimen r exempt animal specimen and must be triple-packed as described belw. Cmpletely Exempt Substances: materials that are ttally exempt fr cnsideratin under the shipping regulatins: Substances cntaining micr-rganisms that are nn-pathgenic t humans r animals Substances in a frm s that any present pathgens have been neutralized r inactivated such that they n lnger pse a health risk Envirnmental samples (including fd and water samples) that are nt cnsidered t pse a significant risk f infectin, and Dried bld spts, fecal ccult bld screening tests, bld r bld prducts intended fr transfusin, and tissues r rgans intended fr transplantatin. Classificatin and Identificatin The substance t be shipped must be classified as cmpletely exempt frm regulatin, an exempt patient specimen, r a Categry A r B infectius substance. Once classified, prper shipping names and identificatin numbers can then be assigned t the material. Exempt patient specimens d nt require shipping names and identificatin numbers. Hwever, Categry A and B materials are assigned the fllwing names and numbers: Categry A: assign ne f tw identifiers, depending n whether r the material infects humans: Infectius substance affecting humans: UN 2814 Infectius substance affecting animals: UN 2900 Nte: If a material infects bth humans and animals, use the Infectius substance affecting human cde, UN Categry B: bilgical substance categry B: UN 3373 Packaging All regulated infectius substances, including Categry A, Categry B, and exempt patient specimens, must be triple packed: The innermst primary receptacle(s) is leak-prf. A leak-prf secndary receptacle with absrbent material placed between the primary and secndary receptacles t prevent the release f liquid during transprt and t shield multiple primary receptacles frm cming in cntact with ne anther. Rigid, tertiary uter packaging that is at least 100 mm (4 in) in its smallest external dimensin. Additinally, shipments f Categry A and Categry B materials must be packaged accrding t IATA Page 81 f 201

82 Packing Instructins 602 and 650, respectively. Thse guidelines require the fllwing: Shipments must be prepared in such a way that they arrive at their destinatin in gd cnditin and present n hazard t persns r animals during shipment. Outer packaging must meet structural strength requirements and carry defined specificatin markings. Packages must be at least 100 mm (4 in) in their smallest external dimensin. An itemized list f cntents must be enclsed between the secndary cntainer(s) and the uter packaging. All packages cntaining infectius substances must be marked durably and legibly n the utside f the package with the name and telephne number f a persn respnsible fr the shipment. The shipper must make advance arrangements with the recipient and the peratr t ensure the shipment can be transprted and delivered withut unnecessary delay. Substances shipped at ambient temperatures r higher must be in primary receptacles made nly f glass, metal r plastic, with a psitive means f ensuring a leak-prf seal. Screw caps must be reinfrced with adhesive tape. Substances shipped refrigerated r frzen must carry the refrigerant between the secndary cntainer and uter packaging. Wet ice is nt recmmended fr shipping as it may cause the package t leak during transprt, thus delaying r causing rejectin f the package by the transprter. If dry ice is used, the packaging must permit the release f CO 2 gas. Primary and secndary cntainers must retain their integrity acrss the full range f pressures and temperatures experienced under nrmal and lss-f-refrigerant cnditins. Page 82 f 201

83 Labeling Package labeling is in the frm f standardized pictures that must be affixed t the utside. The clr and design f each label is prescribed in the IATA regulatins. All labels must be at least 2 inches n the smallest side. Fr the purpses f infectius substances, five different labels must be cnsidered: Categry A: Categry B: UN 3373 Dry Ice: Carg Aircraft Only: must be affixed if shipping vlumes greater than 50 ml f a Categry A substances Orientatin Arrws: if shipping liquids, tw such labels must be affixed t the package, n ppsing sides. Page 83 f 201

84 Marking Markings are the wrds and numbers required t be n the utside f a package. The fllwing markings must be present n any package cntaining a Categry A r Categry B material: UN Number and Prper Shipping Name: UN 2814 Infectius substance affecting humans UN 2900 Infectius substance affecting animals Bilgical substance categry B UN 3373 Nte: The UN number is part f the label fr Categry B substances. Cntact Infrmatin Name and telephne number f the respnsible persn 24-hur emergency telephne number in case f transprtatin emergency T and frm infrmatin If shipping a material under dry ice, the fllwing additinal marking is required: UN 1845 Dry Ice (the weight in kilgrams f the dry ice present shuld als be nted) If shipping an exempt patient specimen, the nly marking required is: Exempt Human Specimen r Exempt Animal Specimen Training Requirements Thse invlved in the packaging and shipping f infectius substances must underg training every tw years r when activities change. It is the department s respnsibility t ensure training is cmpleted. The Office f Research Safety, EHS can prvide this training. The shipper is bligated t receive further qualificatin when shipping hazardus materials f a class r divisin where current training is insufficient. Shipping Dcuments Shipping papers describing the material in transit must accmpany all shipments f dangerus gds. Fr grund transprt, a Bill f Lading is required. Fr air transprt, an Airway Bill takes the place f a Bill f Lading. Hwever, air transprt f a Categry A material als requires that a Shipper s Declaratin f Dangerus Gds be filled ut. The full and accurate cmpletin f the Shipper s Declaratin is essential, as these are legal dcuments signed by the shipper, which creates a cntract between the shipper and the carrier. The dcument must be accurate, legible, and neat and withut any spelling errrs. The declaratin frm must be cmpleted in English. Page 84 f 201

85 Three cpies f the declaratin must be cmpleted. One cpy will remain with the shipper (PI). Tw cpies will be sent with the shipment. If the declaratin is nt a three-part NCR frm, phtcpies must be made. S INSTRUCTIONS 1 Shipper s: Name Address Phne number 2 Receiver s: Name Address Phne number 3 Line ut the item that des nt apply. Passenger aircraft can nly be used t ship quantities less than 50 ml. Carg aircraft must be used t ship quantities between 50 ml and 4 L. 4 Line ut the item that des nt apply. 5 Prper Shipping Name (infectius substance, affecting humans r infectius substance, affecting animals) Identify the specimen by name in parenthesis ex. Infectius substance, affecting humans (rabies virus) 6 Class r Divisin * Always UN Cde * UN 2814 r UN 2900 (UN 3373 des nt require shippers dec.) 8 Packaging Grup * There is n packaging grup fr bilgical agents. 9 Identify by stating the number f cntainers by the quantity in each cntainer. (e.g., 5 X 10ml) Identify type f uter cntainer fr the shipment 10 Packaging Instructins * 602 r 650 (als 904 if dry ice included) hur emergency cntact number fr the shipper (PI, Lab Supervisr), The statements, Prir arrangements as required by the IATA Dangerus Gds Regulatins have been made. And Prepared accrding t ICAO/IATA. 12 Name and Signature f the shipper. * As described in the latest editin f the IATA Dangerus Gds Regulatins Nte: When shipping bimaterials n dry ice, remember that dry ice is itself cnsidered a dangerus gd and must als be listed n the shipping dcuments as UN1845, Packing Grup III, Packing Instructin 904. BU/BMC have adpted additinal shipping requirements fr high hazard materials with strict Page 85 f 201

86 requirements fr apprved carriers, including a dedicated vehicle, pint-t-pint delivery, and specified shipping rutes. Fr mre infrmatin, cntact EHS. The transprtatin must als meet the NIH guidelines. BU/BMC Materials Transprtatin Mitigatin Management Plicy 1. Purpse and Applicability The purpse f this plicy is t define the prcedures used t manage the shipping, receiving, and transprtatin f items determined t be high-risk by Envirnmental Health and Safety in accrdance with Bstn University (BU/BMC) plicies and prcedures and all applicable laws and regulatins. This plicy applies t all items determined t be high -risk and t all emplyees and staff, including thse wh are visiting users f BU/BMC facilities and thse wh are cntracted services invlved in the shipping, receiving, handling r ther use f subject materials as described belw. This plicy defines the prtcls fr the selectin f cntracted services t be used in the shipping, receiving, and transprt f subject materials. It als includes standards fr packaging, transprting, delivery rutes and the quality cntrls t be utilized t ensure that all thse invlved in the management f subject materials transprt adhere t these standards. 2. Definitins Subject Materials: A substance r material in a quantity and frm that may pse a high level f risk t health, safety r prperty when received, transprted and/r stred. These materials include, but are nt limited t, txic/infectius substances (including select agents), radiactive materials, chemicals, cmpressed gases, and any ther materials that Envirnmental Health and Safety (EHS) deems a material that shuld be managed thrughut its transprt. Select Agents: Bilgical agents and txins that have the ptential t pse a threat t public health and safety if used fr biterrrism purpses. The list includes ver 80 bacteria, viruses, txins, rickettsia, and fungi. The prgram is regulated by the Department f Health and Human Services (DHHS) and Department f Agriculture (USDA) under the Federal Regulatin fr Select Agents [42 CFR 73.0; 7 CFR 331; 9 CFR 121]. Shipper: The shipper is the persn wh packages the subject material and signs the shipper s declaratin frm. This persn is respnsible fr the material t be classified, identified, packaged, marked and labeled, with all apprpriate dcumentatin included with the package. This individual is required t have shipping training, and ntify the receiver regarding the planned shipment f highrisk material. Transprter: The transprter is the individual, peratr r cntracted service that btains the package frm the shipper, verifies it has been packaged crrectly, and carries the package t the receiver. Receiver: The receiver, fr the purpses f this plicy, is the individual wh receives the package. This individual is required t have shipping training. The receiver ntifies the shipper upn receipt Page 86 f 201

87 f the planned delivery f high-risk material. Shippers Declaratin Frm: The dcumentatin that a high-risk material will be shipped. These dcuments will be maintained in accrdance with all laws, regulatins and BU plicies including standards fr the maintenance f riginal frms t be maintained by the shipper, the transprter and the receiver. Qualified Vendr: A vendr wh meets r exceeds the criteria in Sectin Rles and Respnsibilities 3.1 Envirnmental Health and Safety The Envirnmental Health and Safety (EHS) is respnsible fr the management and versight f the Materials Transprtatin Management Plicy and fr ensuring cmpliance with the prcedures utlined within this plicy by all emplyees and staff, visiting users f BU facilities and cntracted services including assciated transprters. 3.2 Office f Mail Services The Office f Mail Services will prvide supprt t EHS and Department f Public Safety with the screening/examinatin f delivered packages, with the staffing f designated lcatins, and with the management f cntracted services. 3.3 Office f Purchasing Services The Office f Purchasing Services will be respnsible fr facilitating the selectin f cntracted service prviders wh are capable f prviding services in accrdance with this plicy and in cmpliance with all applicable laws and regulatins. The Office f Purchasing Services will select, mnitr, manage and discharge all cntracted services that are invlved in the management and transprt f subject materials. 3.4 Department f Public Safety The Department f Public Safety (DPS) will, thrugh its Investigatins Unit, initiate, cnduct and/r participate in audits and cnduct investigatins as necessary. DPS, thrugh its Systems and Operatins Units, will be respnsible fr maintaining the security f lcatins determined t be apprpriate fr the receiving, shipping and strage f designated materials as well as the screening and examinatin f vehicles, packages and persnnel. DPS will prvide security at the pint f receipt f the high hazard material and escrt the package frm the pint f entry t the final destinatin in the BUMC. Transprt f select agents frm ne lcatin t anther utside f a cntained area may require security escrt t verify that the transprter is BUMC select agent authrized. 3.5 Office f Emergency Planning and Respnse The Office f Emergency Planning and Respnse (OEPR) will prvide t Envirnmental Health and Page 87 f 201

88 Safety, Facilities Management and Planning, the Department f Public Safety and members f the Emergency Respnse Team recmmendatins related t emergency management planning, training and respnse crdinatin. In additin, the OEPR will participate in the develpment and implementatin f emergency respnse plans, exercises, risk reductin initiatives and risk preventin measures; and serve as the liaisn t the Bstn Mayr s Office f Emergency Preparedness, the Massachusetts and Federal Emergency Management Agencies. 3.6 The Shipper The Shipper will be respnsible fr ensuring that the material being shipped is apprpriately packaged including classifying, identifying, marking, labeling and prviding apprpriate dcumentatin with the package. The shipper must be trained in accrdance with all applicable laws, regulatins and BU plicies including thse that address the type and frequency f training and necessity f additinal training shuld laws, regulatins r BU plicies change at any time. 3.7 The Transprter The Transprter will be required t d the fllwing: accept, stre, lad, inspect and deliver packages t an apprved lcatin using apprved access rutes; reprt any and all vilatins f law, regulatin r plicy; retain all recrds; and have prper shipping training. The inspectin f packages includes requirements invlving damage t packages, reprting guidelines and immediate cmmunicatin t the shipper and receiver, public health and regulatry authrities. In additin t these requirements, transprt cmpanies may have their wn specific safety requirements fr subject material transprt. 4. Prcedures EHS and DPS will determine the best lcatin fr the receipt, cntrl, audit, transprt, and shipping f all items under this plicy. Such lcatin(s) will be perated r prvided with versight by representatives f EHS and ther related user departments. These areas will be rutinely audited. Transprt t and frm this lcatin will be by majr rutes f travel that immediately brder BUMC and are limited t Albany Street, Massachusetts Avenue and the highway/cnnectr system in the rear f BiSquare. EHS will train all users f the laws, regulatins, plices and requirements invlved in the shipping and receiving f subject materials and will manage the tightly cntrlled, pre-apprved, scheduling f shipment and delivery times. EHS will train all BU users in the apprved prcedures fr the packaging f materials, the apprved cntracted services t be used in the transprt f such materials and the penalties f failing t fllw all aspects f this plicy. EHS and DPS will ensure that BU staff invlved in the high-risk materials shipping / receiving areas underg a backgrund clearance check, as apprpriate, cnsistent with the select agent regulatry requirements prir t being apprved t wrk in these lcatins. EHS and DPS will determine the packaging requirements t be used in the shipping and receiving f subject materials. These requirements will cmply with all applicable regulatry standards. Page 88 f 201

89 These mandated packaging requirements wuld nly be altered after btaining any required apprval frm all relevant regulatry authrities. Transprt f select agents will be dne in accrdance with all laws and regulatins including the apprval frm the U.S. Department f Health and Human Services/ Center fr Disease Cntrl and Preventin (HHS/CDC) r United States Department f Agriculture/Animal and Plant Health Inspectin Service (USDA/APHIS), prir t shipment, and ntificatin within 24 hurs f receipt. The transprt will als include the utilizatin f apprpriate frms and the reprting f registratin numbers f all parties invlved in shipping, transprting and receiving packages. EHS, DPS and the Office f Purchasing will select cntractrs fr the transprtatin f subject materials based n criteria including, but nt limited t, the fllwing: Past perfrmance n similar cntracts. Ability t prvide services as a qualified vendr fr transprt f all subject materials. Ability t prvide transprt services in accrdance with all applicable regulatry standards. Ability t prvide transprt services in accrdance with all applicable BU standards. BU requires that the DOT-cmpliant triple packaging be placed in a nn-crushable liquid tight slid cntainer fr an added layer f safety. BU requires that packages be secured in the vehicle away frm ptential impact n uter walls. Ability t prvide staffing that has undergne, and cntinues t underg n an annual basis, apprpriate backgrund checks. Ability t prvide curier services that may require that a single individual pick up and deliver packages. Ability t prvide GPS tracking f packages r vehicles as determined apprpriate and apprved by BU. Ability t prvide vehicles that are inspected in accrdance with all applicable inspectin standards at least every six mnths. Ability t prvide custmized services that require adherence t BU determined rutes f travel, audit prcedures and strictly defined schedules fr bth pick-ups and deliveries. Ability t maintain and t prvide an all-inclusive chain f custdy dcument upn delivery f each package. Ability t prvide resurces t participate in BU audits f services. Page 89 f 201

90 Any transprtatin vendr persnnel having relative prximity t the package must reprt all ccurrences f illness t the Bstn Public Health Cmmissin fr a perid f three weeks frm the delivery departure date. Tracking Shipments: EHS will schedule all deliveries and will track the delivery with the cntracted service perfrming the transprtatin by means f cntractr-prvided tracking methds. BU will initiate its wn tracking methds at its discretin and will determine the type f packaging that the shipper, receiver and transprtatin cmpany uses, and that it is in cmpliance with all laws and regulatins. Prir t the transprt f a shipment t the NEIDL f a select agent, the Directr f Emergency Planning and Respnse will ensure that the apprpriate Cmmnwealth and Bstn emergency respnse departments having jurisdictin are ntified. Off Peak Delivery: EHS will schedule all deliveries t arrive at ff peak traffic hurs thrugh the City f Bstn t ensure transprt and reduce the pssibility f accident r delay due t traffic cngestin. Clear Lading Dck: DPS will ensure that the lading dck r ther facility where the transprter is delivering the high hazard material is free and clear f all parked vehicles t enable safe, secure transfer and receipt. Areas used fr deliveries will include secure lading r vehicle inspectin areas in which the delivery vehicle can be islated frm mvement. Delayed Receipt: Failure t receive package within the specified time range f delivery will result in an immediate investigatin invlving the transprt cntractr, the shipper, BU and all applicable regulatry persnnel. Receipt f Packages: Packages delivered t BU will be inspected, verified, dcumented and transprted t the apprpriate lcatin within BU by EHS. Prir t receipt f the package, the Respnsible Official will verify with the driver that the package s integrity is intact. In the event that the package s integrity is cmprmised, the transprt cmpartment will be sealed and the transprter s emergency prtcls will be fllwed. OEPR will ntify all apprpriate lcal respnse agencies and initiate the BU Emergency Respnse Plan, the BU Select Agent Incident Respnse Plan, and Incident Cmmand System. Prblems r Incidents On Rute: The transprter will cntact the relevant law enfrcement agency having jurisdictin fr any prblem r incident that may ccur during transit r transprt f the subject material. The transprter will als ntify BU immediately f any such event. The transprter will ask that the public safety agency having jurisdictin ntify the lcal emergency respnders having jurisdictin where any incident ccurs. Upn ntificatin f an incident enrute t BU, OEPR will ensure that the lcal emergency respnse departments having jurisdictin are ntified. The transprter will have a reputable hazardus materials cleanup cntractr available n a 24- Page 90 f 201

91 hur by seven days a week basis fr respnse fr a bilgical incident mitigatin. The cntractr will crdinate thse mitigatin effrts with the lcal emergency respnder incident cmmander. Ntice f Successful Transprt: Upn the successful receipt f a shipment under this plicy, OEPR will ntify all the apprpriate public safety agencies f the cnclusin f the transprt. 5. Key References and Resurces U.S. Department f Health and Human Services, Bisafety in Micrbilgical and Bimedical Labratries, 5th Editin, December 2009 U.S. Department f Transprtatin, 49 CFR Part 171 Final Rule, 03/18/05 Current Revised Internatinal Air Transprt Authrity, Dangerus Gds Regulatins U.S. Public Health Service (HHS)/ CDC 42 CFR Part 73.0, Pssessin, Use & Transfer f Select Agents and Txins, 03/18/05 Mrbidity and Mrtality Weekly Reprt Vl. 1 N. RR-19, Labratry Security and Emergency Respnse Guidance fr Labratries Wrking with Select Agents 12/06/02 Natinal Institutes f Health Guidelines fr Research Invlving Recmbinant DNA Mlecules 6. Web Sites BU, Envirnmental Health and Safety (EHS): BUMC, Public Safety Department: BU, Purchasing: Centers fr Disease Cntrl and Preventin: CDC Select Agent Prgram, Labratry Registratin: Federal Express, Dangerus Gds Prgram: Internatinal Air Transprt Authrity: MA Department f Public Health-State Lab Institute: United Parcel Service, Hazardus Materials Supprt Center: USDA Select Agent Prgram: United States Pstal Service: United States Public Health Service: Page 91 f 201

92 U.S. Department f Transprtatin: Page 92 f 201

93 Appendix A IBC Applicatin Frms / Bilgical Use Authrizatin (BUA) Bilgical Use Authrizatin (BUA) IBC Applicatin Frm in RIMS can be fund at: Page 93 f 201

94 Appendix B Imprtatin and Exprtatin f Etilgic Agents Multidisciplinary and multi-institutinal research is a cmmn practice that invlves cllabratin amng faculty frm varius institutins and cuntries. At times it is necessary t share bilgical samples r materials with cllabratrs. Federal regulatins strictly cntrl the imprtatin and exprtatin f etilgic agents. The fllwing utlines tw majr requirements that must be fllwed. Nte: All imprtatin and exprtatin f etilgic agents must be prcessed thrugh the Bisafety Prgram. CDC Etilgic Agent Imprt Permit Prgram Etilgic agents are thse micrrganisms and micrbial txins that cause disease in humans and include bacteria, bacterial txins, viruses, fungi, rickettsiae, prtzans, and parasites. These disease-causing micrrganisms may als be referred t as infectius agents. Arthrpds and ther rganisms that transmit pathgens t animals (including humans) are called vectrs. Etilgic agents, vectrs, and materials cntaining etilgic agents are recgnized as hazardus materials. Materials cntaining etilgic agents are regularly transprted frm ne lcatin t anther by cmmn land and air carriers. Materials cntaining etilgic agents must be apprpriately packaged t prevent breakage r leakage in rder t avid expsing the package cntents t package handlers, transprters, and the general public. Materials cntaining etilgic agents must be packaged, labeled, and transprted in accrdance with all applicable regulatins. Material cntaining etilgic agents being imprted int the United States must be accmpanied by a U.S. Public Health Service imprtatin permit. Imprtatin Permits Imprtatin permits are issued nly t the imprter, wh must be lcated in the United States. The imprtatin permit, with the prper packaging and labeling, will expedite clearance f the package f infectius materials thrugh the U.S. Public Health Service Divisin f Quarantine and release by U.S. Custms. The imprter is legally respnsible fr ensuring that the freign persnnel package, label, and ship the infectius materials accrding t federal and internatinal regulatins. Shipping labels with the universal bihazard symbl, the imprter s address, the permit number, and the expiratin date are als issued t the imprter with the permit. The imprter must send the labels and ne r mre cpies f the permit t the shipper. The permit and labels infrm the U.S. Custms Service and U.S. Divisin f Quarantine persnnel f the package cntents. Federal Regulatin The imprtatin f etilgic agents is gverned by the fllwing federal regulatin: USPHS 42 CFR - Part 71 Freign Quarantine. Part Etilgic agents, hsts, and vectrs. Page 94 f 201

95 A persn may nt imprt int the United States, nr distribute after imprtatin, any etilgic agent r any arthrpd r ther animal hst r vectr f human disease, r any extic living arthrpd r ther animal capable f being a hst r vectr f human disease unless accmpanied by a permit issued by the Directr. Any imprt cming within the prvisins f this sectin will nt be released frm custdy prir t receipt by the District Directr f U.S. Custms Service f a permit issued by the Directr (Centers fr Disease Cntrl and Preventin). Items Requiring Permits Etilgic agents It is impractical t list all etilgic agents in this dcument. In general, an imprt permit is needed fr any infectius agent knwn r suspected t cause disease in humans. Bilgical materials Unsterilized specimens f human and animal tissues (such as bld, bdy discharges, fluids, excretins, r similar material) cntaining an infectius r etilgic agent require a permit in rder t be imprted. Hsts and vectrs Animals: any animal knwn r suspected f being infected with an rganism capable f causing disease in humans may require a permit issued by CDC. Imprtatin f live turtles f less than 4 inches in shell length and live nnhuman primates is regulated by the CDC s Divisin f Glbal Migratin and Quarantine ( Bats: all live bats require an imprt permit frm the CDC and the U.S. Department f Interir, Fish and Wildlife Services. The applicatin fr a CDC imprt permit fr live extic bats is at CDC Imprtatin f Animals website ( Arthrpds: any living insect r ther arthrpd that is knwn r suspected f cntaining an etilgic agent (human pathgen) requires a CDC imprt permit. Snails: snail species capable f transmitting a human pathgen require a CDC permit. Packaging Requirements Infectius materials imprted int this cuntry must be packaged t withstand breakage and leakage f cntents and be labeled, as specified in the fllwing federal regulatins: USPHS 42 CFR Part 72 - Interstate Shipment f Etilgic Agents DOT 49 CFR PART Transprtatin f Etilgic Agents Fr internatinal shipments, the Internatinal Air Transprt Assciatin s (IATA) Dangerus Gds Regulatins shuld be cnsulted. Page 95 f 201

96 Other Permits USDA and APHIS permits are required fr infectius agents f livestck and bilgical materials cntaining animal material. Tissue culture materials and suspensins f cell culture-grwn viruses r ther etilgic agents cntaining grwth stimulants f bvine r ther livestck rigins are cntrlled by the USDA because f the ptential risk f intrductin f extic animal diseases int the United States. Fr mre infrmatin, cntact USDA/APHIS at their website, Principal Investigatrs must submit USDA/APHIS permit applicatins via the IBC Office and additinal infrmatin may be fund at U.S. Fish and Wildlife Service permits are required fr certain live animals, including bats. Fr mre infrmatin, call (800) 344-WILD r visit their website, Individuals wishing t imprt select agents and txins must be registered with the CDC s Select Agent Prgram in accrdance with 42 CFR Part 73 (Pssessin, Use, and Transfer f Select Agents and Txins; Interim Final Rule) fr the select agent(s) and txin(s) listed n the imprt permit applicatin. Als, in accrdance with 42 CFR Part 73.16(a), an APHIS/CDC Frm 2 must be cmpleted and submitted t the CDC Select Agent Prgram and granted apprval prir t the shipment f the select agents r txins under the imprt permit. Additinal infrmatin can be fund at Exprtatin f Infectius Materials The exprt f a wide variety f etilgic agents f human, plant, and animal diseases may require a license frm the Department f Cmmerce. T determine if a license is necessary, visit Exprt cntrl regulatins are rather cmplex and may have multi-agency jurisdictins that must apprve activities. The Department f Cmmerce regulatins state that: Activities subject t the Exprt Administratin Regulatins (EAR) may als be cntrlled under exprt-related prgrams administered by ther agencies. Items and activities subject t the EAR are nt necessarily exempted frm the cntrl prgrams f ther agencies. Please cntact the Bisafety Prgram as sn as pssible if yu intend t exprt any bilgical materials. Page 96 f 201

97 Appendix C Labratry Ventilatin and Cntainment fr Bisafety Labratry-ventilated cntainment equipment fall int three majr categries: Labratry Chemical ( Fume ) Hds Traditinal labratry chemical (r fume) hds are designed t capture and cntrl chemical vaprs and pull them away frm the wrker. Althugh the inward flw f air prtects the user, chemical hds d nt prtect the prduct (the desired rganism being manipulated). Hrizntal Laminar Flw Clean Bench With hrizntal laminar flw clean benches, HEPA-filtered air flws hrizntally acrss the wrkspace directly tward the user. These clean benches prvide prduct prtectin and were riginally designed t prvide a particulate-free envirnment fr the manufacture f semicnductr cmpnents. Clean benches prvide prduct prtectin against micrbial cntaminatin, but they d nt prvide persnal r envirnmental prtectin. In fact, the hrizntal flw f air will blw bilgical agents directly tward the user and int the labratry. Clean benches are nt a bilgical safety cabinet, and they shuld nt be used with any materials (bilgical, chemical, r radilgical) requiring cntainment fr prtectin f persnnel r the envirnment. Clean benches are acceptable fr tissue culture wrk nly with cell lines cnsidered t represent lw risk (BSL-1 agents) t labratry wrkers (including immuncmprmised individuals wh may frequent the lab). Human cell lines and nnhuman primate cell lines are generally cnsidered t be BSL-2 agents and wuld nt be suitable fr use in a clean bench. Bilgical Safety Cabinets Bilgical safety cabinets (BSCs) are divided int Class I, II, and III (see schematic belw). Class II BSCs are subdivided int type A and type B. All BSCs prvide persnnel and envirnmental prtectin, with Class II BSCs als prviding prduct prtectin. Persnnel prtectin is achieved by inward airflw thrugh the frnt f the cabinet. Prduct prtectin is achieved by dwnward HEPA-filtered airflw frm the tp f the cabinet. Envirnmental prtectin is achieved by HEPA filtratin f exhaust air. Page 97 f 201

98 New NSF Classificatin, Adpted 2002 A1 Previus NSF Classificatin Class II, Type A General Descriptin 70% air recirculated; 30% exhausted frm a cmmn plenum t the rm; 75FPM intake; may have bilgically cntaminated psitive pressure plenum A2 A2 Class II, Type A/B3 Class II, Type B3 70% air recirculated; 30% exhausted frm a cmmn plenum t the rm; 100FPM intake; bilgically cntaminated plenum under negative pressure r surrunded by negative pressure 70% air recirculated; 30% exhausted frm a cmmn plenum t a facility exhaust system; 100FPM intake; bilgically cntaminated plenum under negative pressure r surrunded by negative pressure B1 Class II, Type B1 40% air recirculated; 60% exhausted frm cabinet; exhaust air pulled thrugh dedicated exhaust duct int facility exhaust system; 100FPM intake all bilgically cntaminated plenums are negative t the rm r surrunded by negative pressure plenums Page 98 f 201

99 B2 Class II, Type B2 0% air recirculated; 100% exhausted frm cabinet exhaust air pulled thrugh dedicated exhaust duct int facility exhaust system; 100FPM intake all ducts and plenums are under negative pressure all cntaminated ducts are under negative pressure r surrunded by directly exhausted negative pressure ducts r plenums Certificatin f BSCs Generally, cmmercial BSCs are tested by the cabinet manufacturer in accrdance with Natinal Sanitatin Fundatin (NSF) criteria. Cabinets that meet the NSF criteria fr perfrmance characteristics, including bilgical cntainment, ventilatin, cabinet leakage, and HEPA filter leakage, are NSF certified. Field certificatin f BSCs is als required t ensure that the cabinet still perfrms as it did when it btained NSF certificatin at the factry. NIH requires field certificatin under the fllwing circumstances: (1) upn installatin f a new BSC; (2) annually thereafter; (3) after repair r maintenance is perfrmed; and (4) after the BSC is relcated. CDC recmmends that BSCs be recertified annually t ensure fr prper functin. They will als be recertified after being mved t ensure that they have nt been damage. Labratries are respnsible fr ensuring that the BSCs are recertified in a timely manner. Labratries at CRC will cntact EHS at (617) Labratries at BUMC will cntact the certificatin cntractr directly. The cntact infrmatin t reach the cntractr is indicated n the certificatin sticker affixed n the frnt f the BSC. NSF standard 49 prvides criteria fr cnstructin f BSCs, testing by manufacturers (including bilgical cntainment testing), and field certificatin. NSF has als established a certificatin prgram fr field certifiers t ensure a minimum level f cmpetency and prfessinalism. It is recmmended that NSF field certifiers be used fr field certificatin f BSCs. Field certificatin tests include: Primary tests (BSC perfrmance): a. Inflw test b. Dwn-flw test c. Smke pattern test Page 99 f 201

100 d. HEPA filter leakage e. Cabinet leakage (when BSC is newly installed, relcated, r maintenance has been perfrmed that invlved remval f access panels) Additinal tests (wrker cmfrt and safety), perfrmed at discretin f certifier: a. Nise b. Vibratin c. Lighting d. Electrical leakage, plarity, and grund circuit resistance Hrizntal Flw Clean Bench A. frnt pening B. supply grille C. supply HEPA filter D. supply plenum E. blwer F. grille Class I BSC A. frnt pening B. sash C. exhaust HEPA D. exhaust plenum Page 100 f 201

101 Class II Type A BSC A. frnt pening B. sash C. exhaust HEPA filter D. rear plenum E. supply HEPA filter F. blwer Class II Type B3 BSC (Tabletp mdel) A. frnt pening B. sash C. exhaust HEPA filter D. supply HEPA filter E. psitive pressure plenum F. negative pressure plenum Page 101 f 201

102 Class II Type B1 BSC A. frnt pening B. sash C. exhaust HEPA filter D. supply HEPA filter E. negative pressure exhaust plenum F. blwer G. additinal HEPA filter fr air supply Class II Type B2 BSC A. frnt pening B. sash C. exhaust HEPA filter D. supply HEPA filter E. negative pressure exhaust plenum F. supply blwer G. filter screen Page 102 f 201

103 Appendix D Autclave Quality Assurance Prgram Autclaving is an accepted prcedure fr the decntaminatin f certain bihazardus waste. Bilgical cultures and stcks, cntaminated slid waste, and liquid waste can be sterilized thrugh autclaving. After sterilizatin in a steam autclave, these materials are cnsidered nn-infectius. At BU/BMC, all autclaved waste is placed int the slid bihazard waste stream. Materials that cntain hazardus chemicals r radiistpes are nt t be autclaved. T ensure that bihazardus waste is prperly decntaminated during autclaving, the fllwing prcedures shuld be fllwed by labratry persnnel 1. Infectius waste must be treated in an autclave fr a minimum f 30 minutes at 121 C (250 F); hwever, the ttal prcessing time required t decntaminate infectius waste depends n the specific lading factrs (cntainer type, water cntent, quantity, etc.). A ttal prcessing time f 60 minutes is recmmended fr gravity displacement autclaves and 10 minutes fr vacuum-type autclaves (132 C). Sterilizatin by autclaving is accmplished thrugh expsure and penetratin f the cntaminated material by superheated steam fr an adequate amunt f time. Because steam will nt penetrate a sealed plastic autclave bag, bags cntaining dry lads must nt be tightly sealed (rubber band clsures will allw bags t breathe ) r adequate amunts f water must be added t the lad. Cnsult the manufacturer s instructins fr sterilizing materials inside plastic autclave bags. Liquid waste may als be autclaved in lieu f adding an apprpriate chemicals disinfectant, and dispsed in the sink. Animal carcasses frm the ABSL3 and ABLS4 will be autclaved inside an autclavable bag. 2. All autclaved waste must include a steam sterilizatin indicatr (the use f bihazard bags with a built-in indicatr is recmmended). 3. Steam autclaves used t treat infectius waste must perate at a minimum temperature f 121 C. The perating temperature f the autclave must be verified fr each run by maintaining a recrd f the temperature either as a chart r paper tape recrding r a manual recrding in a lgbk. 4. On a mnthly basis, cnfirm that adequate sterilizatin cnditins are being met thrugh the use f ampules cntaining heat-resistant spres (Gebacillus stearthermphilus) placed in the center f an autclave lad. In cnjunctin with the B. stearthermphilus testing, measure and recrd the maximum temperature achieved during the autclave cycle thrugh the use f a maximum registering (r hlding ) thermmeter r calibrated data lgger fr full cycle. 5. Maintain recrds f B. stearthermphilus testing and maximum autclave temperature recrdings fr a minimum f ne year (see Autclave QC Lg at end f appendix). Page 103 f 201

104 Mnthly Spre Testing Prcedure 1. Place ampule f B. stearthermphilus spres and hlding thermmeter r data lgger in the center f an autclave lad. 2. Prcess the lad under nrmal perating prcedures. 3. The highest temperature indicated n the hlding thermmeter is entered n the Autclave QC Lg. If this temperature is less than 121 C, the autclave is nt t be used t treat infectius waste until it has been repaired and passes retesting. In the interim, tag the autclave as Nt Apprved fr Infectius Waste. 4. Incubate the autclaved ampule and a nn-autclaved, cntrl ampule accrding t the manufacturer s instructins (nrmally C fr 24 t 48 hurs). 5. If a clr change ccurs, the sterilizatin prcess was unsuccessful. Discntinue use f the autclave until it is repaired and passes retesting. Tag the autclave as Nt Apprved fr Infectius Waste until the autclave passes retesting. 6. Indicate test results n Autclave QC Lg (see end f appendix) and retain fr at least ne year. Page 104 f 201

105 AUTOCLAVE QC LOG Year: ; Autclave Lcatin: Manufacturer: ; Mdel: Serial Number: Autclave Testing Instructins: 1. Perfrm autclave QC tests mnthly. 2. Place B. stearthermphilus spre ampule and hlding thermmeter in the center f an autclave lad. 3. Prcess the lad under nrmal perating prcedures. 4. Recrd the highest temperature indicated n the hlding thermmeter. If this temperature is less than 121 C, the autclave is nt t be used t treat infectius waste until it has been repaired and passes retesting. 5. Incubate the autclaved ampule and a nn-autclaved cntrl ampule accrding t the manufacturer s instructins (nrmally C fr 24 t 48 hurs). 6. If a clr change ccurs, the sterilizatin prcess was unsuccessful. Discntinue use f the autclave (fr infectius waste) until it is repaired and passes retesting. 7. Recrd testing results n Autclave QC Lg and retain fr at least ne year. Jan. Feb. Mar. Apr. May June July Aug. Sept. Oct. Nv. Dec. Date Operatr Cycle Time Cycle Temp Results Cmments Page 105 f 201

106 Appendix E Bisafety Level 2 (BSL-2) Requirements Bisafety Level 2 (BSL-2) is suitable fr experiments invlving agents f mderate ptential hazard t persnnel and the envirnment. Fr example: Micrrganisms f lw bihazard ptential, such as thse in Risk Grup 2 r BSL-2. Recmbinant DNA activity requiring BSL-2 physical cntainment including animal studies that invlve the cnstructin f transgenic animals. Nn-recmbinant cell and/r tissue culture systems that require this level f cntainment. Oncgenic viral systems classified as lw risk. Prductin activities with Risk Grup 1 rganisms. The cntrl f ptential bihazards at the BSL-2 level is prvided by use f standard micrbilgical practices with the additin f persnnel prtective equipment (lab cat and glves). The fllwing are prcedures are used with BSL-2 cntainment requirements. They are based n the recmmendatin f the Bisafety in Micrbilgical and Bimedical Labratries (BMBL) 5 th Editin, Standard Micrbilgical Practices Access t the labratry is limited r restricted at the discretin f the labratry directr when experiments are in prgress. Persns wash their hands after they handle viable materials, after remving glves, and befre leaving the labratry. Eating, drinking, smking, handling cntact lenses, and applying csmetics are nt permitted in the wrk areas. Fd is stred utside the wrk area in cabinets r refrigeratrs designated fr this purpse nly. Persnnel wh use cntact lenses will cnsult with EHS if required t use eye prtectin in the lab. Muth pipetting is prhibited; mechanical pipetting devices are used. Plicies fr the safe handling f sharps are instituted. All prcedures are perfrmed carefully t minimize the creatin f splashes r aersls. Wrk surfaces are decntaminated upn cmpletin f wrk, r at the end f the day, and after any spill Page 106 f 201

107 r splash f viable material with disinfectants that are effective against the agents f cncern. All cultures, stcks, and ther regulated wastes are decntaminated befre dispsal by an apprved decntaminatin methd, such as autclaving. Materials t be decntaminated utside the immediate labratry are placed in a durable, leak-prf cntainer and clsed fr transprt frm the labratry. Materials t be decntaminated ff-site are packaged in accrdance with applicable lcal, state, and federal regulatins befre remval frm the facility. An insect and rdent cntrl prgram is in effect. Special Practices Access t the labratry is limited r restricted by the labratry directr when wrk with infectius agents is in prgress. In general, persns wh are at increased risk f acquiring infectin, r fr whm infectin may have serius cnsequences, are nt allwed in the labratry r animal rms. Fr example, persns wh are immuncmprmised r immunsuppressed may be at increased risk f acquiring infectins. The labratry directr in cnsultatin with Research Occupatinal Health Prgram has the final respnsibility fr assessing each circumstance and determining wh may enter r wrk in the labratry r animal rm. The Principal Investigatr r Labratry Directr establishes plicies and prcedures whereby nly persns wh have been advised f the ptential hazards and meet specific entry requirements (e.g., immunizatin) may enter the labratry. A bihazard sign must be psted n the entrance t the labratry when etilgic agents are in use. Apprpriate infrmatin t be psted includes the agent(s) in use; the bisafety level; the required immunizatins; the investigatr s name and telephne number; any persnal prtective equipment that must be wrn in the labratry; and any prcedures required fr exiting the labratry. Labratry persnnel receive apprpriate immunizatins r tests fr the agents handled r ptentially present in the labratry (e.g., hepatitis B vaccine r TB skin testing). Bisafety prcedures are incrprated int standard perating prcedures r in a bisafety manual adpted r prepared specifically fr the labratry by the labratry directr. Persnnel are advised f special hazards and are required t read and fllw instructins n practices and prcedures. The Principal Investigatr r Labratry Directr ensures that labratry and supprt persnnel receive apprpriate training abut the ptential hazards assciated with the wrk invlved; the necessary precautins t prevent expsures; and the expsure evaluatin prcedures. Persnnel receive annual updates r additinal training as necessary fr prcedural r plicy changes. A high degree f precautin must always be taken with any cntaminated sharp items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels. Needles and syringes r ther sharp instruments shuld be restricted in the labratry fr use nly when there is n alternative, such as parenteral injectin, phlebtmy, r aspiratin f fluids frm labratry animals and diaphragm bttles. Plasticware shuld be substituted fr glassware whenever pssible. Page 107 f 201

108 Only needle-lcking syringes r dispsable syringe-needle units (i.e., needle is integral t the syringe) are used fr injectin r aspiratin f infectius materials. Used dispsable needles must nt be bent, sheared, brken, recapped, remved frm dispsable syringes, r therwise manipulated by hand befre dispsal; rather, they must be carefully placed in cnveniently lcated puncture-resistant cntainers used fr sharps dispsal. Nn-dispsable sharps must be placed in a hard-walled cntainer fr transprt t a prcessing area fr decntaminatin, preferably by autclaving. Syringes that re-sheathe the needle, needleless systems, and ther safety devices are used when apprpriate. Brken glassware must nt be handled directly by hand, but must be remved by mechanical means such as a brush and dustpan, tngs, r frceps. Cntainers f cntaminated needles, sharp equipment, and brken glass are decntaminated befre dispsal accrding t any lcal, state, r federal regulatins. Cultures, tissues, specimens f bdy fluids, r ptentially infectius wastes are placed in a cntainer with a cver that prevents leakage during cllectin, handling, prcessing, strage, transprt, r shipping. Labratry equipment and wrk surfaces shuld be decntaminated with an effective disinfectant n a rutine basis; after wrk with infectius materials is finished; and especially after vert spills, splashes, r ther cntaminatin by infectius materials. Prir t its remval frm the facility, cntaminated equipment must be decntaminated accrding t any lcal, state, r federal regulatins befre it is sent fr repair r maintenance r packaged fr transprt in accrdance with applicable lcal, state, r federal regulatins. Spills and accidents that result in vert expsures t infectius materials are immediately reprted t the Principal Investigatr and Labratry Directr. Medical evaluatin, surveillance, and treatment are prvided as apprpriate, and written recrds are maintained. Sinks in the BSL-2 area shuld be cleared rutinely using apprpriate disinfectant such as a chlrinecntaining abrasive and flushed with a suitable chemical decntaminant. Water baths and all water reservirs shuld be washed peridically with a suitable chemical decntaminant. Once a mnth, wrk spaces that d nt get daily attentin with germicide shuld be cleaned, as well as ther lab areas where clutter accumulates (e.g., strage areas). The labratry will set up a rutine schedule t perfrm surface cleaning with apprpriate chemical disinfectant f large equipment (such as incubatrs) as part f labratry gd practices. Supplies shuld be rtated and utdated material thrwn ut. Unlabeled material shuld be eliminated. Clutter shuld be cleaned up. Custdial services: nly persnnel with apprpriate authrizatin may enter a BSL-2 facility while BSL-2 research activity is in prgress. Animals nt invlved in the wrk being perfrmed are nt permitted in the lab. Page 108 f 201

109 Safety Equipment (Primary Barriers) Prperly maintained bilgical safety cabinets, preferably Class II, r ther apprpriate persnal prtective equipment r physical cntainment devices are t be used when: Prcedures that have the ptential t create infectius aersls r splashes are cnducted. These may include centrifuging, grinding, blending, vigrus shaking r mixing, snic disruptin, pening cntainers f infectius materials whse internal pressures may be different frm ambient pressures, inculating animals intranasally, and harvesting infected tissues frm animals r embrynate eggs. High cncentratins r large vlumes f infectius agents are used. Such materials may be centrifuged in the pen labratry if sealed rtr heads r centrifuge safety cups are used, and if these rtrs r safety cups are pened nly in a bilgical safety cabinet. Face prtectin (gggles, mask, face shield, r ther splatter guard) is used fr anticipated splashes r sprays f infectius r ther hazardus materials t the face when the micrrganisms must be manipulated utside the BSC. Prtective labratry cats, gwns, smcks, r unifrms designated fr lab use are wrn while in the labratry. This prtective clthing is remved and left in the labratry befre leaving fr nn-labratry areas (e.g., cafeteria, library, administrative ffices). All prtective clthing is either dispsed f in the labratry r laundered by the institutin; it shuld never be taken hme by persnnel. Glves are wrn when hands may cntact ptentially infectius materials, cntaminated surfaces, r equipment. Wearing tw pairs f glves may be apprpriate. Glves are dispsed f when vertly cntaminated and remved when wrk with infectius materials is cmpleted r when the integrity f the glve is cmprmised. Dispsable glves are nt washed, reused, r used fr tuching clean surfaces (keybards, telephnes, etc.). They shuld nt be wrn utside the lab. Alternatives t pwdered latex glves shuld be available. Hands are washed fllwing remval f glves. Prcedures fr Receiving and Inspecting Samples The PI will designate a respnsible persn fr the purchase f all infectius materials t be used in the BSL-2 lab. Infectius materials will be shipped t the labratry in accrdance with the apprpriate Department f Transprtatin (DOT) and the Internatinal Air Transprtatin Assciatin (IATA) standards fr shipping f infectius bilgical materials. Upn receipt f the package, it will be placed n a tray cvered with absrbent material and pened in the Bilgical Safety Cabinet prevent any ptential expsure t persnnel in case the cntainer leaked during transprt. Persnnel assigned t pen packages will wear lab smck, glves, and eye prtectin. If any cntainers are fund t be damaged, leaking r therwise cntaminated, they will be immediately Page 109 f 201

110 islated int a plastic bag alng with all packaging materials. The spill will be disinfected and clean up. The Principal Investigatr, lab directr r designee will be ntified immediately. The incident will reprted t EHS and as necessary, t apprpriate agencies. If, after inspectin, the samples are intact, they can be placed int labeled secndary cntainers (unbreakable plastic cntainers r metal tubes) and then transferred t a strage area. Only staff wh are authrized t d s can remve samples frm strage. Remval and use f all such materials must be entered int the lgbk. Unused cultures can be returned t strage after the uter cntainer has been prperly disinfected. Labratry Facilities (Secndary Barriers) In a BSL-2 lab, the fllwing cnditins are t exist: Lckable drs shuld be prvided fr facilities that huse restricted areas. Cnsideratin shuld be given t lcating new labratries away frm public areas. Each labratry cntains a sink fr handwashing. The labratry is designed s that it can be easily cleaned. Carpets and rugs in labratries are inapprpriate. Bench tps are impervius t water and resistant t mderate heat and the rganic slvents, acids, alkalis, and chemicals used t decntaminate the wrk surfaces and equipment. Labratry furniture is capable f supprting anticipated lading and uses. Spaces between benches, cabinets, and equipment are accessible fr cleaning. Chairs and ther furniture used in labratry wrk shuld be cvered with a nn-fabric material that can be easily decntaminated. Bilgical safety cabinets shuld be installed in such a manner that fluctuatins f the rm s air supply and exhaust air d nt cause them t perate utside their parameters fr cntainment. Lcate BSCs away frm drs, windws that can be pened, heavily traveled labratry areas, and ther ptentially disruptive equipment s as t maintain the BSC s air flw parameters fr cntainment. An eyewash statin is readily available. Illuminatin is adequate fr all activities, aviding reflectins and glare that culd impede visin. There are n specific ventilatin requirements. Hwever, planning f new facilities shuld cnsider mechanical ventilatin systems that prvide an inward flw f air withut recirculatin t spaces utside f the labratry. If the labratry has windws that pen t the exterir, they are fitted with fly screens. Page 110 f 201

111 Appendix F Bisafety Level 3 (BSL-3) Requirements and Practices Bisafety Level 3 (BSL-3) is the recmmended cntainment fr wrk with agents r txins that may cause serius r ptentially fatal disease thrugh inhalatin expsure. Wrk at BSL-3 requires enhanced facility design, peratinal cntrls and special practices which will be utlined in this sectin. BSL-3 facilities cnduct research invlving bilgical materials f risk grup 3 (r high titers f RG2) and are designed and verified with facility and ventilatin features t accmmdate the safe handling and cntainment f such agents. The architectural and engineering plans fr all BSL-3 facilities will be reviewed and apprved in advance by EHS t ensure that they in cmpliance with the CDC/NIH s Bisafety in Micrbilgical and Bimedical Labratries (BMBL current editin) requirements. Facility specific standard perating prcedures (SOP s) are reviewed by EHS and research prtcls are reviewed and apprved by the BU/BUMC Institutinal Bisafety Cmmittee t determine adequacy fr the use f the prpsed bilgical agents. Additinally, all wrk with BSL-3 and abve agents must be registered with the Bstn Public Health Cmmissin. The fllwing utlines the general requirements fr BSL-3 facilities at Bstn University. Special requirements dealing with Select Agents and Select Agent facility dcumentatin is cvered in the facility specific Labratry Manuals. Administratin Rles and Respnsibilities The Principal Investigatr (PI) r Labratry Directr is respnsible fr: Ensuring that persnnel are adequately trained Ensuring that individuals wrking in the facility are experienced and prficient in handling the bilgical agents at the apprpriate level f cntainment Ensuring that any visitr r cntractr is escrted by an individual trained and apprved t enter the facility Ensuring that the BSL-3 facility and the practices within the facility are in cmpliance with all federal, state, and lcal regulatins The Labratry Manager is respnsible fr: Overseeing the day t day peratins f the BSL-3 facility Ensuring that safety practices are adhered t in the facility Cntacting the PI and EHS in the event f an emergency within the facility Cntacting Facilities and EHS in the event f a facility perfrmance issue Ensuring that the BSL-3 facility and the practices within the facility are in cmpliance with all Page 111 f 201

112 federal, state, and lcal regulatins EHS is respnsible t: Cnduct quarterly inspectin f the BSL-3 facility Prvide the necessary safety training t individuals that will enter the facility Respnd t any emergency situatin in the facility Ensure that the BSL-3 facility and the practices within the facility are in cmpliance with all federal, state, and lcal regulatins ROHP is respnsible fr: Prviding medical evaluatins and screening fr persnnel wh will wrk in the facility Prviding medical surveillance prgram fr all persnnel wrking in the facility BU Facilities Grup is respnsible fr: Maintaining the BSL-3 facility in cmpliance with federal, lcal, and state regulatins Review and bserve the facility re-certificatin prcess Prviding a facility preventative maintenance prgram Prviding the facility with a pest cntrl prgram The facilities manager will supervise preventative maintenance BSL-3 Standard Micrbilgical Practices Access t BSL-3 labs is restricted t thse individuals apprved fr entry by the labratry directr in cnjunctin with EHS and Public Safety. Entry t cntainment area is thrugh a cntrlled access anterm. Nte: Access t Select Agent BSL-3 labs is utlined in the facility security plan and facility specific SOP s. The lab directr establishes plicies and prcedures whereby nly persns wh have been advised f the ptential hazard, wh meet any specific training and entry requirements, and wh cmply with all prcedures may enter the wrk area(s). Persnnel entering the facility are advised f specific hazards and are required t read and fllw instructins n practices and prcedures. Cautin must be used with any sharp items. Use f hypdermic needles and Pasteur pipettes is restricted in the BSL-3 lab. If needles are t be used, safe r prtected needle devices are recmmended. Extreme care shuld be used t avid aut-inculatin and aersl generatin. Cntaminated sharps must be prmptly placed in a puncture-resistant sharps cntainer and Page 112 f 201

113 decntaminated befre dispsal. Brken glassware must nt be handled directly by hand. Plastic ware shuld be substituted whenever pssible. Persns must wash their hands befre exiting the facility. Eating, drinking, smking, applying csmetics, inserting r remving f cntact lenses, and string fd are prhibited in the lab. Muth pipetting is prhibited. All prcedures with infectius materials are perfrmed in a bilgical safety cabinet r ther physical cntainment device t minimize the expsure f persnnel t aersls. Wrk surfaces plus cunters and sinks are decntaminated at least nce every wrk shift r prir t use by ther lab persnnel and immediately after any spill f viable material. Agent apprpriate disinfectants are maintained in the labratry. Labratry equipment and wrk surfaces are t be decntaminated using an agent apprpriate disinfectant after wrk with infectius materials, especially after spills r splashes. Cntaminated equipment shuld be decntaminated befre being sent fr repair, maintenance, r transprt, and befre remval frm the facility. All infectius wastes are decntaminated befre dispsal by apprved methds. Nn dispsable items are decntaminated befre washing and reuse. An insect and rdent cntrl prgram is in effect and is maintained the labratry peratins r the Facilities grup. Lwer cntainment experiments in the same lab must fllw BSL-3 practices. BSL-3 Special Practices The lab directr establishes plicies and prcedures whereby nly persns wh have been advised f the ptential hazard, wh meet any specific training and entry requirements, and wh cmply with all prcedures may enter the wrk area(s). Lab drs are kept clsed while wrk is in prgress. Apprpriate BSL-3 signage is psted n the utside f the BSL-3 suite alng with a universal bihazard warning sign and emergency cntact infrmatin. The sign identifies the agent, lists the name and telephne number f the lab directr r ther respnsible persn(s), and indicates any special requirements fr entering the lab (e.g., immunizatin, persnal prtective measures). Equipment used t stre, grw r prcess cultures shall als be psted with the universal bihazard label. The lab directr cntrls access t the lab and restricts it t prperly trained persns wh are advised f the risks and whse presence is required fr the wrk being cnducted. Persns wh are at an increased risk f acquiring infectin r fr whm infectin may be unusually hazardus are nt allwed in the wrk area. Minrs are nt allwed in the facility. N custdial r ther maintenance Page 113 f 201

114 persnnel are permitted t enter the BSL-3 lab unless authrized t d s and escrted by authrized persnnel. The labratry directr has the final respnsibility fr assessing each circumstance. Labratry persnnel are required t be medically cleared by the Research Occupatinal Health Prgram prir t wrking in a BSL-3 labratry (See Appendix R Medical Surveillance Prgram) where apprpriate testing and immunizatins will be perfrmed based n ccupatinal expsure and agents handled. The lab directr ensures that lab persnnel receive apprpriate training n the wrk s ptential hazards, the necessary precautins t prevent expsure, and emergency prcedures. Persnnel receive additinal training as needed. All labratry persnnel must demnstrate prficiency in the handling f hazardus agents/materials and in ther practices specific t the facility befre wrking with infectius materials. Subject matter experts, wh are experienced in wrking with these agents/materials help prvide the training and supervisin. N wrk with infectius materials is cnducted in pen vessels n the pen bench. All activities invlving such materials are perfrmed in BSCs r ther physical cntainment devices within the cntainment mdule. When a prcedure cannt be perfrmed in a BSC, a cmbinatin f PPE and ther cntainment devices must be used. Opening centrifuge rtr heads and caps must als be dne inside a BSC. All ptentially cntaminated items frm the wrk area(s) are sterilized by a methd shwn t be effective fr the rganism in use befre dispsal r reuse. Labratry persnnel must be prperly trained and equipped t decntaminate and clean up spills f infectius materials. See Chapter 8. Spills and accidents that result in vert r ptential expsure t infectius material are immediately reprted t the lab directr, wh then ntifies EHS. Medical evaluatin, surveillance, and treatment are prvided as apprpriate, and written recrds are maintained. Animals and plants nt part f the wrk being cnducted are nt permitted in a BSL-3 lab. Safety Equipment (Primary Cntainment Barriers and PPE) Bilgical safety cabinets are used fr all pen cntainer manipulatins f infectius materials. In additin t a BSC, ther apprpriate cmbinatins f persnal prtective equipment (such as special clthing r face prtectin) and physical cntainment devices (such as centrifuge safety caps and sealed centrifuge rtrs) are used fr all activities that pse a threat f aersl expsure t infective agent(s).this special equipment must be used fr manipulatins f cultures and infected animals. Use f respiratry prtectin r enhanced envirnmental prtectin will be based n risk assessment. Persnnel whse jbs require the use f a respiratr must cmplete an OSHA Respiratr Questinnaire Page 114 f 201

115 fr medical clearance by the Research Occupatinal Health Prgram n an annual basis and be fit tested and instructed by the Industrial Hygiene Office. In the lab, dispsable PPE that prtects street clthing (e.g., lng-sleeved, slid frnt r wrap-arund gwns, scrub suits, r cveralls) is required. When wrking with BSL-3 infectius agents, gwns shuld be impervius t liquids and have elasticized cuffs. Lab clthing is nt wrn utside the lab and is decntaminated befre dispsal. Frnt-buttn lab cats are nt suitable fr a BSL-3 lab. Street clthing, such as cats and hats, are nt kept in the lab. Duble glves are wrn when wrking with infectius materials r when hands may cme in cntact with cntaminated surfaces r equipment. Glves must be changed when cntaminated; glve integrity is cmprmised r therwise apprpriate. Dispsable glves must never be washed r reused. Cntaminated glves must be dispsed f in cntaminated labratry waste. When wrk with cntaminated materials is cmpleted, glves must be remved and hands washed befre exiting the facility. Labratry Facility (Secndary barriers) The labratry is segregated frm areas that are pen t unrestricted traffic in the building. Access int the lab is thrugh at least tw sets f self-clsing drs. A changing rm may be included in the passageway between the self-clsing drs. The lab cntains a sink fr hand-washing that is ft perated and is lcated near the exit dr. Handwashing facilities with autmatic faucets are als lcated in the anterm. The facility must be designed s that it is easily cleanable. Carpets and rugs are nt permitted. Seams, walls, flrs and ceiling surfaces shuld be sealed. Spaces arund ventilatin penings, light fixtures, wall plates, and any wall penetratins shuld be sealed. Flrs must be slip resistant, impervius t liquids, and resistant t chemicals. Walls and ceilings shuld be cnstructed t be sealed with a smth finish and shuld be easily cleanable. Decntaminatin f the entire labratry shuld be cnsidered when there has been a grss cntaminatin, significant changes t labratry usage, fr majr renvatins r fr maintenance shutdwns. The interir surfaces f walls, flrs, and ceilings are water resistant. Penetratins in these surfaces are sealed. Bench tps are impervius t water and resistant t acids, alkalis, rganic slvents, and mderate heat. Spaces between benches, cabinets, and equipment are accessible fr cleaning. Windws must be sealed. Page 115 f 201

116 An autclave fr decntaminatin f labratry waste is lcated in the labratry. The lab is under negative pressure and has directinal airflw, with air drawn in thrugh the clean entry area. The exhaust air is nt recirculated t any area and is HEPA filtered prir t being discharged t the utside. The exhaust is dispersed away frm ccupied areas and air intakes. Persnnel must verify that the directin f airflw int the lab is crrect befre entering. Audible alarms will ntify labratry persnnel if labratry airflw is disrupted. Labratry exhaust must nt re-circulate t any ther areas f the building. HEPA husings shuld have gas-tight islatin dampers, decntaminatin prts, and/r bag in/bag ut capability. HEPA filter husing shuld allw fr leak testing f each filter and assembly. Filters and husing shuld be certified at least annually. Bisafety cabinets shuld be certified annually t assure perfrmance. Equipment that may prduce infectius aersls must be cntained by primary barrier devices that exhaust air thrugh HEPA filtratin r equivalent technlgy prir t discharge. Vacuum lines are prtected by in line HEPA filters and disinfected traps. An eyewash and/r safety shwers is readily available where applicable. Facilities must be re-verified annually. Additinal design features are listed in Appendix C. Remval f Materials frm the Facility Material t be remved frm the facility shall be prperly decntaminated by autclaving r by chemical disinfectin. Autclaving is the preferred mde f decntaminatin and shuld be used fr all refuse and lab-ware. Items which cannt be autclaved shall be decntaminated by wiping the surfaces thrughly with an agent apprpriate chemical disinfectant. Nte: The Massachusetts Water Resurces Authrity prhibits dispsal f mercury-cntaining liquids int waste water. BU/BMC recmmends the use f bleach, which is mercury free. Befre remving any majr piece f equipment frm the BSL-3 lab, EHS must be ntified t determine the best curse f actin. Depending n the specific situatin, the facility may be restricted r shut dwn during remval f the equipment. Reusable cntainers include tissue culture media bttles, reagent bttles, cylinders, and beakers. They shall be decntaminated by immersin in an apprpriate disinfectant such as 10% bleach slutin fr at least 10 minutes (care shuld be taken t fill cmpletely). The items will be drained and washed and autclaved ut f the facility. Autclaved reusable cntainers will be kept t a minimum. In case f a breakdwn in the -70 C freezer, frzen material will be place int secndary shatterprf transprt cntainers, and remved either t a freezer inside anther cntainment lab r, if space is nt available, t a labeled backup freezer in the institute. If remved frm the facility t a back-up freezer, the utside f the strage and transprt cntainers will be disinfected with agent apprpriate disinfectant. Page 116 f 201

117 DNA, RNA, prteins, r ther bilgical materials prepared frm infectius agents, r infected cells created by using extractin prcedures that cmpletely inactivate the agent (as apprved by IBC), may be remved frm the cntainment labratry. Such materials must be cnfirmed t cntain n infectius BSL-3 materials prir t remval. If materials are remved frm the facility, the utside f the strage and transprt cntainers will be disinfected with agent apprpriate disinfectant. Any infectius DNA, virus, r cellular material that is remved frm the facility will be recrded with the date, the type f material, the amunt f material, and the rigin r destinatin f the material. Shipping f materials frm the BSL-3 shall be packaged accrding t Department f Transprtatin requirements. The primary cntainer will be decntaminated with agent-apprpriate disinfectant befre remval frm the BSC and then placed inside a clean secndary cntainer utside the cabinet. Glves will be changed between each step f packaging. Absrbent material will be placed between the primary and secndary cntainers. The utside f the secndary cntainer will then be decntaminated with agent-apprpriate disinfectant prir t remval frm the facility. EHS must be cntacted in advance f shipping such materials and shuld be cntacted prir t preparing the samples. Cleaning the Facility All persnnel will dn apprpriate persnal prtective equipment prir t starting wrk in/ r cleaning the facility. Researchers will perfrm all daily husekeeping rutines within the BSL-3 lab, including trash remval. The general BUMC husekeeping staff will nt enter the labratry t d any regular cleaning. All the cleaning and decntaminatin prcedures shall be perfrmed nly by individuals authrized t wrk in the BSL-3 facility. Wrk surfaces are decntaminated when wrk is finished, at the end f every wrk day, and immediately after any spill f viable material. Large equipment, such as incubatrs and centrifuges, will have inner and uter surfaces damp-wiped with disinfectant n a rutine basis. Sinks in the BSL-3 lab shuld be cleaned and flushed with agent apprpriate disinfectants. Water baths and all water reservirs shall be rinsed peridically with a suitable chemical decntaminant if cpper sulfate (r ther antimicrbial) has nt been added t the water. On a rutine basis, the flr f the BSL-3 cntainment is thrughly wiped dwn with a suitable chemical decntaminant and wrk spaces that d nt get daily attentin are cleaned up with germicide. Slid waste is decntaminated by autclaving prir t remval frm the facility, and dispsed f by authrized persnnel. Entry/Exit practices Lab ntebks are nt t be brught int the cntainment sectin f the lab; if they are, they may nt be remved. Page 117 f 201

118 The lab is als designed fr cmputer capability and faxing. Other ptins include data cllectin with printed utput elsewhere; digital writing tablets; r audi tape recrds. D nt enter the lab unless air flws int the lab when pening the dr. Immediately reprt prblems t the Medical Campus Cntrl Center at (617) r EHS at (617) Check that phne is peratinal and emergency numbers are psted. D nt wrk in the suite unless a means t summn help in an emergency is available. Any prblems shuld be reprted t Maintenance. Befre beginning wrk, check the cnditin f wrk areas. Areas shuld be clean. If they aren t, n assumptins shuld be made abut what materials were left. The last persn lgged in shuld be called fr assistance. Wipe dwn wrk areas with disinfectant befre beginning wrk. All wrk with pen cntainers f ptentially infectius material is cnfined t bilgical safety cabinets. Keep grills clear and the sash at a prper level. Use minimum equipment in the cabinet t allw fr effective air flw. Wrkflw shuld be frm clean t dirty in the Bisafety Cabinet. Disinfect r remve yur uter glves prir t remving yur hands frm the bisafety cabinet. Wipe dwn the wrk area with disinfectant after cmpleting wrk. Discard uter glves and any cntaminated clthing in the labratry waste. Remve carefully t keep the exterir f all cntaminated items away frm the skin. Labratry Layut The labratry s layut, the equipment present, and ther physical features are dependent n the activities cnducted within the facility. The labratry is laid ut in a manner that the least-hazardus activities are clsest t the entrance/exits and the flw f persnnel is in a directin that reduces the ptential fr spread f cntaminatin within the labratry. EHS will review all facility layuts with the labratry directr t ensure that the final cnfiguratin and placement f equipment meets the labratry s functinal and safety needs. The fllwing guidelines are intended t prvide the labratries with cmmnly acceptable criteria. All BSL-3 cntainment labs shuld have: Class II Bilgical Safety Cabinets Each unit shuld have an alarm at the discharge f the hd that will sund if the CFM drps belw 250. A Class II BSC is designed with inward air flw at a velcity t prtect persnnel ( linear feet per minute); high-efficiency particulate air (HEPA) filtratin f the air flw dwn acrss the wrk surface (vertical laminar flw) fr prtectin f the research material; and HEPA-filtered exhaust air fr envirnmental prtectin. Page 118 f 201

119 Persnnel Training a. Waste cntainer inside BSCs If a small plastic pail is used, it cntains a bihazard bag as a liner. Radiactive bihazardus waste will be cllected in a separately labeled cntainer when radiactivity is used. b. Aspiratr flasks in BSCs All liquid wastes (e.g., cell culture supernatants) are cllected inside the BSC an aspiratr flask cntaining the apprpriate decntaminant (e.g., bleach, 10% final cncentratin) and labeled accrdingly. The cntainers can be discnnected frm the aspiratin unit by quick snap, dublesealing cnnectins (see Waste Sectin). The vacuum line is prtected by a HEPA filter. Waste cntainers utside the BSCs A hands free sink a. All articles, such as paper and plastic wrappers f serlgical pipettes, are cllected in a cvered waste cntainer lined with duble bihazard bags n the flr adjacent t the BSCs. An additinal sink, preferably with an autmatic r ft perated faucet, shuld be placed in the anterm. A means fr the decntaminatin f waste materials prir t remval frm the facility Phne, fax, and cmputer capability Anterm with clean supplies, such as lab gwns, masks, glves, plastic bags, sap, and paper twels, alng with the visitr lgbk. Fr pest cntrl, a pest management plan must in place prir t peratins. All bisafety cabinets, incubatrs, and -70 C freezers are supplied with emergency pwer. The training fr BSL-3 facilities must address varius cnstituencies, including: Labratry wrkers with cntact with the bilgical materials Animal facility wrkers Page 119 f 201

120 General emplyees (i.e., nn-labratry wrkers) Facilities and maintenance wrkers Institutinal first respnders First respnders frm external agencies The purpse f a cmprehensive training prgram is t ensure emplyees are aware f the hazards t which they might be expsed and are familiar with safety wrk prcedures and hw t prtect themselves, their clleagues, and the envirnment frm ptential expsure. Site-specific training fr BSL-3 facilities is an IBC mandate and requires additinal training n the facility SOPs. The labratry directr is respnsible fr this training and shuld cntact the EHS fr assistance if needed. The tpics t be included in the training prgram will vary depending n the bilgical materials used and the types f research cnducted. At a minimum, hwever, the fllwing areas shuld be addressed: Bilgy f rganism used In the case f ABSL-3 wrk persnnel must have apprpriate animal handling training Use within facility and standard precautins Disinfectants Recrd keeping and data entry Lab inspectins and husekeeping duties Emergency respnse Fire cntrl and suppressin Medical emergencies Entry/exit requirements and ther administrative measures Safety equipment; primary and secndary barriers Receipt and inspectin f cultures Transprt f agents within the labratry Strage, segregatin, and recrd-keeping in labratry Remval f materials frm facility Page 120 f 201

121 Spill cntrl Utility discnnectin Cntrl panels, functin and shut ff Equipment malfunctins Reprting Inspectin and Maintenance f Cntainment Facility The purpse f labratry inspectin is t ensure that the wrking envirnment is safe, all pieces f equipment are functining prperly, and that administrative cntrls are bserved. A rbust pstapprval mnitring prgram is designed t identify prblems early and t develp preventive measures. The scpe and frequency f the inspectins will depend n the specific agents used, type f activities cnducted, and the labratry recrd f maintaining a safe and cmpliant facility. In general, it will fllw the Quarterly Audit Prcedures (see Appendix V, IBC Oversight Prgram) and will have additinal site-specific elements added. Prcedures fr Perfrming Preventative Maintenance The prper perfrmance and maintenance f equipment in any labratry is imprtant, and in a BSL-3 facility, it is f paramunt imprtance. Engineering cntrls are a significant cmpnent f the facility s verall safety design. Therefre, a cmprehensive preventive maintenance and inspectin prgram is critical t the safe peratins f such facilities. The specific scpe and extent f the preventive maintenance and equipment inspectin will be dependent n the facility and its design. Hwever, the fllwing is a list f minimum recmmendatins: Equipment Maintenance The fllwing physical plant equipment is checked and inspected n a rutine basis: exhaust fans and ther HVAC equipment autclave vacuum lines emergency generatr Nte: Specific schedules are develped fr each facility. Page 121 f 201

122 Checking these devices cnsists f bserving whether the device is perating nrmally. Inspecting these devices cnsists f visually inspecting the device s integral parts, such as seals, bearings, wiring, pwer supply and brushes, and replacing/repairing defective parts. Equipment lcated inside the cntainment lab will be inspected in cnjunctin with the annual recertificatin. Any piece f equipment needing repair at any ther time will be decntaminated befre a mechanic is allwed entry. The area arund the equipment will als be decntaminated and all infectius materials will be secured. The mechanics will fllw facility entry and exit prcedures and be escrted by facility persnnel. All tls used as part f repair and maintenance will be decntaminated befre they are remved frm the labratry. Bilgical Safety Cabinet Maintenance Each unit is inspected annually by a qualified individual t dcument perfrmance and efficiency and t prvide annual certificatin. The integrity f the HEPA filter is als tested. Emergency Equipment Testing Inspect the emergency equipment fllwing the instructins belw: Emergency Generatr Alarms a. Discnnect the labratry/generatr cntact. b. Next, lcate the emergency test run switch and turn t test psitin. c. The generatr mtr shuld begin t turn ver. d. When the engine starts, run the generatr fr 15 minutes and then turn the test switch t ff. e. Reset the labratry/generatr cnnectin t n and return the test run switch t aut. a. Check the pressure-sensitive alarm at least biannually. b. Fire alarm is checked mnthly and inspected annually. Emergency Shwers and Eye Wash Statins Page 122 f 201

123 a. Check the eye wash statins weekly by turning n the flw f water and watching fr adequate delivery that is clean and free f debris r rust. b. Shwers and eyewash statins are inspected every six mnths fr prper peratin and flw rate. A lg is maintained. Fire Extinguishers Wrking with Animals at ABSL-3 a. Inspected mnthly b. Decntaminated and replaced annually. ABSL-3 prtcls are reviewed by the BU IACUC and IBC cmmittees t assure bth animal welfare and wrker safety and health cncerns. Standard practice fr wrking safely with animals at BU has been established in Appendix N f this manual. Specific practices fr wrking with animals at ABSL-2 are currently utlined in Appendix O f this manual. Wrking with animals at BSL-3 builds n the practices utlined in the abve mentined appendices. Additinal practices at ABSL-3 include the fllwing: Standard Practices Specific requirements established in the IACUC and IBC apprved prtcls must be dcumented and fllwed. All persnnel are trained n the specific hazards f the prject and are required t read and fllw facility specific prcedures. Training is prvided t animal care and labratry persnnel cncerning animal husbandry prcedures, ptential hazards, and manipulatins f infectius agents and the hazards assciated with them. All labratry and animal care persnnel must participate in the ROHP medical surveillance prgram utlined in Appendix R. Standard Micrbilgical Practices Apprpriate signage must be placed utside the areas where infectius animals and/r materials are hused and manipulated. All persnnel handling animals at BSL-3 must wear facility specific PPE utlined in facility specific SOP s. All prcedures must be perfrmed in a way s as t minimize the creatin f aersls r splatters f infectius materials. Mechanical pipetting devices must be used. Page 123 f 201

124 Sharps must be handled in accrdance with Chapter 9 f this manual. Dispsable needles must nt be bent, brken, sheared, recapped r remved frm dispsable syringes. As described in Chapter 9 all dispsable needles must be placed in sharps cntainers fr dispsal. Sharps cntainers shuld be placed as clse t the wrk area as pssible. Brken glassware shuld never be handled directly. Rather it shuld be cleaned up using a brush and dustpan, and/r tngs r frceps. Plastic wear shuld be substituted where pssible. All wastes frm the animal rm (including animal tissues, carcasses, and bedding) must be transprted frm the animal rm in leak-prf cntainers fr decntaminatin and apprpriate dispsal. Special Practices See special practices in the BSL-3 sectin When a prcedure cannt be perfrmed in a BSC, a cmbinatin f PPE and/r ther cntainment devices must be used. Such prcedures are reflected in the Facility specific SOP s. Restraint devices and practices will be used t reduce the risk f expsure during animal manipulatins. Animals will be hused in cntainment caging systems that are specific t a facility in rder t reduce the risk f infectius aersls frm infected animals r their bedding. Ventilated caging systems must be designed t prevent cntaminatin utside f the cage. Safety mechanisms must be in place t prevent exhaust plenums frm becming psitive, and the system shuld be alarmed t indicate malfunctins. All infectius materials are decntaminated prir t exiting the facility. Once decntaminated, all cages and materials are handled by Bstn University Animal Science Center (BUASC). Decntaminated Cages are als washed in a mechanical cage washer with a final rinse temperature f 180 degrees Farenheit. Safety Equipment (Primary Barriers and PPE) See safety equipment in the BSL-3 sectin Persnnel handling animals in the BSL-3 facility must gwn accrding t facility prcedures. Dispsable PPE must be remved when exiting the area where infected animals and materials are hused. Page 124 f 201

125 All dispsable materials are decntaminated by autclave prir t leaving the facility. Glves must be wrn and are indicated in facility specific SOP s. Labratry Facilities Glves are t be changed when cntaminated r therwise cmprmised. Glves must be changed befre exiting animal area and must be remved in a manner s as t prevent the spread f infectius materials. Dispsable glves are nt t be washed r reused. See Labratry Facilities in the BSL-3 sectin Drs where infectius animals r materials are hused will pen inward and are self-clsing and are never prpped pen. The interir surfaces f the animal areas must be water resistant s as t be able t be cleaned and decntaminated. Directin f airflw in the animal facility is inward and shuld be in accrdance with the Guide fr Care and Use f Labratry Animals. Page 125 f 201

126 Frm B1 Certificatin and Maintenance f BSL-3 Labratries MAINTENANCE BSL-3 LAB QUARTERLY MAINTENANCE Subject: CHECKLIST Date: Time: Temp: Humidity: ITEM SUBJECT Pass Fail Cmment/Crrectin 1 Exhaust Fan N. a Belts b Pulleys c Hardware d Damper & Actuatr e Lubricate Drive Bearings f Electrical Cnnectins g Mtr Vltage h Mtr Amperage 2 Exhaust Fan N. a Belts b Pulleys c Hardware d Damper & Actuatr e Lubricate Drive Bearings f Electrical Cnnectins g Mtr Vltage h Mtr Amperage 3 VFD N. a Electrical Cnnectins b Frequency c % d Vltage e Amperage Page 126 f 201

127 MAINTENANCE ITEM SUBJECT Pass Fail Cmment/Crrectin 4 VFD N. a Electrical Cnnectins b Frequency c % d Vltage e Amperage 5 HEPA Filter N. a P b Change Filter PI/B&V 6 HEPA Filter N. a P b Change Filter PI/B&V 7 Bisafety Cabinets a Recertificatin PI/B&V 8 Bisafety Cabinets a Recertificatin PI/B&V All unacceptable items must be described and crrective actin listed. Page 127 f 201

128 Appendix G Bisafety Level 4 (BSL-4) Requirements Use f Bisafety Level 4 (BSL-4) agents is subject t very strict federal, state, and lcal requirements and can nly be undertaken if the Institutinal Bisafety Cmmittee has reviewed the applicatin, facility design, and specific wrk practices and apprved the prject. The fllwing guidelines are intended t prvide the investigatrs a basis fr such applicatin preparatin and are nt intended t cnstitute a cmprehensive, prject-specific guide. Each facility will have a site-specific set f perating prcedures that must be apprved by the IBC. A BSL-4 facility is required fr wrk with high hazard and extic agents that pse a high individual risk f aersl-transmitted labratry infectins and life-threatening disease. Nte: BSL-4 facilities must develp a site-specific bisafety manual based n the general utlines in this appendix. General requirements Individuals wrking in such facilities must have specific and thrugh training in: Handling f extremely high hazard infectius agents Primary and secndary cntainment requirements Standard and special practices The cntainment equipment and the labratry design characteristics In additin, they are supervised by cmpetent scientists wh are trained and experienced in wrking with these agents. Access t the labratry is strictly cntrlled by the labratry directr. Access t the facility is separate frm the general building access. A specific facility peratins manual is prepared r adpted. Within wrk areas f the facility: All activities are cnfined t Class III bilgical safety cabinets, r Class II bilgical safety cabinets used with ne-piece psitive pressure persnnel suits ventilated by a life-supprt system. The BSL-4 labratry has special engineering and design features t prevent micrrganisms frm being disseminated int the envirnment. Page 128 f 201

129 Labratry persnnel are required t be medically cleared by the Research Occupatinal Health Prgram prir t wrking in a BSL 3 labratry (See Appendix R Medical Surveillance Prgram) where apprpriate testing and immunizatins will be perfrmed based n ccupatinal expsure and agents handled. Training The purpse f a cmprehensive training prgram is t ensure that the emplyees are aware f the hazards t which they might be expsed and are familiar with safety wrk prcedures and hw t prtect themselves, their clleagues, and the envirnment frm ptential expsure. The training requirements fr BSL-4 facilities will include all f the categries listed in BSL-3. Additinal site-specific training fr BSL-4 facilities is an IBC mandate and requires additinal training in the facility standard perating prcedures (SOP). Training fr BSL-4 facilities must address varius cnstituencies: Labratry wrkers with cntact with the bilgical materials Animal facility wrkers General emplyees (i.e., nn-labratry wrkers) Facilities and maintenance wrkers Institutinal first respnders First respnders frm external agencies EHS will crdinate all training related t safety. The labratry directr is respnsible fr specific lab related training and shuld cntact the BSO fr assistance if needed. The facility will als have rutine emergency and evacuatins drills and site-specific SOP fr safety and security. Standard Micrbilgical Practices Access t the labratry is limited by Public Safety, EHS, RO, the Principal Investigatr, Labratry Directr r designee when experiments are in prgress. Plicies fr safe handling f sharps are instituted. All prcedures are perfrmed carefully t minimize the creatin f aersls. Wrk surfaces are decntaminated at least nce a day and after any spill f viable material. Page 129 f 201

130 All waste is decntaminated befre dispsal by an apprved methd, such as autclaving. A pest cntrl prgram is in effect. Only persns whse presence in the facility r individual labratry rms is required fr prgram r supprt purpses are authrized t enter. The supervisr has the final respnsibility fr assessing each circumstance and determining wh may enter r wrk in the labratry. Access t the facility is limited by means f secure, lcked drs; accessibility is managed by the labratry directr, BSO, r ther persn respnsible fr the facility s physical security. Befre entering, persns are advised f the ptential bihazards and instructed as t apprpriate safeguards fr ensuring their safety. Authrized persns cmply with the instructins and all ther applicable entry and exit prcedures. An entry/exit cntrl lg is in effect either via an electrnic access cntrl fr each individual r a lgbk, signed by all persnnel, indicating the date and time f each entry and exit. Practical and effective prtcls fr emergency situatins are established. When infectius materials r infected animals are present in the labratry r animal rms, hazard warning signs, incrprating the universal bihazard symbl, are psted n all access drs. The sign identifies the agent, lists the name f the labratry directr r ther respnsible persn(s), and indicates any special requirements fr entering the area (e.g., the need fr immunizatins r respiratrs). Labratry persnnel are ffered r receive available immunizatins as necessary fr the agents handed r ptentially present in the labratry. A bisafety manual is prepared r adpted. Persnnel are advised f special hazards and are required t read and fllw instructins n practices and prcedures. Labratry and supprt persnnel receive apprpriate training n the ptential hazards assciated with the wrk invlved, the necessary precautins t prevent expsures, and the expsure evaluatin prcedures. Persnnel receive annual updates r additinal training as necessary fr prcedural changes. Persnnel enter and leave the labratry nly thrugh the clthing change and shwer rms. They take a decntaminating shwer each time they leave the labratry. Persnnel use the airlcks t enter r leave the labratry nly in an emergency. Page 130 f 201

131 Persnal clthing is remved in the uter clthing change rm and kept there. Supplies and materials needed in the facility are brught in by way f the duble-dr autclave, fumigatin chamber, r airlck, which is apprpriately decntaminated between each use. After securing the uter drs, persnnel within the facility retrieve the materials by pening the interir drs f the autclave, fumigatin chamber, r airlck. These drs are secured after materials are brught int the facility. A high degree f precautin must always be taken with any cntaminated sharp items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels. Needles and syringes r ther sharp instruments are restricted in the labratry fr use nly when there is n alternative, such as fr parenteral injectin, phlebtmy, r aspiratin f fluids frm labratry animals and diaphragm bttles. Plasticware shuld be substituted fr glassware whenever pssible. Only needle-lcking syringes r dispsable syringe-needle units (i.e., needle is integral t the syringe) are used fr injectin r aspiratin f infectius materials. Used dispsable needles must nt be bent, sheared, brken, recapped, remved frm dispsable syringes, r therwise manipulated by hand befre dispsal; rather, they must be carefully placed in cnveniently lcated puncture-resistant cntainers used fr sharps dispsal. Nn-dispsable sharps must be placed in a hard-walled cntainer fr transprt t a prcessing area fr decntaminatin, preferably by autclaving. Syringes that re-sheathe the needle, needleless systems, and ther safety devices are used when apprpriate. Brken glassware must nt be handled directly by hand, but must be remved by mechanical means, such as a brush and dustpan, tngs, r frceps. Cntainers f cntaminated needles, sharp equipment, and brken glass must be decntaminated befre dispsal, accrding t any lcal, state, r federal regulatins. Bilgical materials in a viable r intact state must be remved in accrdance with a prtcl that has been apprved by the IBC. Bilgical materials remved frm the labratry are transferred t a nn-breakable, sealed cntainer. All materials (except bilgical materials that are t remain in a viable r intact state) must be remved frm the BSL-4 labratry nly after they have been autclaved r decntaminated. Page 131 f 201

132 Equipment r material that might be damaged by high temperatures r steam may be decntaminated by gaseus r vapr methds in an airlck r chamber designed fr this purpse. Labratry equipment is decntaminated rutinely after wrk with infectius materials is finished, and especially after vert spills, splashes, r ther cntaminatin with infectius materials. Equipment is decntaminated befre it is sent fr repair r maintenance. Apprpriate prfessinal staff r thers prperly trained and equipped t wrk with cncentrated infectius material cntain and clean up spills f infectius materials. A spill prcedure is develped and psted within the labratry. A system must be established fr reprting labratry accidents and expsures and emplyee absenteeism, and fr the medical surveillance f ptential labratry-assciated illnesses. Written recrds must be maintained. An essential adjunct t such a reprting-surveillance system is the availability f a facility fr the quarantine, islatin, and medical care f persnnel with ptential r knwn labratry-assciated illnesses. Materials nt related t the experiment being cnducted (e.g., plants, animals, and clthing) are nt permitted in the facility. Primary Barriers All prcedures within the facility are cnducted in the Class III bilgical safety cabinet r in Class II bilgical safety cabinets used in cnjunctin with ne-piece psitive pressure persnnel suits ventilated by a life-supprt system. Secndary Barriers There are tw mdels fr BSL-4 labratries: The cabinet labratry, where all handling f the agent is perfrmed in a Class III bilgical safety cabinet, and The suit labratry, where persnnel wear a prtective suit. Cabinet Labratry BSL-4 labratries may be based n either mdel r a cmbinatin f bth mdels in the same facility. If a cmbinatin is used, each type must meet all the requirements identified fr that type. Daily inspectins f all cntainment parameters (e.g., directinal airflw) and life supprt systems are cmpleted befre labratry wrk is initiated t ensure the labratry is perating accrding t its perating parameters. Page 132 f 201

133 Walls, flrs, and ceilings f the cabinet rm and inner change rm are cnstructed t frm a sealed internal shell that facilitates fumigatin and is resistant t entry and exit f animals and insects. Flrs are integrally sealed and cved. The shell s internal surfaces are resistant t liquids and chemicals t facilitate the area s cleaning and decntaminatin. All penetratins in these structures and surfaces are sealed. Openings arund drs int the cabinet rm and inner change rm are minimized and are capable f being sealed t facilitate decntaminatin. Any windws are breakage-resistant and sealed. Any drains in the cabinet rm flr are cnnected directly t the liquid waste decntaminatin system. Sewer vents and ther service lines cntain filters and prtectin against vermin. Bench tps have seamless r sealed surfaces that are impervius t water and are resistant t mderate heat and the rganic slvents, acids, alkalis, and chemicals used t decntaminate wrk surfaces and equipment. Labratry furniture is f simple, pen cnstructin, capable f supprting anticipated lading and uses. Spaces between benches, cabinets, and equipment are accessible fr cleaning and decntaminatin. Chairs and ther furniture used in labratry wrk shuld be cvered with a nn-fabric material that can be easily decntaminated. A hands-free r autmatically perated handwashing sink is prvided near the dr f the cabinet rm(s) and the uter and inner change rms. Duble-dr autclaves are prvided fr decntaminating materials passing ut f bth the Class III bilgical safety cabinet(s) and the cabinet rm(s). Autclaves that pen utside f the cntainment barrier must be sealed t the wall f the cntainment barrier. The autclave drs are autmatically cntrlled s that the utside dr can nly be pened after the autclave sterilizatin cycle has been cmpleted. Pass-thrugh dunk tanks, fumigatin chambers, r equivalent decntaminatin methds are prvided s that materials and equipment that cannt be decntaminated in the autclave can be safely remved frm bth the Class III bilgical safety cabinet(s) and the cabinet rm(s). The facility must be tested fr verificatin that the design and peratinal parameters have been met prir t peratin. Facilities shuld be re-verified annually against these. Page 133 f 201

134 Suit Labratry The BSL-4 facility cnsists f either a separate building r a clearly demarcated and islated zne within a building. The rms in the facility are arranged t ensure passage thrugh the changing and decntaminatin areas prir t entering the rm(s) where wrk is dne with BSL-4 agents (suit area). Outer and inner change rms, separated by a shwer, are prvided fr persnnel entering and leaving the suit area. A specially designed suit area is maintained in the facility t prvide persnnel prtectin equivalent t that prvided by Class III bilgical safety cabinets. Persnnel wh enter this area wear a ne-piece psitive pressure suit that is ventilated by a life-supprt system prtected by HEPA filtratin. The life-supprt system includes redundant breathing air cmpressrs, alarms, and emergency backup breathing air tanks. Entry t this area is thrugh an airlck fitted with airtight drs. A chemical shwer is prvided t decntaminate the surface f the suit befre the wrker leaves the area. An autmatically starting emergency pwer surce is prvided, at a minimum, fr the exhaust system, life-supprt systems, alarms, lighting, entry and exit cntrls, and bilgical safety cabinets. The air pressure within the suit is psitive t the surrunding labratry. The air pressure within the suit area is lwer than that f any adjacent area. Emergency lighting and cmmunicatin systems are prvided. All penetratins int the internal shell f the suit area, chemical shwer, and airlcks are sealed. A daily inspectin f all cntainment parameters (e.g., directinal airflw, chemical shwers) and lifesupprt systems is cmpleted befre labratry wrk is initiated t ensure that the labratry is perating accrding t its perating parameters. A duble-dr autclave is prvided at the cntainment barrier fr decntaminating waste materials t be remved frm the suit area. The autclave dr, which pens t the area external t the suit area, is sealed t the uter wall f the suit area and is autmatically cntrlled s that the utside dr can be pened nly after the autclave sterilizatin cycle. A dunk tank, fumigatin chamber, r ventilated airlck fr decntaminatin is prvided fr passage f materials, supplies, r equipment that is nt brught int the suit area thrugh the change rm. These devices can be als used fr the safe remval f materials, supplies, r equipment frm the labratry that cannt be decntaminated in the autclave. The suit area s walls, flrs, and ceilings are cnstructed t frm a sealed internal shell. Page 134 f 201

135 The shell s internal surfaces are resistant t liquids and chemicals, facilitating the area s cleaning and decntaminatin. All penetratins in these structures and surfaces are sealed. Any drains in the flr f the suit area cntain traps filled with a chemical disinfectant f demnstrated efficacy against the target agent, and they are cnnected directly t the liquid waste decntaminatin system. Sewer vents and ther service lines cntain filters. Internal facility appurtenances in the suit area, such as light fixtures, air ducts, and utility pipes, are arranged t minimize the hrizntal surface area. Bench tps have seamless surfaces that are impervius t water and resistant t mderate heat and the rganic slvents, acids, alkalis, and chemicals used t decntaminate the wrk surfaces and equipment. Labratry furniture is f simple, pen cnstructin capable f supprting anticipated lading and uses. Nn-prus materials are preferable. Spaces between benches, cabinets, and equipment are accessible fr cleaning and decntaminatin. Chairs and ther furniture used in labratry wrk shuld be cvered with a nn-fabric material that can be easily decntaminated. A hands-free r autmatically perated handwashing sink is prvided in the suit area(s); handwashing sinks in the uter and inner change rms shuld be cnsidered based n the risk assessment. Any windws are breakage-resistant and are sealed. Liquid effluents frm sinks, flr drains (if used), autclave chambers, and ther surces within the cntainment barrier are decntaminated by a prven methd, preferably heat treatment, befre being discharged t the sanitary sewer. A dedicated, nn-recirculating ventilatin system is prvided. The system s supply and exhaust cmpnents are balanced t ensure directinal airflw frm the area f least hazard t the area(s) f greatest ptential hazard. Redundant supply fans are recmmended, and redundant exhaust fans are required. The differential pressure/directinal airflw between adjacent areas is mnitred and alarmed t indicate a system malfunctin. An apprpriate visual pressure mnitring device that indicates and cnfirms the pressure differential f the suit area must be prvided and lcated at the entry t the clean change rm. The airflw in the supply and exhaust cmpnents is mnitred, and an HVAC cntrl system is installed t prevent psitive pressurizatin f the labratry. The supply air t the suit area, decntaminatin shwer, and decntaminatin airlck is prtected by passage thrugh a HEPA filter. The general rm exhausts air frm the suit area, decntaminatin Page 135 f 201

136 shwer, and decntaminatin airlck, and the air is treated by a passage thrugh tw HEPA filters in series prir t discharge t the utside. The air is discharged away frm ccupied spaces and air intakes. The HEPA filters are lcated as near as practicable t the surce in rder t minimize the length f ptentially cntaminated ductwrk. All HEPA filters need t be tested and certified annually. The HEPA filter husings are designed t allw fr in situ decntaminatin f the filter prir t remval. Alternatively, the filter can be remved in a sealed, gas-tight primary cntainer fr subsequent decntaminatin and/r destructin by incineratin. Design f the HEPA filter husing shuld facilitate validatin f the filter installatin. The use f pre-certified HEPA filters can be an advantage. The service life f the exhaust HEPA filters can be extended thrugh adequate pre-filtratin f the supply air. The psitining f the supply and exhaust pints shuld be such that dead air space in the suit rm is minimized. The treated exhaust air frm Class II bilgical safety cabinets, lcated in a facility where wrkers wear psitive pressure suits, may be discharged int the rm envirnment r t the utside thrugh the facility air exhaust system. If the treated exhaust is discharged t the utside thrugh the facility exhaust system, it is cnnected t this system in a manner that avids any interference with the air balance f the cabinets r the facility exhaust system. The BSL-4 facility design and peratinal prcedures must be dcumented. The facility must be tested fr verificatin that the design and peratinal parameters have been met prir t peratin. Facilities shuld be re-verified annually against these prcedures as mdified by peratinal experience. Apprpriate cmmunicatin systems shuld be prvided between the labratry and the utside. Page 136 f 201

137 Appendix H Guidelines fr Wrk with Txins f Bilgical Origin This categry f txins is defined as any txic substance f natural rigin prduced by an animal, plant, r micrbe. They are nn-vlatile, usually nt dermally active (myctxins are an exceptin), and tend t be mre txic per weight than many chemical agents. Wrking with bilgically-derived txins may present health risks due t rutes f expsure that are nt always taken int cnsideratin. The labratry facilities, equipment, and prcedures fr wrk with txins must reflect the intrinsic level f hazard psed by a particular txin, as well as the ptential risks inherent in the peratins being perfrmed. The IBC will evaluate all such uses and determine if the prpsed safeguards are adequate r nt. Nte: Txins listed in the select agents rule must be belw the excluded amunts if they are t remain exempt frm the regulatins. Otherwise they are subject t the full select agent rules. The fllwing infrmatin is taken frm the CDC/NIH s Bisafety in Micrbilgical and Bimedical Labratries (BMBL) (5 th Editin, Bisafety guidelines are intended t prvide general safety requirements fr the use f such txins. General Infrmatin The labratry facilities, equipment, and prcedures apprpriate fr wrk with txins f bilgical rigin must reflect the intrinsic level f hazard psed by a particular txin, as well as the ptential risks inherent in the peratins perfrmed. If bth txins and infectius agents are used, bth must be cnsidered when cntainment equipment is selected and plicies and prcedures are written. If animals are used, animal safety practices must als be cnsidered. Standard Practices Prvide all labratry persnnel with training specific fr the txins being used. Maintain an accurate inventry. Stre txin stcks in lcked strage rms, cabinets, r freezers. Prepare primary cntainers f txin slutins and handle primary cntainers f dry frms f txins in a chemical fume hd, a glve bx, r a bilgical safety cabinet r equivalent cntainment system. Whenever pssible, recnstitute entire vial f pwdered txin by injecting diluent thrugh septum. Glve selectin If pwdered txin must be handled, select glves that d nt generate static electricity. D nt use latex. Page 137 f 201

138 When handling txins that are percutaneus hazards (irritants, necrtic t tissue, r extremely txic frm dermal expsure), select glves that are knwn t be impervius t the txin. In additin, standard practices listed under BSL-2 and/r BSL-3 shuld be reviewed and incrprated as apprpriate int prtcls fr wrk with txins. Special Practices Specific special practices will depend n a number f parameters, such as the txin used r the type f peratins perfrmed. In general: Each labratry shuld develp a chemical hygiene plan specific t the txin(s) used in that labratry. The chemical hygiene plan shuld: Identify the hazards that will be encuntered in nrmal use f the txin and thse that culd be encuntered in case f a spill r ther accident, and Specify the plicies and practices t be used t minimize risks (e.g., cntainment and persnal prtective equipment, management f spills, management f accidental expsures, medical surveillance). Training specific t the txin(s) used shuld be required and dcumented fr all labratry persnnel wrking with txins, befre starting wrk with the txin, and at intervals thereafter. An inventry cntrl system shuld be in place. Txins shuld be stred in lcked strage rms, cabinets, r freezers when nt in use. Access t areas cntaining txins shuld be restricted t thse whse wrk assignments require access. Preparatin f primary cntainers f txin stck slutins and manipulatins f primary cntainers f dry frms f txins shuld be cnducted in a chemical fume hd, a glve bx, r a bilgical safety cabinet r equivalent cntainment system apprved by the safety fficer. HEPA and/r charcal filtratin f the exhaust air may be required, depending n the txin. The user shuld verify inward airflw f the hd r bilgical safety cabinet befre initiating wrk. All wrk shuld be dne within the peratinally effective zne f the hd r bilgical safety cabinet. When txins are in use, the rm shuld be psted t indicate Txins in Use - Authrized Persnnel Only. Any special entry requirements shuld be psted n the rm s entrance(s). Page 138 f 201

139 Safety Equipment Only persnnel whse presence is required shuld be permitted in the rm while txins are in use. All high-risk peratins shuld be cnducted with tw knwledgeable individuals present. Each must be familiar with the applicable prcedures, maintain visual cntact with the ther, and be ready t assist in the event f an accident. Befre cntainers are remved frm the hd, cabinet, r glve bx, the exterir f the clsed primary cntainer shuld be decntaminated and placed in a clean secndary cntainer. Txins shuld be transprted nly in leak-/spill-prf secndary cntainers. Cntaminated and ptentially cntaminated prtective clthing and equipment shuld be decntaminated using methds knwn t be effective against the txin befre remval frm the labratry fr dispsal, cleaning, r repair. If decntaminatin is nt pssible/practical, materials (e.g., used glves) shuld be dispsed f as txic waste. Materials cntaminated with infectius agents, as well as txins, shuld als be autclaved r therwise rendered nninfectius befre leaving the labratry. The interir f the hd, glve bx, r cabinet shuld be decntaminated peridically, fr example, at the end f a series f related experiments. Until decntaminated, the hd, bx, r cabinet shuld be psted t indicate that txins are in use, and access t the equipment and apparatus restricted t necessary, authrized persnnel. The IBC will review each prtcl and determine the specific safety requirements. In general, txins are expected t meet all BSL-2 requirements and, in sme instances, might require BSL-3 level safety equipment. When using an pen-frnted fume hd r bilgical safety cabinet, prtective clthing, including glves and a dispsable, lng-sleeved bdy cvering (gwn, labratry cat, smck, cverall, r similar garment) shuld be wrn s that hands and arms are cmpletely cvered. Eye prtectin shuld be wrn if an pen-frnted cntainment system is used. Other prtective equipment may be required, depending n the characteristics f the txin and the cntainment system. Fr example, use additinal respiratry prtectin if aersls may be generated and it is nt pssible t use cntainment equipment r ther engineering cntrls. When handling dry frms f txins that are electrstatic: D nt wear glves (such as latex) that help t generate static electricity. Use a glve bag within a hd r bilgical safety cabinet, a glve bx, r a Class III bilgical safety cabinet. When handling txins that are percutaneus hazards (irritants, necrtic t tissue, r extremely txic frm Page 139 f 201

140 dermal expsure), select glves that are knwn t be impervius t the txin. Cnsider bth txin and diluents when selecting glves and ther prtective clthing. If infectius agents and txins are used tgether in an experimental system, cnsider bth when selecting prtective clthing and equipment. Labratry Facilities Labratry facility recmmendatins listed under BSL-2 and BSL-3, as well as OSHA standards, shuld be reviewed and incrprated as apprpriate int prtcls fr wrk with txins. When vacuum lines are used with systems cntaining txins, they shuld be prtected with a HEPA filter t prevent entry f txins int the lines. Sink drains shuld be similarly prtected when water aspiratrs are used. Receiving, Inspecting, Strage and Distributin f Samples/Materials The labratry must develp specific standard perating prcedures (SOP) that addresses safety receipt, inspectin, strage, usage and dispsal f the txin used. IBC requires that all SOPs develped incrprate the safety and persnal prtective equipment int the prcedures. Page 140 f 201

141 Appendix I List f Bilgical Agents with the Ptential t Cause Labratry Acquired Infectin (LAI) in use at Bstn University The List f Bilgical Agents with the Ptential t Cause Labratry Acquired Infectin (LAI) cntains sme BSL-3 agents and many BSL-2 agents in use within the Bstn University/Bstn Medical Center research cmmunity. Principal Investigatrs and research staff listed n apprved IBC prtcls invlving these bilgical agents with the ptential t cause LAI must receive agent specific training; agent specific identificatin cards t be carried by thse persnnel; and Agent Infrmatin Sheets (AIS) prviding safety and handling instructins. The list f bilgical agents with the ptential t cause LAI is dynamic and will be rutinely reviewed fr pathgens that researchers are prpsing t use. As new agents apprved by the IBC are intrduced int the labratry envirnment, they may be added t the list r as agents are n lnger being used they will be remved frm the list. Cntact the IBC Office at ibc@bu.edu fr the current list. Page 141 f 201

142 Appendix J Prin Research/Creutzfeldt-Jacb Disease (CJD) Guidelines Creutzfeldt-Jacb Disease (CJD) is ne f a grup f diseases called transmissible spngifrm encephalpathies that als includes kuru and Gerstmann-Sträussler-Scheinker syndrme f humans, scrapie f sheep, and mad cw disease f cattle and dairy cws. Lng (mnths t years) incubatin perids precede the nset f clinical illness, with chrnic prgressive pathlgy that may last weeks t mnths. CJD is characterized by prgressive dementia, myclnic fasciculatins, ataxia, and smnlence. The clinical curse f CJD usually lasts several mnths and is invariably fatal. N effective treatment is available, and there are n knwn cases f remissins r recveries. Transmissible spngifrm encephalpathies are caused by a unique infectius agent called a prin, cmpsed f a prteinaceus material devid f detectable amunts f nucleic acid. Prins are unusually resistant t standard means f inactivatin, including frmaldehyde, alchl, and UV radiatin. Hwever, they can be inactivated by fresh husehld bleach; 1.0N sdium hydrxide (NaOH); 4.0N guanidine reagents; phenl; and autclaving. Prcedures invlving brain tissue frm patients with neurlgical degenerative disrders (such as CJD and Alzheimer s disease) pse special challenges in reducing ptential expsure t prins; such material shuld be handled with at least the same precautins as HIVpsitive r HBV-psitive human tissue. The CDC classifies prins as Risk Grup 2 agents requiring Bisafety Level 2 (BSL-2) cntainment. BU/BMC require all researchers wrking with these agents t have a valid Bilgical Use Apprval; IBC apprval must be granted befre any research can be initiated. The fllwing current BU/BMC prin prcedures have evlved frm best practices used at many institutins, including the University f Califrnia, San Francisc and the University f Califrnia, San Dieg, that have been engaged in such research fr years. Nte: Wrk with bvine spngifrm encephalpathy falls under the select agents rule. Safety Prcedures Prin agent must be treated as bihazardus. An autclave fr treatment f slid wastes is present. All equipment that have cme in cntact with the agent, packaging, cntainers and all unused prtins and derivatives frm the agent will be treated befre dispsal. All fixed, nn-fixed, r frzen tissues that cntain the agent must be placed within watertight cntainers and labeled with the universal bihazard symbl and the ntatin Infectius Materials. Treatment can be dne with sdium hypchlrite at 20,000 ppm available chlrine fr 1 hur at 20 0 C. 20, 000 ppm is prepared using 40% husehld bleach (which is 5.25% sdium hypchlrite) and 60% diluents. Treatment may als be dne with 1N NaOH slutin fr 1 hur at 20 0 C. 1N r 4% slutin is prepared using 24 z NaOH in 1 galln f water. Page 142 f 201

143 Prus r slid waste autclaving is dne fr C t C at 30 psi fr 18 minutes (hlding time and temperature) r 6 separate cycles at C t C at 30 psi fr 3 minutes (hlding time and temperature). Persnnel wrking with the agent must nt have cntact with any animal clnies withut IACUC apprval in the labratry cmplex r with susceptible animal species. Persnnel must wear glves and gwns while handling tissues that are ptentially cntaminated. All prtective clthing must be remved befre leaving the labratry. Persnnel wrking with the material are instructed n the prcedures fr handling the agent. Snicatin r hmgenizatin f tissues must be perfrmed in a prperly certified Class II Bilgical Safety Cabinet (BSC). Micrtme blades and knives used fr cutting r sectining tissue sample must be cleaned with an instrument that des nt put the peratr s hand r finger in r near cntact with the blade. Any skin cntact with pssibly infectius materials shuld be fllwed by washing with 1N sdium hydrxide fr tw t three minutes, fllwed by extensive washing with water. The PI must cntact the Bisafety Officer in writing regarding spills and accidents that result in vert expsure t tissues. The reprt must include the fllwing: Recrds Specificatin f amunt released, time invlved, and explanatin f prcedures used t determine the amunt invlved. Descriptin f the area invlved and the extent f emplyee expsure. Reprt f medical treatment prvided. Crrective actin taken t prevent the reccurrence f the incident. ROHP must maintain health recrds fr a perid f 30 years. Recrds must be prvided upn request by representatives f the Chief and/r Directr f NIOSH. Any physician wh cnducts a medical examinatin must furnish the emplyer a statement f the emplyee s ability t safely perfrm the functins f their psitin. Access t the labratry must be restricted t trained persnnel when wrk is being cnducted n tissue. Persnnel handling tissue must be trained in the fllwing: Nature f CJD Rute f transmissin f CJD Page 143 f 201

144 Specific hazards assciated with handling f the tissue. Prin Decntaminatin Prcedures Cntaminated liquids are disinfected by making 1.0N sdium hydrxide (NaOH) fllwed by autclaving at 132º C fr 4.5 hurs. Alternatively (if an autclave that can reach that temperature isn t available), the 1.0N NaOH-treated liquid can be held at rm temperature fr 24 hurs. Liquids treated in either f these ways can then be pured dwn the sink. Cntaminated surfaces that can withstand the treatment are cleaned with 1.0N NaOH, allwing 5 minutes f cntact time, fllwed by wipe-dwn with 1.0N HCl, then thrugh washing with clear water. Cntaminated surfaces that cannt withstand NaOH/HCl treatment are cleaned with 10% husehld bleach, allwing minutes cntact time, then washed with clear water. Cntaminated skin surfaces are washed with 1.0N NaOH r 10% bleach fr 2-3 minutes, fllwed by rinsing with cpius amunts f water. Splashes t the eye are rinsed with cpius amunts f water r saline. Cntaminated dry waste is autclaved at 132º C fr 4.5 hurs then discarded as slid waste (trash). Sharps waste is autclaved at 132º C fr 4.5 hurs befre being picked up as medical waste fr incineratin. Nte: Autclaving f sharps is generally nt recmmended, but in this case, it is recmmended t minimize r eliminate the ptential fr the agent t be intrduced int the envirnment Page 144 f 201

145 Appendix K Summary f Requirements fr Bisafety Levels Safety Guideline Labratry persnnel must wash their hands after handling cultures, remving glves, and befre leaving the labratry. BSL1 BSL2 BSL3 BSL4 Y Y Y Y Eating, drinking, and applicatin f csmetics is prhibited. Y Y Y Y Persnnel must be familiar with basic bisafety prcedures, including this manual. Persnnel shuld wear gggles r face shields if the pssibility f splashes and aersls exists. Y Y Y Y Y Y Y Y Pipetting by muth is prhibited. Y Y Y Y All labratry prcedures shuld be perfrmed t minimize aersl generatin. Wrk surfaces must be decntaminated at least daily, after each use fr infrequent users, and after any spill f viable materials. Sharps must be placed in specially designed puncture- and leak-prf sharps cntainers and dispsed f apprpriately as medical waste. Labratries must be kept neat; gd husekeeping prcedures must be in place and in regular use. All medical waste is decntaminated befre dispsal by an apprved decntaminatin methd r dispsed f as medical waste. Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Insect and rdent cntrl prgrams are instituted. Y Y Y Y Labratry cntains a sink fr handwashing. Y Y Y Y Labratries are designed fr ease f decntaminatin (e.g., n carpets, sealed surfaces, n unreachable areas, etc.). Y Y Y Y Bench tps are impervius t water, mderate heat, and chemicals. Y Y Y Y Labratry furniture must be secured, and spaces between benches, cabinets, and equipment must be accessible fr decntaminatin. Y Y Y Y All labratry windws must be fitted with fly screens. Y Y Y Y Labratry cats r gwns and glves must be wrn. Y Y Y Y Page 145 f 201

146 Safety Guideline Autclaves are required fr waste treatment prir t dispsal as nnbihazardus waste. BSL1 BSL2 BSL3 BSL4 N N Y Y Autclave quality cntrl prgram is required fr use specified abve. Y Y Y Y Instructins fr safety precautins are psted by the Principal Investigatr. Y Y Y Y Animals nt invlved in the experiment are nt permitted in labratry. N Y Y Y Bilgical safety cabinets are required and must be certified annually. N Y Y Y Labratry persnnel require specific training in the handling f pathgenic materials. N Y Y Y Safety centrifuge cups are required. N Y Y Y Access t facility is limited r restricted during experiments. N Y Y Y The universal bihazard symbl must be psted n the access dr t the labratry. Immunizatin and/r serlgical testing fr agents t be handled may be required. All labratry prcedures must be perfrmed in a prperly certified bilgical safety cabinet. Labratry requires cntrlled entry, unidirectinal air flw, and ther special design features. N Y Y Y N Y Y Y N N Y Y N N Y Y Windws must be clsed and sealed. N N Y Y N material r equipment can leave the labratry unless it is autclaved r decntaminated. N N Y Y Autclaves must be lcated inside the labratry. N N Y Y Access is thrugh an airlck system. N N N Y Page 146 f 201

147 Appendix L Summary f Requirements fr Animal Bisafety Levels Safety Guideline Access is limited r restricted at the discretin f the labratry r Animal Care Facility Directr. Persnnel must wash their hands after handling cultures and animals, remving glves, and befre leaving the facility. BSL1 BSL2 BSL3 BSL4 Y Y Y Y Y Y Y Y Eating, drinking, and applicatin f csmetics is prhibited. Y Y Y Y Persnnel must be familiar with basic bisafety prcedures, including this manual. Persnnel shuld wear gggles r face shields if the pssibility f splashes and aersls exists. Y Y Y Y Y Y Y Y Pipetting by muth is prhibited. Y Y Y Y All prcedures shuld be perfrmed t minimize aersl generatin. Y Y Y Y Wrk surfaces must be decntaminated at least daily, after each use fr infrequent users, and after any spill f viable materials. Sharps must be placed in specially designed puncture- and leak-prf sharps cntainers and dispsed f apprpriately as medical waste. Facilities must be kept neat; gd husekeeping prcedures must be in place and in regular use. All medical waste is decntaminated befre dispsal by an apprved decntaminatin methd r dispsed f as medical waste. Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Insect and rdent cntrl prgrams are instituted. Y Y Y Y Drs t animal rms are kept clsed when experimental animals are present. Facilities are designed fr ease f decntaminatin (e.g., n carpets, sealed surfaces, n unreachable areas, etc.). Bedding materials frm animal cages are remved in a manner that minimizes aersl prductin and are dispsed f as medical waste. Y Y Y Y Y Y Y Y Y Y Y Y Instructins fr safety precautins are psted by the Principal Investigatr. Y Y Y Y Facility windws that pen must be fitted with fly screens. Y Y Y Y Page 147 f 201

148 Safety Guideline BSL1 BSL2 BSL3 BSL4 Labratry cats r gwns and glves must be wrn. Y Y Y Y Autclaves are required fr waste treatment prir t dispsal as nnbihazardus waste. Y Y Y Y Autclave quality cntrl prgram is required fr use specified abve. Y Y Y Y Cages are washed prir t release r reuse. Y Y Y Y Air is exhausted t the utside withut recirculatin. N Y Y Y Persnnel baseline serum samples may be required. N Y Y Y Facility persnnel require specific training in the handling f pathgenic materials. N Y Y Y Safety centrifuge cups are required. N Y Y Y Access t facility is limited r restricted during experiments. N Y Y Y The universal bihazard symbl must be psted n the access dr t the facility. Immunizatin and/r serlgical testing fr agents t be handled may be required. All prcedures must be perfrmed in a prperly certified bilgical safety cabinet. Facility requires cntrlled entry, unidirectinal air flw, and ther special design features. N Y Y Y N Y Y Y N N Y Y N N Y Y Windws must be clsed and sealed. N N Y Y N material r equipment can leave the facility unless it is autclaved r decntaminated. N N Y Y Autclaves must be lcated inside the facility. N N Y Y Access is thrugh an airlck system. N N N Y Page 148 f 201

149 Appendix M Bldbrne Pathgen Standard Hepatitis B viral infectin is ne f the mst frequent labratry-assciated infectins, and labratry persnnel are recgnized as a high-risk grup fr acquiring this infectin (Centers fr Disease Cntrl and Preventin). Aviding ccupatinal expsure t human bld, bdy fluids, and tissues is the primary way t prevent transmissin f bldbrne pathgens. The gal f the initial and annual standard precautins training is t present infrmatin n hw t prevent such expsures by administrative cntrls, wrkplace engineering cntrls, prper wrk practices, persnal prtective equipment, and a hepatitis B vaccine immunizatin prgram. Persnnel can be expsed t bldbrne pathgens by being stuck with cntaminated needles, laceratins frm cntaminated sharp instruments, r being splashed with bld r bdy fluids n the mucus membrane f the eye, nse r muth, r n abraded, nn-intact skin (i.e., chapped skin r skin affected by dermatitis). Any direct cntact (i.e., cntact withut barrier prtectin) t cncentrated hepatitis B, hepatitis C, HIV, r any ther infectius virus in a research labratry r prductin facility is cnsidered an expsure that requires clinical evaluatin. All emplyees wrking with human cell cultures shuld be ffered hepatitis B vaccinatin and be evaluated if an expsure ccurs. The OSHA Bldbrne Pathgens Standard applies t all emplyees wh might cme int cntact with bld r ther bdily fluids, including: Human bld Human bld cmpnents Prducts made frm human bld, r ther ptentially infectius materials (OPIM) such as the fllwing human bdy fluids: Semen Vaginal secretins Cerebrspinal fluid Synvial fluid Peritneal fluid Amnitic fluid Saliva in dental prcedures Bdy fluid that is visibly cntaminated with bld, and all bdy fluids in situatins where it is difficult r impssible t differentiate between bdy fluids. Page 149 f 201

150 Any unfixed tissue r rgan (ther than intact skin) frm human (living r dead) HIV-cntaining cell r tissue cultures, rgan cultures, and HIV-, HBV- r HCV-cntaining culture medium r ther slutins; and bld, rgans, r ther tissues frm experimental animals with HIV, HBV r HCV. Prgram Elements The Bldbrne Pathgens Standard requires that an Expsure Cntrl Plan be written and implemented. The Expsure Cntrl Plan must have the fllwing elements: Plicies and prcedures fr eliminatin, r minimizatin, f expsure Evaluatin f emplyee expsure ptentials Medical surveillance prgram Rutine training The fllwing is a general utline f the BU Expsure Cntrl Plan. Rles and Respnsibilities The PI/labratry directr/emplyee supervisr must identify emplyees under his r her supervisin wh may be at risk. Upn identifying these emplyees, the supervisr must Reduce ptential risk by prviding persnal prtective clthing and equipment. Prvide HBV vaccinatins at n cst t the emplyee. Train the emplyees. Develp an effective hazard cmmunicatin prgram. Ensure engineering cntrls, such as a bilgical safety cabinet r sharps cntainer. Develp safe wrk practices and prcedures, as well as internal ntificatin prcedures t reprt accidents. In cnjunctin with the academic department, review the list f at-risk emplyees n an annual basis t ensure that the list is current. Page 150 f 201

151 Key Definitins Other ptentially infectius materials (OPIM) are thse listed in Appendix M. Regulated waste means liquid r semi-liquid bld r ther ptentially infectius materials and cntaminated items that wuld release bld r ther ptentially infectius material in a liquid r semi-liquid state if cmpressed; items that are caked with dried bld r ther ptentially infectius materials and are capable f releasing these materials during handling; cntaminated sharps; and pathlgical and micrbilgical wastes cntaining bld r ther ptentially infectius materials. Sharps waste means any device having acute rigid crners, edges, r prtuberances capable f cutting r piercing, including, but nt limited t, all f the fllwing: Hypdermic needles, syringes, blades, and needles with attached tubing Brken glass items, such as Pasteur pipettes and bld vials cntaminated with ther medical waste At-Risk Emplyee Identificatin The Expsure Cntrl Plan requires each emplyer t identify, in writing, all tasks, prcedures, and jb classificatins where ccupatinal expsure t bld may ccur and t dcument the methds f cmpliance that will minimize the ptential f ccupatinal expsure. Incident Reprting All incidents must be dcumented and a cpy be kept in the labratry and a cpy frwarded t EHS. Accident respnse prcedures are described in Chapter 8. Written Plicies and Prcedures This manual is intended t act as a primary surce f plicies and prcedures designed t eliminate, r minimize, ptential emplyee expsure t all bilgical materials, regardless f their hazard level. Emplyees are required t read and implement all sectins f this manual that are relevant t their wrk envirnment, and be fully familiar with universal precautins. Each PI must further develp site-specific SOP t address lcal prgrammatic needs. Medical Surveillance The OSHA Bldbrne Pathgens Standard requires that all persnnel with ptential expsure t bldbrne pathgens be ffered immunizatin against the hepatitis B virus. HBV vaccinatins must be ffered t an emplyee within 10 days f assignment. Persnnel must indicate their cnsent r declinatin fr the Hepatitis B Vaccine using the ROHP Cnsent r Declinatin frms.. Bth frms are available n the ROHP website at Page 151 f 201

152 The ROHP must retain this frm n file fr the duratin f the emplyee s emplyment plus thirty (30) years. An emplyee wh declines hepatitis B vaccinatin may, at any time thereafter, change his r her mind and receive the vaccine. The acceptance statement must be signed at that time. The PI/labratry supervisr must nt make participatin in a prescreening prgram a prerequisite fr receiving the vaccinatin. The HBV vaccinatin is available at n cst t the emplyee. BPHC must als be ntified f all presumptive expsures (See Appendix U fr BPHC Medical Surveillance Reprting requirements). Training Requirements The PI must ensure that all emplyees with the ptential fr ccupatinal expsure participate in a training prgram prvided by EHS at n cst t the emplyee during wrking hurs. Training must be given in accrdance with the Bldbrne Pathgens Standard upn initial assignment, n an annual basis thereafter, r whenever mdificatin f an existing jb descriptin may affect the emplyee s ptential fr ccupatinal expsure. HIV/HBV research labratries must ensure that their emplyees demnstrate prficiency in standard micrbilgical prcedures prir t being allwed t wrk in the labratry. Training must include a cmprehensive discussin f this standard, including epidemilgy, symptms and transmissin f bldbrne diseases; the Expsure Cntrl Plan; the uses, limitatins f, and prcedures fr using persnal prtective equipment; a discussin f the HBV vaccinatin (including the benefits f vaccinatin and efficiency f the vaccine t prevent disease); emergency prcedures invlving bld expsure r cntaminatin and pst-expsure fllw-up prcedures; hazard cmmunicatin; and a questin-and-answer discussin pprtunity. EHS prvides Bldbrne Pathgens training n a regularly scheduled basis. Fr mre infrmatin and scheduling, call (617) PI Respnsibilities fr Occupatinal Health Issues In keeping with the OSHA Bldbrne Pathgen Standard, this plicy requires annual standard precautins training, a hepatitis B immunizatin prgram, and a pst-expsure medical management prgram. It is the PI s respnsibility t ensure that researchers, technicians, students, r vlunteers wh wrk in the labratry and wh have cntact with animals, infectius agents, r bldbrne pathgens are medically evaluated prir t starting wrk and that anyne wrking with bldbrne pathgens is ffered the hepatitis B vaccinatin series administered by Research Occupatinal Health Prgram in cmpliance with the Bldbrne Pathgen Expsure Plicy fr Bstn University/Bstn Medical Center. PI s are required Page 152 f 201

153 t cmplete the Hepatitis B Vaccine Authrizatin available n ROHP s website at An appintment can be made with a medical prvider by calling BU s Research Occupatinal Health Prgram at (617) (ROHP). If the research prject is lcated at a facility at anther institutin, such as the VA Hspital r the Framingham Heart Study, ccupatinal health services shuld be available n-site at that institutin. Cntact Research Occupatinal Health Prgram at (617) fr assistance in arranging access t apprpriate services. It is the PI s respnsibility t ensure that: Any persn present in a BU labratry wh has an incident invlving ptential expsure t an infectius agent is ffered immediate access t a medical evaluatin at the Research Occupatinal Health Prgram (listed belw) r the BMC Emergency Department (after hurs, hlidays, and weekends). An immediate evaluatin is imprtant, as efficacy f pst-expsure medicatin fr HIV and ther infectius agents may be less effective if the initiatin f treatment is delayed. Persnnel wrking with nn-human primates r their tissues underg an initial health evaluatin including tuberculsis symptm screen and testing. Thereafter, symptm screens and tuberculsis testing are perfrmed every 6 mnths.. Additinally, such individuals are infrmed abut the risks and preventive services available prir t cmmencing wrk with these animals, and wear apprpriate persnal prtective equipment. Persnnel whse jb requires the use f a respiratr must cmplete an OSHA Respiratr Questinnaire fr medical clearance fr respiratr use prir t fit-testing fr a respiratr. Cmpletin f the questinnaire and fit-testing is an annual requirement. Persnnel wh develp symptms f allergy r asthma that ccur upn expsure t experimental animals are referred t the Research Occupatinal Health Prgram fr evaluatin. Under the OSHA Bldbrne Pathgen Standard, BU and BMC are required t ffer the hepatitis B vaccine t all emplyees at risk within 10 days f starting their wrk assignment. Emplyees must be infrmed f the vaccine s benefits and risks, and if they chse nt t receive it at the initial evaluatin time, they must sign a declinatin frm. If the emplyee has had the vaccine previusly, but has nt had a bld antibdy titer t cnfirm his r her immunity in the past, the emplyee will be ffered the pprtunity t have a titer drawn. An emplyee wh declines the vaccine may at any time elect t have the vaccine if his r her jb tasks r wrk setting cntinue t have the risk f ptential expsure t bldbrne pathgens. If at any time, any emplyee has an expsure t bldbrne pathgens, he r she MUST immediately cntact the Research Occupatinal Health Prgram at (617) (ROHP). An immediate evaluatin is imprtant, as efficacy f pst-expsure medicatin fr HIV may be less effective if the initiatin f treatment is delayed. Fr mre infrmatin, call BU s Research Occupatinal Health Prgram at (617) Page 153 f 201

154 Cntacting the Research Occupatinal Health Prgram (ROHP) ROHP is lcated n the Bstn University Medical Campus at 72 East Cncrd Street, 8 th Flr, Rm 825. Our nrmal hurs f peratin are Mnday thrugh Friday frm 8:00am t 4:30pm. Our phne number is (617) (ROHP) and is available and supprted by medical staff 24 hurs per day / 7 days per week t triage and evaluate labratry expsures and related illnesses. Based n injury severity, lcatin and time f day, ROHP will refer peple t the apprpriate health care lcatin using the table belw. 1. Fr lab expsures (needlestick, bite, cut, scratch, splash, etc ) invlving animals r infectius agents n the Medical Campus r Charles River Campus, call the ROHP 24/7 hur number ( ROHP (7647); r, 4-ROHP (7647) if calling frm a Medical Campus lcatin) t be cnnected with the BU Research Occupatinal Health Prgram (ROHP) medical fficer. 2. Fr unexplained symptms r illness call the ROHP 24/7 hur number ( ROHP (7647); r, 4-ROHP (7647) if calling frm a Medical Campus lcatin) t be cnnected with the BU Research Occupatinal Health Prgram (ROHP) medical fficer. ROHP HEALTHCARE REFERRAL LOCATIONS AND TIMES Lcatin Regular Wrkday Hurs After Hurs and Weekends Medical Campus BU ROHP 72 East Cncrd St., 8 th Flr, Rm BMC OEM 850 Harrisn Ave Yawkey ACC BMC Emergency Department 751 Albany Street (Mn-Fri 8:00a.m. t 4:30p.m.) (Mn-Fri 7:30a.m. t 4:00p.m.) (Reminder: Present Agent Card) Charles River Campus Hme r Away Frm Wrk BU Occupatinal Health Center (OHC) 930 Cmmnwealth Avenue, West (Mn-Fri 9:00a.m. t 5:00p.m.) Nt Applicable BMC Emergency Department 751 Albany Street (Reminder: Present Agent Card) Primary Care Physician r Lcal Hspital (Reminder: Present Agent Card) 3. Under any f these scenaris, always infrm the physician f yur wrk in the labratry and the agent(s) that yu wrk with. 4. If yu have been given a wallet-size agent ID card, prvide the agent ID card t the physician. Any questins may be directed t: Research Occupatinal Health Prgram at (617) (ROHP). Page 154 f 201

155 REFERENCES: OSHA Bldbrne Pathgen Prgram: Centers fr Disease Cntrl and Preventin/Natinal Institutes f Health: Bisafety in Micrbilgical and Bimedical Labratries, U.S. Department f Health and Human Services, 5 th editin, February CDC: Updated U.S. Public Health Service Guidelines fr the Management f Occupatinal Expsures t HBV, HCV, and HIV and Recmmendatins fr Pst-expsure Prphylaxis, Mrbidity and Mrtality Weekly Reprt, June 29, 2001/Vl. 50/ N. RR Institute f Labratry Research (ILAR): Occupatinal Health in the Care and Use f Research Animals, Natinal Academy Press, U.S. Department f Labr/OSHA: Bldbrne Pathgens and Needlestick Preventin. Page 155 f 201

156 Appendix N Wrking Safely with Animals Wrking with animals pses ptential additinal health and safety hazards that require extra precautins. The specific requirements will depend n the types f activities (e.g., surgery, feeding animals, use f anesthetic agents, etc) and the specific species used. Persnnel shuld fllw the fllwing guidelines: Fllw the specific requirements established in the IACUC-apprved prtcl and the facility requirements. Wash hands after handling an animal r anything that an animal has tuched. The mst cmmn way f cntracting an animal-transmitted infectin is placing the infectius material directly int the muth. Never smke, drink, r eat in an animal area r befre washing hands. Wear prtective clthing as recmmended by the facility fr the species and peratins. D nt take prtective clthing hme. Prtective clthing helps prevent ptentially cntaminated material frm leaving an animal area. Use the persnal prtective equipment (PPE) recmmended fr the species and peratins. Wrkers shall wear the apprpriate PPE (e.g., glves, face shields, masks, and respiratrs) when required and fllw their supervisr s instructins scrupulusly. Participate in the Research Occupatinal Health Prgram medical surveillance prgram (See Appendix R Medical Surveillance Prgram) that prvides medical evaluatins, testing, immunizatins, and peridic screenings fr allergies, infectins, and ther medical prblems related t animal expsure. Seek medical attentin prmptly when injured. Fllw the specific recmmendatins fr the facility. Wrkers engaged in wrk invlving vertebrate animals shuld infrm their physician f their wrk when seeking treatment fr illness, even if uncertain whether the illness is wrk related. All animal care wrkers are prvided wallet-sized agent identificatin cards. The card indicates the card carrier wrks in a labratry setting at Bstn University and may be expsed t hazardus materials. The card als cntains ROHP cntact infrmatin in the event the physician shuld chse t seek further infrmatin n ptential ccupatinal expsure. Physicians need such infrmatin t make an accurate diagnsis because many animaltransmitted diseases have flu-like symptms. Page 156 f 201

157 If there is any pssibility f wrk-related illness r disease, the Research Occupatinal Health Prgram must be ntified immediately at (617) (ROHP). Get the apprpriate training and cntact a supervisr with any questins. Basic Safety fr the Necrpsy f Infected Animals Ensure that the necrpsy f infected animals is carried ut in bilgical safety cabinets by trained persnnel. Wear a surgen s wrap-arund gwns ver labratry clthing. Use a surgen s mask and eye prtectin. Use ther PPE recmmended by the facility fr the infectius agents present. Wear glves. Wet the fur f the animal with a suitable disinfectant. Pin dwn r therwise fasten small animals t metal in a tray. Befre and after necrpsy, disinfect the necrpsy table, inside the BSC, and ther ptentially cntaminated surfaces with a suitable germicide. Upn cmpletin f necrpsy, place all ptential bihazardus materials in suitable cntainers and then sterilize the materials. Segregate cntaminated mixed waste and stre fr apprpriate dispsal. Place cntaminated instruments in a bath that cntains a suitable disinfectant. Fllw the facility requirements fr sterilizatin. Clean cntaminated rubber glves in disinfectant befre remval frm the hands. Wearing glves is nt a substitute fr handwashing; wash hands after necrpsy and carcass dispsal. Fllw the facility s guidelines fr the dispsal f dead animals Page 157 f 201

158 Appendix O Prcedures fr Wrking in an Animal Bisafety Level 2 (ABSL-2) Facility at BU Befre starting any Animal Bisafety Level 2 (ABSL-2) wrk at BU/BMC, a PI must: Obtain IBC and IACUC apprval Make apprpriate husing arrangements with the LASC/LACF directr The fllwing Standard Operating Prcedures (SOP) have been develped t prvide guidance t thse individuals wrking in rms in which animals invlved in chemical and bilgical hazards determined t be ABSL-2 are hused. Definitins Overview ABSL-2: Animal Bisafety Level 2 includes pathgenic agents f mderate hazard ptential (CDC Bihazard Class 2) and chemical hazard agents f mderate hazard ptential. PPE: Persnal prtective equipment EHS: Envirnmental Health and Safety Parenteral: Taken int the bdy r administered in a manner ther than thrugh the digestive tract, as by intravenus r intramuscular injectin. Access t the rm where the wrk with animals is t be cnducted is restricted. Labratry persnnel must have training in aseptic micr-islatr techniques, when applicable, and use f bilgical safety cabinets, in additin t specific safety training in handling the pathgenic and/r chemical agent(s) with which they are wrking. Research and Labratry Animal Science Center (LASC at BUMC) and Labratry Animal Care Facility (LACF at CRC) persnnel shuld receive apprpriate immunizatins r tests fr any agents handled r ptentially present in the rm prir t initiating the ABSL-2 prtin f their prject. Prcedures must be cnducted in a Class II BSC. Persnal Prtective Equipment Minimum PPE: Slid frnt gwn: Page 158 f 201

159 Hair cver She cvers Mask In additin: Duble glves N95 may be required Face shield r ther specific eye r face prtectin may be required Equipment and Supplies Bihazard stickers fr cage cards MB-10 (chlrine dixide) disinfectant r Virkn-S Bilgical safety cabinet: Class II Respnsibilities It is EHS respnsibility t ensure that all necessary prject-specific safety training is prvided t research and LASC/LACF staff prir t any prject being initiated. It is als EHS respnsibility t prvide dcumentatin t the LASC/LACF f such training. The PI and individuals wrking in the facility are respnsible fr ensuring they have received prper training and that they are adhering t this SOP, as well as t psted precautins and guidelines in the facilities. Prcedure Entry Remve the lab cat wrn in the LASC/LACF facility and hang it n the garment rack prvided utside f designated ABSL-2 animal space. PPE must be wrn while wrking in ABSL-2 animal husing and prcedure space. PPE is prvided just utside specific ABSL-2 rms. Dn designated PPE prir t entry int the ABSL-2 areas. Prceed int the designated ABSL-2 rm using an access card r key. Cnducting a Prcedure General Infrmatin Page 159 f 201

160 Investigatrs using the rm will be assigned a cubicle and/r rack and shelf/shelves where their animals will be hused. A cubicle r rack may hld cages belnging t mre than ne investigatr. Bihazard prjects are nt hused in cubicles that huse nging chemical hazard prjects. LASC/LACF persnnel will perfrm daily health checks f animals n studies invlving infectius agents if the animals are hused n racks nt held in cubicles. LASC/LACF persnnel will perfrm daily health checks f animals n studies invlving chemical agents by viewing the animals thrugh the windw f the islatin cubicle. The LASC/LACF will ntify PIs f any animal health issues. All animal wrk will be cnducted within the cnfines f the Class II BSC. Always pen animal cages in the Class II BSC using aseptic micrislatr technique. Hypdermic needles and syringes are used nly fr parenteral injectin r aspiratin f fluids frm labratry animals and bttles with plastic/rubber diaphragms. Only needle-lcking syringes r dispsable needle syringe units (i.e., the needle is integral t the syringe) are used fr the injectin r aspiratin f infectius fluids. Needles shuld nt be bent, sheared, replaced in the sheath r guard, r remved frm the syringe fllwing use. The needle and syringe shuld be prmptly placed in a puncture-resistant sharps cntainer. Always spray r wipe dwn all interir surfaces f the Class II BSC with MB-10 r Virkn-S befre and after wrking in the hd. Allw 10 minutes f cntact time prir t wiping the surfaces with dispsable twels. D nt spray the tp grille f the Class II BSC (this is where the filter is lcated). Discard used twels in the waste cntainer. Page 160 f 201

161 D Nt spray MB-10 r ther disinfectants here. Immediately fllwing infectin f animals with pathgenic agents, place a bihazard sticker n their cage card(s) (LASC/LACF prvides these specific stickers). Fill ut the fllwing infrmatin n the bihazard sticker fr cage cards: PI name 24-hur cntact phne number Prtcl number Pathgenic agent Dse per animal Date(s) infected Husbandry by PI r LASC/LACF (circle ne) Reprt all spills and accidents that result in vert expsure t infectius materials t a LASC Supervisr and the LASC ffice, (617) r LACF at (617) Remval f Dirty Cages and Bttles frm Rm Bilgical agents Place a cage inside the BSC, remve the water bttle frm the cage, and replace the lid. Page 161 f 201

162 Put siled cage, wire lid, and bedding in a semi-clear bihazard autclave bag and place n a cart. Put water bttles in a separate, semi-clear bihazard autclave bag n a cart. Clsure ties are stred adjacent t the bihazard autclave bags n the supply rack in the crridr utside rms and in rms. D nt use autclave tape t clse the bag. Thrughly spray the exterir f all bags with MB-10 r Virkn-S after clsing the bag with a nyln tie, beaded tie, r twist tie. Spray all surfaces and wheels f the cart(s). Cart(s) shuld be mved t the siled side f the cage wash rm. Chemical agents Place a cage inside the BSC, remve the water bttle frm the cage, and replace the lid. Put siled cage, wire lid, and bedding in a red bihazard bag and place n a cart. Put water bttles in a separate red bihazard bag n a cart. Clsure ties are stred adjacent t the bihazard autclave bags n the supply rack in the crridr utside W-838/839 and in bth rms. D nt use autclave tape t clse the bag. Thrughly spray the exterir f all bags with MB-10 after clsing the bag with a nyln tie, beaded tie, r twist tie. Spray all surfaces and wheels f the cart(s). Cart(s) shuld be mved t the siled side f the cage wash rm n the 8 th flr f W Building (W-8). Dispsal f carcasses Any dead animals must be remved frm their cage (while in the BSC) and placed in a small, leak-prf red bihazard bag. Place a sticker with animal identificatin infrmatin n the uter bag, then thrughly spray the bag with MB-10 r Virkn-S and place in the refrigeratr. A cage card with a sticker shwing the same animal identificatin infrmatin will be placed n the cage. Page 162 f 201

163 The LASC/LACF will remve carcasses fr incineratin three days after they are fund in the refrigeratr. Exiting Prcedures When wrk is cmplete and the BSC and all ther wrk spaces have been decntaminated with MB-10 r Virkn-S, uter glves shuld be remved and placed in the bihazard waste cntainer. T exit the rm, pen the dr and remve ne she cver, stepping ver the rm threshld int the hallway with that ft. Remve the she cver frm the ther ft as it is brught int the hallway but befre stepping int the hallway with the secnd ft. Remve gwn frm the shulders, turning it inside ut. Discard dispsable face prtectin, hair cver, mask, gwn and glves in the red bihazard trash receptacle in the hallway utside ABSL-2 rm. After leaving ABSL-2 rm and discarding PPE, hands shuld be washed using the alchl hand sprayer lcated n the wall immediately t the left f the drs t each ABSL-2 rm Reference: Bisafety in Micrbilgical and Bimedical Labratries (5 th editin, 2007) Page 163 f 201

164 Appendix P Plicy fr Verifying the Identity f Attenuated Pathgens This plicy cvers all attenuated pathgens that meet the fllwing criteria: 1. The attenuated pathgen is derived frm a knwn, virulent pathgen that requires BSL3 r BSL4 bisafety cntainment; 2. Attenuatin results in decreased virulence f the rganism; 3. Attenuatin results in a reductin in the level f bisafety cntainment in which the attenuated pathgen can be handled, cmpared with the bisafety level required fr the safe handling f the nnattenuated cunterpart. This plicy des nt apply t pathgens that have been inactivated by a prcess established and accepted as scientific and safety standard and apprved by the IBC. The necessary requirements t wrk with inactivated pathgens derived frm BSL3 r BSL4 cntainment facilities are cvered in the Validatin f Inactivated Pathgens plicy. The Inactivated Bilgical Samples Use Frm may be fund n the IBC website at: As is the case with all prpsed studies at Bstn University and Bstn Medical Center, wrk with any pathgen, including attenuated and inactivated pathgens, cannt be initiated (and the pathgens cannt be brught int BU) withut prir apprval frm the Institutinal Bisafety Cmmittee (IBC). Plicy The identity f any attenuated pathgen must be verified befre it can be handled at the lwer bisafety cntainment level f the attenuated pathgen. Verificatin shuld be by restrictin analysis r anther methd that has been prven t rigrusly distinguish the wild-type and attenuated pathgens. Verificatin shuld be cnducted n the received stck and, whenever pssible, verificatin shuld be carried n an individual clne f the incming attenuated pathgen. Only material frm the verified clne (r stck) can be studied at the lwer bisafety level. Verificatin shuld be carried ut by the receiving BU labratry pursuant t an IBC-apprved prtcl fr verificatin. The attenuated pathgen must nt be received at BU until after the IBC apprval f the prtcl fr verificatin, including hw and where the specimen will be stred upn receipt. Upn receipt f the specimen, the pathgen must be handled at the higher bisafety level f the nn-attenuated, wildtype pathgen until the specimen has been verified t cntain nly the attenuated pathgen that can be Page 164 f 201

165 safely handled at the lwer bisafety level. Verificatin must be submitted t the IBC, r their designee, fr review prir t transferring the attenuated pathgen t the lwer cntainment level. If the identity f an attenuated pathgen cannt be verified, it must be handled at the higher bisafety level f the nn-attenuated wild-type pathgen. Fd and Drug Administratin (FDA)-apprved human vaccines that cntain attenuated BSL2 agents derived frm BSL3 r BSL4 agents may be excluded frm the BU strain verificatin plicy if the frmulated vaccines are btained directly frm the vaccine manufacturer. EHS and IBC ffices will review the manufacturer s dcumentatin t determine whether exclusin frm the BU plicy is warranted and can request further review by the full IBC. The IBC may cnsider and apprve strain verificatin dcumentatin prvided by ff campus prducers f attenuated BSL2 strains derived frm BSL3 r BSL4 pathgens in certain instances. Strain verificatin must be recent and prvidence must be evident, dcumenting the strain handling frm verificatin thrugh shipment t Bstn University and Bstn Medical Center. This dcumentatin will be reviewed by EHS and IBC ffices which can request further review by the IBC. The Strain Verificatin Prcedure with Template Frm may be fund n the IBC website at: Page 165 f 201

166 Appendix Q Bstn Public Health Cmmissin Requirements The Bilgical Labratry Regulatins adpted by the Bstn Public Health Cmmissin (BPHC) in September 2006 require labratries at Bisafety Level 3 (BSL-3) and BSL-4 t adhere t new lcal standards, including: Obtaining a permit frm the Bstn Public Health Cmmissin. Setting up an Institutinal Bisafety Cmmittee (IBC) that reprts t a senir respnsible fficial and that has at least tw cmmunity members withut a cnnectin t the rganizatin. Submitting reprts t the BPHC n research, safety prcedures, and IBC meetings. Fllwing the highest safety standards utlined in the CDC/NIH manual n bisafety, Bisafety in Micrbilgical and Bimedical Labratries. Reprting labratry incidents r illness, and inspectins. Prhibiting weapnizatin and classified research in Bstn. Hlding annual public meeting t review the type and nature f the bilgical research. Reprting intended decmmissining f a labratry facility. Fllwing rdna r medical surveillance regulatin requirements. Establishing a Cmmunity Benefits Prgram t supprt lcal health and safety needs as a mandate fr BSL-4 labratries. Page 166 f 201

167 Appendix R ROHP Medical Surveillance Prgram BU and BMC prvide free medical mnitring t all emplyees wh face wrkplace risks. The prgram is designed t mnitr ptential health hazards assciated with research and develpment activity with recmbinant DNA, bldbrne pathgens, ther etilgic agents, zntic diseases assciated with labratry animals, and hazardus chemicals. The details f the prgram are prvided belw: I. Objectives: The Medical Surveillance Prgram develped by the Research Occupatinal Health Prgram has the fllwing bjectives: a. Determine the initial and peridic medical surveillance requirements fr thse persnnel that perfrm research and thse grups that supprt research such as animal care wrkers, EHS, Public Safety, and Facilities (see Appendix A n the ROHP website) b. Define the surveillance requirements based n the wrk envirnment, ccupatinal expsure and risk, and access requirements fr each psitin. c. Determine whether the emplyee r applicant is able t safely perfrm the essential functins f the jb fr which emplyment has been ffered. d. Determine accmmdatins, if necessary, fr an emplyee r applicant t perfrm the functins f the jb in a safe and effective manner. e. Establish a baseline fr cmparisn with future peridic evaluatins and terminatin evaluatins. f. Establish a prcedure fr perfrming additinal medical surveillance in supprt f the Institutinal Bisafety Cmmittee (IBC) when new prtcls are reviewed and changes in the jb functin r rle, expsure t hazardus materials and access requirements fr researchers arises. II. Scpe: The scpe f medical surveillance prvided by ROHP will include the fllwing activities and are prvided t the ppulatin identified belw: a) The clinical services prvided as part f the Initial and Peridic medical surveillance prfiles by jb grup in Appendix A includes questinnaires, physical examinatins, testing, and screenings dictated by the expsure type and access/lcatin requirements fr each psitin: i. Expsure Type: agents, animal types, lasers, chemical r ther hazardus materials, blds, tissues, cells r fluids, r patient care ii. Access/Lcatin: Research Labratry and Bisafety Level, NEIDL, and Animal Care Facilities Page 167 f 201

168 b) In additin t the clinical services identified abve, bilgical agent immunizatin requirements are als defined based n the specific agents individuals wrk with r are ptentially expsed t. The Initial and Peridic Bilgical Agent Specific Immunizatins are identified in Appendix B (see ROHP website). c) These clinical services will be prvided t the emplyees r applicants wh perfrm research, supprt research, r require access t research facilities in the jb grups prvided belw and defined in Appendix A: iii. Grup One: Researchers iv. Grup Tw: Animal Care Wrkers v. Grup Three: Envirnmental, Health and Safety (EHS) vi. Grup Fur: BU Plice vii. Grup Five: Public Safety viii. Grup Six: Facilities and Trades ix. Grup Seven: Other 1. Administrative 2. Telc 3. Infrmatin Technlgy 4. Students, Cntractrs and Temprary Persnnel r Vlunteers a. Fr the NEIDL facility, Public Safety will determine whether these individuals have escrted r unescrted access. b. If unescrted NEIDL access is required, medical surveillance will be perfrmed based n minimum NEIDL requirements plus ccupatinal expsure and risk, and access level required. c. N medical surveillance is required fr escrted NEIDL visitrs. d) An Example f Hw t Use Appendix A and B t Define Initial and Peridic Medical Surveillance Cmpnents Using Sample Researcher is prvided in Appendix C (see ROHP website). This is a step by step example that illustrates hw t use Appendix A and B t determine individual medical surveillance prfiles based n jb grup, expsure types, and access/lcatin requirements. e) Exit Examinatins will be perfrmed fr the fllwing persnnel: Page 168 f 201

169 i. EHS persnnel perfrming emergency respnse whse last peridic examinatin was perfrmed mre than six mnths prir t the date f terminatin ii. Any researcher cleared t wrk in a BSL 3 r BSL 4 research labratry envirnment whse last peridic examinatin was perfrmed mre than six mnths prir t the date f terminatin iii. Any persnnel currently mnitred by ROHP as part f a pst-expsure fllw-up and mnitring plan f) Medical surveillance fr transfers, rehires, r emplyees returning frm leave: i. All transfers int the NEIDL require mental health and drug screens alng with a pre-placement baseline examinatin, testing and immunizatins in accrdance with Appendix A and B. Certain testing and immunizatins may be avided If dcumentatin f previus baseline examinatins and testing is available and within current guidelines. ii. Emplyees returning t Bstn University Research: a. Frm leaves f absence f mre than ne year r previus emplyment at BU mre than ne year ag, a cmplete medical surveillance is required. b. Frm leaves f absence f less than ne year r previus emplyment at BU less than ne year ag, cmpletin f an abbreviated health questinnaire is required. N ther examinatins are needed unless health risks are indicated in the abbreviated questinnaire. iii. Emplyees returning t BU Research that require access t the NEIDL: a. Frm leaves f absence f mre than ne year r previus emplyment at BU mre than ne year ag, a cmplete medical surveillance is required. b. Frm leaves f absence f less than ne year r previus emplyment at BU less than ne year are required t cmplete an abbreviated health questinnaire and underg mental health and drug screens. N ther examinatins are required unless health risks are indicated in the abbreviated questinnaire. III. Prcedures: The prcedures fllwed by ROHP medical persnnel in the determinatin and perfrmance f medical surveillance are as fllws: a) Researchers will cmplete an Initial Health Questinnaire (IHQ) including the ccupatinal expsure and risk assessment cmpleted in Part A. Page 169 f 201

170 b) The nurse in ROHP reviews the IHQ fr ccupatinal expsure and risk assessment, immunizatins and medical limitatins t essential jb functins. c) ROHP cntacts the applicant/emplyee t discuss additinal dcumentatin and testing needed fr medical clearance. d) ROHP schedules an appintment in ROHP fr required examinatin cmpnents as needed. e) If n additinal infrmatin is needed, the nurse in ROHP cmpletes a Medical Clearance Frm and clears the Researcher t begin wrk. A cpy f the Medical Clearance Frm is maintained in the Medical Recrd, and a cpy is given t the applicant/emplyee. f) If a physical examinatin is required, ROHP schedules the exam with an ROHP Healthcare Prvider. g) The Healthcare Prvider reviews all testing results, cmpletes a physical examinatin, and, is available t cnfidentially discuss any health issues with the applicant/emplyee. The Healthcare Prvider cmpletes a Medical Clearance Frm, files the frm in the individual s Medical Recrd and gives a cpy t the applicant/emplyee. h) Fr peple requiring NEIDL access, a mental health screen and drug screen are required fr medical clearance. The Healthcare Prvider reviews the results and is available t cnfidentially discuss any health issues with the applicant/emplyee. The Healthcare Prvider cmpletes a Medical Clearance Frm, files the frm in the individual s Medical Recrd and gives a cpy t the applicant/emplyee. i) The ROHP ntifies the apprpriate department when a Researcher is medically cleared. j) Researchers may be asked t cmplete additinal questinnaires depending n their jb functin, i.e. OSHA Respiratr Users Questinnaire fr respiratr users, Animal Allergy Questinnaire fr Researchers with animal allergies and wrking with animals. k) Annual Health Questinnaire is cmpleted by all persnnel. The annual questinnaire is used t review any new medical cnditins, medicatins, wrk expsures r prcesses that may require additinal medical surveillance s that early preventive strategies can be recmmended. l) Annual Respiratry Questinnaire will be cmpleted by persnnel whse psitin requires the use f any type f respiratr ther than a clth surgical mask. m) Symptm screen surveys will be cmpleted by Researchers every 6 (six) mnths if their research invlves Mycbacterium tuberculsis, r, wrk with r have air expsure t nnhuman primates. This bi-annual survey discvers symptms r cnditins that increase the pssibility f early tuberculsis infectin. Page 170 f 201

171 i. Tuberculsis testing prcedures will fllw the Tuberculsis Screening Prtcl. IV. Respnsibilities: The respnsibilities fr the functinal grups invlved at any level f the ROHP Medical Surveillance Prgram are as fllws: a. Principal Investigatr (PI) r hiring manager: i. Requests a jb requisitin psting frm human resurces fr a new psitin ii. Cmpletes jb specific infrmatin required by human resurces t pst the psitin: b. Human Resurces c. Candidate 1. PS-1 frms 2. Jb Specific Risk Assessment Frm identifying the specific ccupatinal expsure and risks f the wrk envirnment fr the psitin i. Ntifies ROHP t schedule a medical evaluatin fr emplyees r applicants seeking psitins in research r supprting research ii. Prvides the candidate with the Jb Specific Risk Assessment Frm cmpleted by the PI r hiring manager fr this specific psitin. iii. Directs the candidate t the ROHP website t cmplete the Initial Health Questinnaire (IHQ). iv. Fr NEIDL jb applicants, perfrms backgrund check (criminal and credit) in additin t the abve fr Ntifies NEIDL Public Safety whether cleared r nt. v. Fr NEIDL jb applicants, includes infrmatin abut NEIDL medical clearance prcedures in cnditinal ffer f emplyment including drug testing prcess, testing lcatins, and Chain f Custdy frms needed t cmplete the prcess. vi. Crdinates ROHP medical clearance ntificatins with emplyee r applicant, hiring manager and Public Safety. i. Ges t the ROHP website, fr access t the IHQ. The IHQ cnsists f tw parts: 1. Part A: Identifies the ccupatinal expsure and risk assessment fr the psitin t be filled by the candidate. Page 171 f 201

172 d. ROHP 2. Part B: Candidate s medical histry infrmatin and cnsents fr examinatin and authrizatin fr disclsure ii. Cmpletes the IHQ: 1. Candidate incrprates infrmatin frm the Jb Specific Risk Assessment Frm cmpleted by the PI r hiring manager int Part A f the IHQ 2. Candidate prvides medical histry infrmatin in Part B f the IHQ 3. Candidate prvides cnsent fr examinatin and authrizatin fr disclsure in Part B f the IHQ iii. Cmplies with additinal requests fr infrmatin and required testing in a timely manner i. Cntact candidates ( , phne, mail) ii. Request cmpletin and return f Initial Health Questinnaire (IHQ) iii. Reviews the candidate s IHQ. The purpse f the IHQ is t: 1. Define the medical surveillance required based n ccupatinal expsure and risk f the wrk envirnment fr the candidate s psitin. 2. Establish a baseline medical histry fr the candidate fr nging medical surveillance, and 3. Assess the candidate s ability t safely perfrm the functins f the psitin. iv. Determines additinal medical dcumentatin needed, i.e. immunizatin recrds, tuberculsis screens, etc. v. Schedules physical examinatin, additinal testing (labs, pulmnary functin test, electrcardigram, as needed accrding t expsure ptential t agents, risk, cntact (see Appendix A and B n the ROHP website). vi. Schedules mental health and drug screening fr persnnel requiring NEIDL access vii. Reviews results f all testing, screenings and examinatins viii. Ntifies apprpriate persnnel f examinatin utcme Page 172 f 201

173 1. Medically cleared t perfrm essential functins f the jb 2. Medically cleared t perfrm essential functin f the jb with the fllwing restrictins: 3. Examinatin incmplete due t 4. Medically nt cleared t perfrm essential functins f the jb ix. Issues generic Agent Card fr individuals fr persnnel wh may be expsed t hazardus materials while wrking in a research r animal care facility. The card cntains ROHP cntact infrmatin and is used t facilitate prmpt medical attentin and apprpriate medical care in the event the card hlder shuld experience symptms r illness while away frm Bstn University that may be related t activities r expsures in a labratry research envirnment. e. Envirnmental, Health and Safety (EHS) i. EHS will identify thse persnnel with ptential expsure risks that warrant baseline and/r additinal mnitring, i.e. Respiratry Prtectin, Nise, Laser (baseline fr 3b r 4 laser users nly), and Emergency Respnders ii. EHS will cmmunicate similar expsure grup data (names, expsure type) annually t ROHP after discussin with Principal Investigatrs and Labratry Managers, i.e. Nise, Laser. iii. EHS will crdinate training (bisafety level and agent specific) and ptential risk expsure with PI and ROHP iv. EHS will cnduct Annual Respiratr Fit Testing and Respiratry Prtectin Safety Training. v. EHS will cnduct safety training apprpriate t emergency prtcls and general labratry safety issues, such as lck ut/ tag ut, fire safety, etc. vi. Issue Agent Specific Identificatin Cards t all labratry persnnel apprved by the IBC t wrk with bilgical agents with the ptential t cause LAI (Appendix I). This card cntains ROHP cntact infrmatin and is prvided t facilitate prmpt medical attentin and apprpriate medical care in the event the card hlder shuld experience symptms r illness while away frm Bstn University that may be related t activities r expsures in a labratry research envirnment. f. Public Safety i. Ntifies ROHP when an emplyee r applicant has been apprved t enter the NEIDL medical surveillance prcess and prvides ROHP with access level required Page 173 f 201

174 V. Recrdkeeping ii. Prvides emplyee r applicant with NEIDL security access after all clearance cnditins have been met including medical clearance frm ROHP. Updates clearances annually frm security and safety perspective a. Refer t the Recrdkeeping Guideline. b. Medical recrds will be maintained in the ROHP ffices. c. Electrnic medical recrds will als be maintained fr all persnnel seen in ROHP. VI. Attachments (available at ROHP website, a. Initial Health Questinnaire b. Annual Health Questinnaire c. Animal Allergy Questinnaire d. OSHA Respiratr Users Medical Questinnaire e. Tuberculsis Testing and Symptm Screening Questinnaire f. Bstn Public Health Department Tuberculsis Clinic Referral Frm fr Psitive TB Testing Result g. Immunizatin Cnsent Frms h. Vaccine Infrmatin Sheets VII. Appendices (available at ROHP website, i. Appendix A: Initial and Peridic Surveillance Requirements by Grup j. Appendix B: Initial and Peridic Bilgical Agent Specific Immunizatin Requirements k. Appendix C: Example n Hw t Use Appendix A and B t Define Initial and Peridic Medical Surveillance Cmpnents Using Sample Researcher Page 174 f 201

175 Appendix S Labratry and Equipment Decntaminatin Prcedures Decntaminatin f Lab Space and Equipment Punch List 1. Designate a Mve Crdinatr. 2. Cntact an utside vendr fr the decntaminatin f bilgical safety cabinets (tissue culture hds). 3. Have apprpriate persnal prtective equipment available (lab cat, glves, eye prtectin). 4. Dispse f ld chemicals and all ther chemical waste as hazardus waste. Ntify the Envirnmental Manager at (617) at BUMC r (617) at CRC upn terminatin f a hazardus waste accumulatin area r with any questins n this issue. 5. Decntaminate all equipment that is either t be mved r left behind. 6. Cntact EHS regarding the discarding f equipment. 7. On the decn certificate, list all decntaminated equipment by rm (ne sheet per rm). 8. Small pieces f equipment can be decnned and bxed by lab persnnel. 9. Any wrking equipment t be left behind withut a new wner must be reprted t Facilities Management and EHS. 10. Cntact the Radiatin Prtectin Office (RPO) fr the decntaminatin and mving f radilgical materials and wrk spaces. 11. Decntaminate all labs, including fume hds, the utside f tissue culture hds, cld/warm rms, darkrms, etc. 12. If perchlric acid was used in a fume hd, cntact EHS at (617) at BUMC r (617) at CRC. 13. Fill ut ne decn sheet fr each rm, tape ne cpy t the utside f the lab dr (if it is a sectin f a lab, tape t bench), fax ne cpy t the Bisafety Office, and keep ne cpy fr the recrds. 14. Disinfectants: the mst cmmn are 10%freshly diluted bleach (leave n fr minutes, then wash ff), 70% EtOH, r isprpanl. Phenlic agents are nt recmmended. 15. If refrigeratrs, freezers, incubatrs, etc., are t be mved with cntent inside, make sure the cntent is well prtected frm sliding, breaking, etc. Page 175 f 201

176 16. The Mve Crdinatr must ensure the fllwing emergency prcedures are cvered: Chemical spills Bilgical spills Fire Persnal injuries, such as slips, falls, cuts, etc. 17. Prtective clthing and spill absrbent materials must be n hand. 18. Fllw and cmplete the Labratry r Equipment Decntaminatin certificatin frm. EHS will inspect the labratry space r equipment t ensure they have been apprpriate cleaned and decntaminated. 19. EHS will affix a decntaminatin sticker n equipment that had been prperly cleaned and disinfected. The equipment will be mved ut within 15 days that the sticker is issued. The equipment will nt be mved if the 15 days issuance lapses. EHS will inspect the equipment again and issue a new sticker. 20. Bihazard sticker will be remved nce the equipment has been prperly decntaminated. Page 176 f 201

177 LABORATORY OR EQUIPMENT DECONTAMINATION CERTIFICATION Date: Lcatin and Department: 1 I certify that the rms r equipment listed belw, previusly used by my labratry, have been emptied f bilgical and chemical materials: EQUIPMENT: 2. The surfaces f these rms/equipment have been decntaminated (if equipment: inside and utside) with: (specify decntaminants and percentages, (i.e. 70% Ethanl, if 10% bleach is used, it must be freshly made up). 3. All chemicals cntained within the rms r equipment have been remved r drained and cllected fr prper dispsal (including but nt limited t): Oil if the equipment cntains a pump r ther il reservir, il must be drained and cllected as Hazardus Waste in the labratry s Satellite Accumulatin Are. Cntact EHS. Mercury If there is a thermmeter r ther device inside r assciated with the equipment r space the device must be remved and cllected as Hazardus Waste in the labratry s Satellite Accumulatin Area. Cntact EHS fr assistance. Refrigerant Gas If the equipment invlved cling and relied n refrigerant gas, this gas must be remved prir t dispsal. Facilities Management must be cntacted as nly licensed mechanics can perfrm this service. Lead Shielding If the equipment used lead as a shielding agent, this material must be remved prir t dispsal. Cntact EHS t assist in lead remval. Yes N N/A 4. If the space r equipment cntained r was used with any radiactive materials (istpes, sealed surces, etc.), the labratry persnnel have decntaminated the area and equipment. Radiatin Safety has been cntacted, has surveyed the equipment, and has certified it free f detectable Radiactive cntaminatin and arranged fr the remval f any shielding: Page 177 f 201

178 *Cmplete a radilgical Equipment Release Survey Request, available n the radiatin Safety website at: 5. All sink traps (including thse in fume hds0 have been bleached and flushed with water (use 1 cup f cncentrated bleach, wait 20 minutes, then flush thrughly with water): Yes N N/A 6. Des the equipment cntain fluid (e.g. water bath, antifreeze, etc.): Yes N Name (print): Signature: Phne Ext: Date: Dept., Bldg., Rm #: Decntaminated By: Principal Investigatr: Please return/fax cmpleted frm t Envirnmental Health and Safety BMC/BUMC: Fuller Building, 4th Fl (M-470), / CRC: 704 Cmm. Ave., 2nd Fl, Page 178 f 201

179 Labratry Decmmissining and Relcatin Prcedures Purpse and Applicability Definitins It is the plicy f Bstn Medical Center and Bstn University that labratry decmmissining take place prir t the relcatin f any labratry space r upn vacating labratry space r leaving either institutin. In additin, safe-mving practices must be adhered t at all times. This plicy is intended t minimize research and clinical lab dwntime due t mving f a labratry, and t prtect cntractrs, labratry persnnel, and any ther persnnel invlved in the prcess frm labratry hazards. This plicy applies t all Bstn Medical Center and Bstn University emplyees and tenants ccupying labratry space within Bstn Medical Center and Bstn University buildings. Abandned Labratry: A clinical r research labratry that is left vacant by a Principal Investigatr r Labratry Directr and his r her labratry staff, and has labratry materials (bilgical, surplus chemical, radiactive), equipment r waste that has nt been dispsed f. Bilgical Materials: All human, plant, and animal pathgens; all human bld, bld cmpnents and prducts, tissues and bdy fluids; all human and animal cultured cells; all infected animals and animal tissues; all cultures/stcks f bilgical agents, including recmbinant DNA materials; and all bilgical txins. Als includes bimedical waste and physically dangerus (sharp) waste. Decmmissining: The prcess whereby a Principal Investigatr r Labratry Directr and his r her labratry staff decntaminate existing labratry space and make a clinical r research labratry safe prir t vacating the space. Decntaminatin: The prcess whereby the Principal Investigatr r Labratry Directr and his r her labratry staff clean and disinfect labratry surfaces and equipment s they are safe t handle. Rles and Respnsibilities The Principal Investigatr r Labratry Directr is respnsible fr the cmplete decmmissining f the labratry space prir t vacating the labratry. In cases where an abandned lab is identified, the department that the PI r Labratry Supervisr reprted t will be respnsible fr the decmmissining and all csts assciated with the prcess. Envirnmental Health and Safety (EHS) will distribute this plicy and attachments and advise Principal Investigatrs, Labratry Directrs, and labratry persnnel n hw t implement the varius aspects f the plicy. They will als verify that a lab has been apprpriately decmmissined befre a Principal Investigatr r Labratry Directr may leave r mve his r her labratry. Page 179 f 201

180 The Mve Crdinatr fr the labratry is appinted by the Principal Investigatr r Labratry Directr and is respnsible fr crdinating the labratry decmmissining and mve. The Mve Crdinatr is the primary cntact with EHS. Other persnnel (Facilities, mving persnnel, and cntractrs) shuld be aware f this plicy and shuld nt handle labratry materials, equipment, r waste unless instructed t d s by their supervisr and/r EHS. Prcedures: Preparatin Prir planning is key t a successful labratry decmmissining and mve. Preparatin and cmmunicatin with EHS will be a majr factr in minimizing delays, prtecting prperty against damage and lss, and mst imprtantly, reducing the ptential fr persnal injury. Cntact EHS at (617) with any questins r fr assistance. Prcedures: Waste Dispsal All bilgical waste and hazardus waste must be dispsed f accrding t current EHS plicies and prcedures as utlined in the EHS Plicy Manual. All radiactive waste must be dispsed f accrding t Radiatin Prtectin Office plicies and prcedures. Bxes and trash must nt be left in crridrs. Prir arrangements fr regular trash must be made with Custdial r Envirnmental Services. Chemical waste must be labeled with hazardus waste stickers regardless f whether r nt they are labeled frm the manufacturer. Unwanted, unpened, r uncntaminated chemicals shuld be ffered t ther labs that may be able t use them befre the chemicals are cnsidered fr dispsal. Any unknwn chemical must be identified and labeled as hazardus waste. Fr chemical unknwns that cannt be identified by the Principal Investigatr, Labratry Directr r labratry persnnel, the labratry may be assessed a service fee fr hazardus waste analysis prir t dispsal. Darkrm tanks must be drained and the cntents dispsed f as hazardus waste. Empty cmpressed gas tanks must be returned t the distributr prir t the mve. Mercury thermmeters must be dispsed f as hazardus waste, and vacuum pumps must be drained f il and the il dispsed f as hazardus waste. Prcedures: Decntaminatin All labratry bench-tp surfaces must be decntaminated prir t vacating the labratry, and all labratry equipment that is either remaining in the labratry r being mved t a new labratry must be decntaminated if ptentially cntaminated with bilgical, chemical, r radiactive materials. Page 180 f 201

181 Lab equipment requiring decntaminatin includes, but is nt limited t, animal cages, centrifuges, fermenters, fish tanks, incubatrs, water baths, refrigeratrs, and freezers (if nt mving intact). Fume hds must be decntaminated. Cntact the Industrial Hygienist at (617) fr decntaminatin and certificatin advice. Ntify the Industrial Hygienist if there is any current r past practices that may reveal ptential prblems. Certain chemicals such as perchlric acid and mercury may remain n surfaces r equipment r in building systems. Bilgical safety cabinets and glve bxes that have been used with ptentially infectius materials must be decntaminated using parafrmaldehyde gas befre mving. This must be dne by a qualified utside cntractr. If BSCs are either being mved t new labratry areas r being left behind, cntact the Bilgical Safety Officer at (617) t discuss decntaminatin well in advance f the mve. BSCs that are mved must be re-certified after installatin. Cntact B&V Testing at (781) t arrange fr re-certificatin. An apprpriate disinfectant must be utilized in cases where bilgical materials were in use. A disinfectant is deemed apprpriate if it targets the bilgical materials that were in use in the labratry. In mst cases, 70% alchl, bleach slutin (1:10 made fresh), r a phenlic disinfectant shuld be adequate fr disinfectin f lab furniture and equipment ptentially cntaminated with bilgical materials. Call the Bilgical Safety Officer at (617) with questins r cncerns. A BU Equipment Decntaminatin Recrd sticker must be affixed t all equipment that has been decntaminated. This will allw mving persnnel t safely mve the equipment t the new labratry space. Only equipment with this sticker will be mved. Stickers may be btained frm EHS at (617) The Principal Investigatr r Labratry Directr must cmplete the Labratry Decntaminatin Certificatin Frm and submit the frm t EHS (M-470) when decntaminatin and decmmissining activities are cmpleted. This will allw EHS persnnel t review the decmmissining activities, visit the decmmissined labratry, and alert the apprpriate administrative persnnel that the decmmissining has been perfrmed. Upn receipt f the cmpleted frm, EHS will cntact the Principal Investigatr r Labratry Directr t schedule a tur f the labratry t cnfirm the decmmissining activities. Fr mre infrmatin regarding prper disinfectin r decntaminatin prcedures, cntact EHS at (617) r the Radiatin Prtectin Office at (617) Prcedures: Designatin f New Labratry Space The Principal Investigatr r Labratry Directr must infrm EHS f any new labratry space, s that the apprpriate safety signage may be prvided. The Principal Investigatr is respnsible fr ntifying all applicable Bstn University research cmmittees and utside agencies, as necessary, f the mve t new labratry space. Research Page 181 f 201

182 prjects apprved by the IBC must have updated labratry lcatin infrmatin. USDA Veterinary Service r Plant Service permits are labratry site specific, as are CDC Select Agent registratin permits. Cntact the Bilgical Safety Officer at (617) fr assistance. Prcedures: Packing and Mving Labratry Materials Labratry persnnel are respnsible fr cllecting all packaging items needed befre the mve date. Carts, plastic bags, tweling, r ther cushining, absrbent materials, sealable plastic r plastic-lined bxes, labels (e.g. Fragile, Universal Bihazard, ID, Lcatin, Cautin, Radiactive Material), sturdy tape, and spill kits shuld be readily accessible. Each cntainer r piece f equipment must be labeled. Labels must identify the agent, hazard, and necessary precautins. The Principal Investigatr r Labratry Directr is respnsible fr establishing safety and emergency prcedures fr all phases f the mve. Ptential emergencies include material spills, fires, slips and falls, and cuts. Prtective clthing and spill absrbent materials must be available during packing, mving, and unpacking. Prcedures: Packing and Mving Labratry Chemicals In rder t minimize the amunt f chemicals that need t be packed and mved, new chemicals shuld be rdered nly as necessary and in small quantities. Labratry persnnel shuld plan in advance t minimize the inventry f liquid vlume and weight f materials being mved. In additin, reductin f active materials shuld be planned the week prir t the mve. Labratry chemicals must be packed and mved by an utside cntractr apprved by EHS. Prir t the packing and mving, labratry persnnel are respnsible fr labeling each chemical cntainer with the chemical identity. Cmpressed gas tanks that are t be mved must have regulatrs remved and caps secured prir t mving. If pssible, have ld tanks cllected prir t a mve and arrange fr future tanks t be delivered t the new lcatin. Thermmeters must be remved frm refrigeratrs, water baths, and incubatrs prir t equipment mving. Vacuum pump il must be drained frm pumps prir t equipment mving. Prcedures: Packing and Mving Bilgical Materials Bilgical materials must be apprpriately packed and mved by the labratry persnnel. Regulated materials and bilgical materials include all human, plant, and animal pathgens; all human bld, bld cmpnents and prducts, tissues and bdy fluids; all human and animal cultured cells; all animal carcasses and unfixed animal tissues; all cultures/stcks f bilgical agents including recmbinant DNA materials; and all bilgical txins. Prper packaging cnsists f a primary sealed cntainer placed within a secndary sealed, unbreakable cntainer, with enugh absrbent material in between t cntain and absrb any spill. Sme examples f prper packaging include petri dishes in a plastic sleeve within a plastic-lined Page 182 f 201

183 bx using paper twel spacers; stabs in a sealed Tupperware cntainer with paper twels t cushin vials; sealed tubes in a rack placed int plastic sealable cntainer with enugh paper twels t absrb any spilled cntents; tissue culture dishes placed int a plastic-lined dishpan r a sealable cardbard bx with an absrbent. Freezers can be mved intact, prvided all cntents are in sealed, unbreakable cntainers and the freezer remains clsed. Because shifting f cntents may ccur, enclse lse items in bxes, r fix in sme ther way t avid breakage and spills when the freezer is repened. Other equipment, such as fermenters, refrigeratrs, incubatrs, and bilgical safety cabinets must be empty and decntaminated prir t the mve. Labeling: Once packaged, all bilgical materials must be prperly labeled. Labels must include the name, Principal Investigatr (PI), new lcatin, ID f agent, bisafety level, telephne number fr assistance in the event f any breakage, and a FRAGILE ntice if applicable. Als the universal bihazard label shuld be used whenever packaging a BSL-2 r higher agent. Questins cncerning the bisafety level f bilgical materials r requests fr bihazard labels shuld be directed t the Bisafety Safety Officer at (617) Prcedures: Labratry Furniture and Equipment Furniture: The Mve Crdinatr must be infrmed if there is any furniture f particular cncern (fragile, valuable, requires dismantling) nt already mentined. Different mving cmpanies may have different requirements that shuld be ascertained in advance f the mve. Special Requirements: The Mve Crdinatr must be infrmed in advance f any equipment under service cntract, as well as equipment nt under cntract but requiring servicing and/r special handling. Alarms: Labratry persnnel must discnnect alarms n freezers (if mving intact) and any ther sensr alarms n r befre the day f the mve. Keys and Cmbinatins: Labratry persnnel must keep keys and cmbinatins t lcks readily accessible. Page 183 f 201

184 Appendix T Labratry Dr Signage Bstn University Dr Sign Sample: This is cmpleted in accrdance with the NFPA 704 Hazard Identificatin ratings system (the NFPA "hazard diamnd") fr health, flammability, and instability Page 184 f 201