FOR OFFICIAL USE ONLY

Transcription

1 BIOSAFETY MANUAL SOUTH DAKOTA SCHOOL OF MINES & TECHNOLOGY Created: 04/11/09 Updated: 01/30/2017 FOR OFFICIAL USE ONLY

2 Table f Cntents 1.0 SDSM&T Labratry Bisafety Plicy Rles and Respnsibility Principal Investigatr Envirnmental Health and Safety Office General Lab Practices Rutes f Infectin Access Persnal Prtective Equipment (PPE) Eye and Face Prtectin Glves Husekeeping Labratry Bisafety Level Criteria Bisafety Levels 1-4 Definitins Bisafety Level Bisafety Level Bisafety Level Bisafety Level Bisafety Level 1 (BSL 1) BSL 1 Labratry Design and Facilities BSL 1 Standard Micrbilgical Practices BSL 1 Special Practices BSL 1 Special Equipment Bisafety Level 2 (BSL 2) BSL 1 Labratry Design and Facilities BSL 2 Standard Micrbilgical Practices BSL 2 Special Practices BSL 2 Special Equipment Recmmended Wrk Practices Autclaves Autclave Respnsibilities Prcedure Perfrmance Verificatin/Mnitring Mnthly Perfrmance Testing Dcumentatin Sharps

3 5.3 Decntaminatin Decntaminatin Prcesses Bihazardus Waste General Labeling, Packaging and Dispsal Prcedures BSL 1 Specific Waste Prcedures Spill Cleanup Prcedures Spills with NO brken glass/sharps Spills invlving brken glass/sharps Bilgical Spill Kit Cntents Appendix A - Sample Bihazard Ntificatin Sign Appendix B - Sample Sharps Safety Sign Appendix C References The guidelines develped by the Natinal Institutes f Health (NIH), Centers fr Disease Cntrl and Preventin (CDC), Occupatinal Safety and Health Administratin (OSHA) are key cmpnents f this bisafety manual. 3

4 1.0 SDSM&T Labratry Bisafety Plicy The Schl f Mines ffice f Envirnmental Health & Safety (EHS) has develped this manual t serve as a supplement t the campus Lab Safety Prgram, specifically addressing bilgical safety matters. The manual is part f Schl f Mines bisafety prgram, which was established t accmplish the fllwing gals: Prtect persnnel frm expsure t infectius agents Prevent envirnmental cntaminatin Prvide an envirnment fr high quality research while maintaining a safe wrk place Cmply with applicable federal, state and lcal requirements The bisafety manual prvides university-wide safety guidelines, plicies and prcedures fr the use and manipulatin f bihazards. Planning fr and implementatin f bilgical safety must be part f every labratry activity in which bihazard materials are used. In general, the handling and manipulatin f bilgical agents and txins requires the use f varius precautinary measures depending n the material(s) invlved. This manual will prvide assistance in the evaluatin, cntainment and cntrl f bihazards. Hwever, it is imperative that all parties invlved r wrking with these materials seek additinal advice and training when necessary. Currently the Schl f Mines campus cnducts research that requires Bisafety Level 1 and 2 (BSL 1 and BSL 2) cntainment nly. Therefre, nly the descriptins and assignments fr BSL 1 and 2 are detailed herein. 2.0 Rles and Respnsibility 2.1 Principal Investigatr The Principal Investigatr has primary respnsibility fr assuring the safety f the labratry persnnel under his/her directin. This respnsibility includes: Registratin f ptentially hazardus agents with the EHS Office; Assessment f the risks assciated with the agents used and selectin f apprpriate safeguards; Preparatin f a written safety plan; Training and supervisin f staff and students in safe practices; Reprting f accidents and expsures f labratry persnnel t the EHS Office. 2.2 Envirnmental Health and Safety Office The EHS Office is respnsible fr the bilgical safety prgram within the university and fr implementatin f plicies established by the university. The duties include the fllwing: Prviding general surveillance ver activities invlving bihazardus agents, including rutine inspectins f all areas in which bihazardus agents are used; Determining cmpliance with university plicies and safety guidelines; 4

5 Mnitring and reviewing the perfrmance and maintenance f cntainment systems designed fr ccupatinal r envirnmental prtectin; Prviding cnsulting services n aspects f bilgical safety t persnnel at all levels f respnsibility; Prviding guidance and assistance cncerning the packaging and shipping f bihazardus agents leaving the institutin; Maintaining recrds f agents used n campus, their risk classificatin, lcatin and Principal Investigatr; This is accmplished using the SD Mines Bisafety Prtcl frm. Serving as liaisn with NIH, CDC, and ther research rganizatins n matters pertaining t bilgical safety; 3.0 General Lab Practices The fllwing infrmatin applies t all labratries husing bilgical materials. Infrmatin specific t BSL 1 and BSL 2 will fllw. 3.1 Rutes f Infectin An infectin ccurs when disease-causing micrrganisms enter the human bdy in sufficient numbers and by a particular rute and vercme the bdy's defense system. The fllwing rutes f infectin have been reprted fr labratry-acquired infectins: 1. Thrugh the muth Eating, drinking and smking in the labratry Muth pipetting Transfer f micrrganisms t muth by cntaminated fingers r articles 2. Thrugh the skin Accidental inculatin with a hypdermic needle, ther sharp instrument r glass Cuts, scratches 3. Thrugh the eye Splashes f infectius material int the eye Transfer f micrrganisms t eyes by cntaminated fingers 4. Thrugh the lungs Inhalatin f airbrne micrrganisms 3.2 Access When prcedures are in prgress, the lab dr shuld be shut and when n ne is present in the lab the drs shuld be lcked. Anyne requesting access t the labratry, as with any labratry, shuld be questined as 5

6 t their purpse. 3.3 Persnal Prtective Equipment (PPE) Persnal prtective equipment is used t prtect persnnel frm cntact with hazardus materials and infectius agents. Apprpriate clthing may als prtect the materials frm cntaminatin. Persnal prtective devices and safety equipment, as well as training in the prper use f thse devices and equipment, must be prvided t all persnnel under the apprpriate circumstances. The persnnel have the respnsibility f prperly using the equipment Eye and Face Prtectin Safety glasses must be wrn in the lab whenever prcedures are underway invlving a prbability f splash, wrk with lw hazard chemicals, r an impact hazard. Whenever pssible, lab peratins shuld be perfrmed in cntainment devices such as a bilgical safety cabinet r fume hd, r behind a bench-tp shield in rder t minimize the ptential fr skin r mucus membrane cntact with a hazardus splash. If prcedures d nt permit cntainment f the hazard with a cntainment device, then apprpriate PPE must be wrn as utlined: Splash gggles are the nly frm f eye prtectin apprved fr splash hazards. If a chemical (including bleach) r bilgical splash hazard exists, splash gggles must be wrn. Full face prtectin (i.e., face shield) must be used fr prcedures that have anticipated splashes r sprays f infectius r ther hazardus materials t the face r if there is a high ptential fr aersl generatin. Face shields are nt a replacement fr eye prtectin Glves Glves must be selected based n the hazards invlved and the activity t be cnducted. Glves must be wrn when wrking with bihazards, txic substances, hazardus chemicals and ther physically hazardus agents. Temperature resistant glves must be wrn when handling ht material r dry ice. Delicate wrk requiring a high degree f precisin dictates the use f thin walled glves. When latex glves have been chsen, alternatives shuld be made available. Glves shuld be changed as sn as pssible after they have becme cntaminated; when their integrity has been cmprmised r when necessary. Hands shuld be prperly washed with sap and water after remving glves. Dispsable glves must nt be washed r reused. Glves shuld be remved and hands washed when wrk with ptentially infectius materials is cmplete r when leaving the labratry. If yu are transprting ptentially infectius materials (i.e., cultures, waste, etc.) t anther part f the building use the ne glve rule: use ne glved hand fr handling the materials and use the ther unglved hand fr tuching cmmn surfaces such as dr knbs and elevatr buttns. 6

7 3.4 Husekeeping Gd husekeeping in labratries is essential t reduce risks and prtect the integrity f bilgical experiments. Rutine husekeeping must be relied upn t prvide wrk areas free f significant surces f cntaminatin. Husekeeping prcedures shuld be based n the highest degree f risk t which persnnel and experimental integrity is apprpriate. Labratry persnnel are respnsible fr cleaning labratry benches, equipment and areas that require specialized technical knwledge. T facilitate decntaminatin, the labratry shuld be kept neat and free f clutter - surfaces shuld be clean and free f infrequently used chemicals, glassware and equipment. Access t sinks, eyewash statins, emergency shwers and exits, and fire extinguishers must nt be blcked. 4.0 Labratry Bisafety Level Criteria 4.1 Bisafety Levels 1-4 Definitins Bisafety Level 1 Suitable fr wrk invlving well-characterized agents nt knwn t cause disease in healthy adult humans and f minimal ptential hazard t labratry persnnel and the envirnment. Examples: Bacillus subtilis, Naegleria gruberi Infectius canine hepatitis virus, Bisafety Level 2 Suitable fr wrk invlving agents f mderate ptential hazard t persnnel and the envirnment Examples: Measles virus, Salmnellae, Txplasma species, Hepatitis B virus, primary human cells, viral vectrs Bisafety Level 3 Suitable fr wrk with infectius agents which may cause serius r ptentially lethal disease as a result f expsure by the inhalatin rute. Examples: Mycbacterium tuberculsis, St. Luis encephalitis virus, Cxiella burnetii Bisafety Level 4 Suitable fr wrk with dangerus and extic agents that pse a high individual risk f aersl transmitted labratry infectins and life threatening disease. Examples: Ebla Zaire virus, Rift Valley Fever virus 4.2 Bisafety Level 1 (BSL 1) 7

8 Bisafety Level 1 is suitable fr wrk invlving well-characterized agents nt knwn t cnsistently cause disease in healthy adult humans, and f minimal ptential hazard t labratry persnnel and the envirnment. The labratry is nt necessarily separated frm the general traffic patterns in the building. Wrk is generally cnducted n pen bench tps using standard micrbilgical practices. Special cntainment equipment r facility design is neither required nr generally used. Labratry persnnel have specific training in the prcedures cnducted in the labratry and are supervised by a scientist with general training in micrbilgy r a related science BSL 1 Labratry Design and Facilities The facilities required in a bisafety level 1 labratry include the fllwing: Drs Drs are required fr access cntrl. They shuld be kept lcked when n ne is present in the labratry. Sink A sink must be available and supplied fr hand washing (i.e., stcked with sap and paper twels). Easily cleanable Lab must be designed in a way that allws it t be cleaned easily. Carpets and rugs are nt allwed. Spaces between benches, cabinets and equipment must be accessible fr cleaning. Furniture Furniture in the lab must be apprpriate fr the anticipated use. Bench tps must be impervius t water and resistant t heat, rganic slvents, acids, alkalis and ther chemicals. Chairs used in cnjunctin with lab wrk must be cvered with a nn-prus material that can be easily cleaned and disinfected. Windws If the lab has windws that can be pened t the utdrs, they must be fitted with screens BSL 1 Standard Practices Access t the labratry is limited r restricted at the discretin f the Principal Investigatr when experiments are in prgress. Wrk surfaces are decntaminated at least nce a day and after wrk with infectius materials is finished, and after any spill f viable material is cleaned with disinfectants that are effective against the agents f cncern. All cntaminated liquid r slid wastes are decntaminated befre dispsal. Mechanical pipetting devices are used; muth pipetting is prhibited. Plicies fr the safe handling f sharps are instituted. Needles shuld nt be bent, sheared, replaced in the needle sheath r guard, r remved frm the syringe fllwing use. The needle and syringe shuld be prmptly placed in a puncture-resistant cntainer and decntaminated, preferably autclaved. Eating, drinking, smking, and applying csmetics are nt permitted in the wrk area. Fd may be stred in cabinets r refrigeratrs designated and used fr this purpse nly. N preparatin, strage r cnsumptin f fd r drink is permitted in the lab. 8

9 Persns wash their hands: after handling materials invlving rganisms cntaining recmbinant DNA mlecules and animals befre exiting the labratry All prcedures are perfrmed carefully t minimize the creatin f splashes r aersls. A bihazard sign must be psted at the entrance t the labratry whenever infectius agents are present. The sign must include the name f the agent(s) in use and the name and the phne number f the investigatr. Please see Appendix A fr an example f an apprpriate hazard warning sign BSL 1 Special Practices Cntaminated materials that are t be decntaminated at a site away frm the labratry are placed in a durable, hard-walled, leak-prf cntainer that is clsed befre being remved frm the labratry BSL 1 Special Equipment Special cntainment equipment is generally nt required fr manipulatins f agents assigned t BSL 1. Glves shuld be wrn if the skin n the hands is brken r if a rash is present. Prtective eyewear shuld be wrn fr cnduct f prcedures in which splashes f micrrganisms r ther hazardus materials is anticipated. 4.3 Bisafety Level 2 (BSL 2) Bisafety Level 2 builds upn BSL 1. BSL 2 is suitable fr wrk invlving agents that pse mderate hazards t persnnel and the envirnment. It differs frm BSL 1 in that 1) labratry persnnel have specific training in handling pathgenic agents and are supervised by scientists cmpetent in handling infectius agents and assciated prcedures; 2) access t the labratry is restricted when wrk is being cnducted; and 3) all prcedures in which infectius aersls r splashes may be created are cnducted in BSCs r ther physical cntainment equipment. The fllwing standard and special practices, safety equipment, and facility requirements apply t BSL 2: BSL 2 Labratry Design and Facilities Eye wash statins An eyewash statin must be readily available. Decntaminatin A methd fr decntaminating all labratry wastes shuld be available in the facility (e.g., autclave, chemical disinfectin, incineratin, r ther validated decntaminatin methd). Drs Self-clsing drs are required fr access cntrl. They shuld be kept lcked when n ne is present 9

10 in the labratry. Sink A sink must be available and supplied fr hand washing (i.e., stcked with sap and paper twels). The sink may be manually perated, hands free, r autmatically perated. It shuld be lcated near the exit dr. Easily cleanable The lab must be designed in a way that allws it t be cleaned easily. Carpets and rugs are nt allwed. Spaces between benches, cabinets and equipment must be accessible fr cleaning. Furniture Furniture in the lab must be apprpriate fr the anticipated use. Bench tps must be impervius t water and resistant t heat, rganic slvents, acids, alkalis and ther chemicals. Chairs used in cnjunctin with lab wrk must be cvered with a nn-prus material that can be easily cleaned and disinfected. Bisafety cabinets BSCs must be installed s that fluctuatins f the rm air supply and exhaust d nt interfere with prper peratins. BSCs shuld be lcated away frm drs, windws that can be pened, heavily traveled labratry areas, and ther pssible airflw disruptins. HEPA filtered exhaust air frm a Class II BSC can be safely re circulated back int the labratry envirnment if the cabinet is tested and certified at least annually and perated accrding t manufacturer s recmmendatins. BSCs can als be cnnected t the labratry exhaust system by either a thimble (canpy) cnnectin r a direct (hard) cnnectin. Prvisins t assure prper safety cabinet perfrmance and air system peratin must be verified. Vacuum lines Vacuum lines shuld be prtected with High Efficiency Particulate Air (HEPA) filters, r their equivalent. Filters must be replaced as needed. Liquid disinfectant traps are required. Ventilatin Systems There are n specific requirements n ventilatin systems. 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. Windws If the lab has windws that can be pened t the utdrs, they must be fitted with screens BSL 2 Standard Micrbilgical Practices Access t the labratry is limited r restricted at the discretin f the Principal Investigatr. Wrk surfaces are decntaminated at least nce a day and after wrk with infectius materials is finished, and after any spill f viable material is cleaned with disinfectants that are effective against the agents f cncern. 10

11 All cntaminated liquid r slid wastes are decntaminated befre dispsal. Materials t be remved frm the facility fr decntaminatin must be packed in accrdance with applicable lcal, state, and federal regulatins. Decntaminate all cultures, stcks, and ther ptentially infectius materials befre dispsal using an effective methd. Depending n where the decntaminatin will be perfrmed, the fllwing methds shuld be used prir t transprt: Materials t be decntaminated utside f the immediate labratry must be placed in a durable, hard-walled, leak prf cntainer and secured fr transprt. Mechanical pipetting devices are used; muth pipetting is prhibited. Plicies fr the safe handling f sharps, such as needles, scalpels, pipettes, and brken glassware must be develped and implemented. Whenever practical, labratry supervisrs shuld adpt imprved engineering and wrk practice cntrls that reduce risk f sharps injuries. Precautins, including thse listed belw, must always be taken with sharp items. These include: Careful management f needles and ther sharps are f primary imprtance. Needles must nt be bent, sheared, brken, recapped, remved frm dispsable syringes, r therwise manipulated by hand befre dispsal. Used dispsable needles and syringes 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. Brken glassware must nt be handled directly. Instead, it must be remved using a brush and dustpan, tngs, r frceps. Plasticware shuld be substituted fr glassware whenever pssible. Eating, drinking, smking, handling cntact lenses, applying csmetics, and string fd fr human cnsumptin must nt be permitted in labratry areas. Fd must be stred utside the labratry area in cabinets r refrigeratrs designated and used fr this purpse. Persns shuld wash their hands: befre exiting the labratry All prcedures are perfrmed carefully t minimize the creatin f splashes r aersls. A bihazard sign must be psted at the entrance t the labratry whenever infectius agents are present. The sign must include the name f the agent(s) in use and the name and the phne number f the investigatr. Please see Appendix A fr an example f an apprpriate hazard warning sign. Ptentially infectius agent hazard infrmatin shuld be psted in accrdance with the institutinal plicy. An effective integrated pest management prgram is required. The labratry supervisr must ensure that labratry persnnel receive apprpriate training regarding their duties, the necessary precautins t prevent expsures, and expsure evaluatin prcedures. Persnnel must receive annual updates r additinal training when prcedural r plicy changes ccur. Persnal health status may impact an individual s susceptibility t infectin, ability t receive immunizatins r prphylactic interventins. Therefre, all labratry persnnel and particularly wmen f child bearing age shuld be prvided with infrmatin regarding immune cmpetence and cnditins that may predispse them t infectin. Individuals having these cnditins shuld be encuraged t self identify t the institutin s healthcare prvider fr apprpriate cunseling and guidance. 11

12 4.3.3 BSL 2 Special Practices All persns entering the labratry must be advised f the ptential hazards and meet specific entry/exit requirements depending n the space. Labratry persnnel must be prvided medical surveillance and ffered apprpriate immunizatins fr agents handled r ptentially present in the labratry. When apprpriate, a baseline serum sample shuld be stred. A labratry specific bisafety manual must be prepared and adpted as plicy. The bisafety manual must be available and accessible. The labratry supervisr must ensure that labratry persnnel demnstrate prficiency in standard and special micrbilgical practices befre wrking with BSL 2 agents. Ptentially infectius materials must be placed in a durable, leak prf cntainer during cllectin, handling, prcessing, strage, r transprt within a facility. Labratry equipment shuld be rutinely decntaminated, as well as after spills, splashes, r ther ptential cntaminatin. Spills invlving infectius materials must be cntained, decntaminated, and cleaned up by staff prperly trained and equipped t wrk with infectius material. Equipment must be decntaminated befre repair, maintenance, r remval frm the labratry. Incidents that may result in expsure t infectius materials must be immediately evaluated and treated accrding t prcedures described in the labratry bisafety safety manual. All such incidents must be reprted t the labratry supervisr. Medical evaluatin, surveillance, and treatment shuld be prvided and apprpriate recrds maintained. Animals and plants nt assciated with the wrk being perfrmed must nt be permitted in the labratry. All prcedures invlving the manipulatin f infectius materials that may generate an aersl shuld be cnducted within a BSC r ther physical cntainment device BSL 2 Special Equipment (Primary Barriers and Persnal Prtective Equipment) Prperly maintained BSCs (preferably Class II), ther apprpriate persnal prtective equipment, r ther physical cntainment devices must be used whenever: Prcedures with a ptential fr creating infectius aersls r splashes are cnducted. These may include pipetting, centrifuging, grinding, blending, shaking, mixing, snicating, pening cntainers f infectius materials, inculating animals intranasally, and harvesting infected tissues frm animals r eggs. High cncentratins r large vlumes f infectius agents are used. Such materials may be centrifuged in the pen labratry using sealed rtr heads r centrifuge safety cups. Prtective labratry cats, gwns, smcks, r unifrms designated fr labratry use must be wrn while wrking with hazardus materials. Remve prtective clthing befre leaving fr nn labratry areas (e.g., cafeteria, library, administrative ffices). Dispse f prtective clthing apprpriately, r each department is respnsible t determine an alternative methd fr safely laundering.. It is recmmended that labratry clthing nt be taken hme. Eye and face prtectin (gggles, mask, face shield r ther splatter guard) is used fr anticipated splashes r sprays f infectius r ther hazardus materials when the micrrganisms must be handled utside the BSC r cntainment device. Eye and face prtectin must be dispsed f with 12

13 ther cntaminated labratry waste r decntaminated befre reuse. Persns wh wear cntact lenses in labratries shuld als wear eye prtectin. Glves must be wrn t prtect hands frm expsure t hazardus materials. Glve selectin shuld be based n an apprpriate risk assessment. Alternatives t latex glves shuld be available. Glves must nt be wrn utside the labratry. In additin, BSL 2 labratry wrkers shuld: Change glves when cntaminated, integrity has been cmprmised, r when therwise necessary. Wear tw pairs f glves when apprpriate. Remve glves and wash hands when wrk with hazardus materials has been cmpleted and befre leaving the labratry. D nt wash r reuse dispsable glves. Dispse f used glves with ther cntaminated labratry waste. Hand washing prtcls must be rigrusly fllwed. Eye, face and respiratry prtectin shuld be used in rms cntaining infected animals as determined by the risk assessment. 5.0 Recmmended Wrk Practices 5.1 Autclaves This prgram establishes the fllwing requirements: All ptentially infectius materials and ther bihazardus waste generated thrugh research and teaching activities must be effectively decntaminated prir t dispsal. Decntaminatin f materials must be verified using standard and apprved prcedures Autclave Respnsibilities Respnsible Technicians This persn is identified by the department wh maintains the autclaves fr that space. Prvide autclave-specific training directly and schedule prgram management training, as needed. Develp and maintain standard perating prcedures fr prper autclave use and management. Ensure that all dcumentatin is current and accurate. Mnitr autclave use. Serve as the cntact persn and primary resurce fr answering questins and reslving prblems related t the autclave. Ensure all verificatin testing is perfrmed and maintain assciated testing supplies. Ensure that user lg sheets, cycle charts, safety warnings, peratinal ntices, and cntact infrmatin fr respnsible parties are psted near the autclave Autclave Users Participate in required training. Fllw safe and apprpriate autclave use prcedures. Cmplete autclave use lg. Prmptly reprt prblems r misuse t the Respnsible Technician. When in dubt, ask questins. 13

14 5.1.2 Prcedure Autclaving Precautins Autclaving, r steam sterilizatin, is the mst dependable prcedure fr the destructin f all frms f micrbial life. Prper temperature and expsure time are critical factrs in ensuring the reliability f this methd. These critical factrs are dependent upn steam penetratin t every part f the waste lad. Therefre, the autclave user must be mindful t prevent the entrapment f air. If all the air is nt allwed t escape frm the waste during the cycle, steam will nt replace the air. Saturated steam is emplyed under pressure (at least 15 punds per square inch) t achieve a chamber temperature f at least 121 C (250 F) fr a minimum f 30 minutes r as yur prcedure requires. This time is measured after the temperature f the steam saturated material being sterilized reaches 121 C. The hazards assciated with autclaves include extreme heat and high pressure, as well as large, heavy drs and lading carriage. When perating an autclave the fllwing safety prcedures must be fllwed: 1. Becme familiar with the autclave s wner s manual. Thugh the principle is the same fr each, manufacturer recmmendatins fr use can vary widely. 2. Firmly lck autclave drs and gaskets in place befre yu run the autclave t prevent a sudden release f high pressure steam. Sme autclaves d nt have safety interlcks that prevent the autclave frm running if the dr isn t clsed prperly. If yur autclave des nt have safety interlcks, yu will need t take additinal precautins t ensure that the drs are clsed. 3. If yu have an lder autclave that has little r n heat shielding arund the utside, attach signs warning f Ht Surfaces, Keep Away n r next t the autclave t remind peple f the hazard. D nt stack r stre cmbustible materials (cardbard, plastic, vlatile r flammable liquids, cmpressed gas cylinders) next t an autclave. 4. D nt autclave txic, vlatile r radiactive material. If yu have bihazard waste that cntains any f these materials, please cntact EHS fr guidance. 5. When a cycle is cmplete, wait until yu are sure the pressure gauge registers 0 PSI befre pening the dr f the autclave. 6. Wait at least 30 secnds after pening the dr befre reaching r lking int the autclave. 7. Open the dr slwly, keeping head, face, and hands away frm the pening. 8. Allw cntents t cl befre remving them frm the autclave. 9. Remve slutins frm the autclave slwly and gently; sme slutins can bil ver when mved r when expsed t rm temperature. Thick, heat resistant glves, safety gggles r face-shield and a rubber aprn must be wrn when remving ht liquids frm the autclave. Liquids shuld stand fr ver 1 hur befre being handled withut heat resistant glves. 10. Clean up any spills immediately. 11. Reprt any malfunctins r accidents t yur supervisr immediately. Prcedures fr Schl f Mines autclaves: Strng xidizing material (chemicals) must nt be autclaved with rganic material due t an increased risk f explsin. All bihazardus waste must be placed in range bihazard bags with a heat sensitive "Autclaved" indicatr. Prir t autclaving, a bihazard bag cntaining waste must be kept clsed t prevent airbrne cntaminatin and nuisance drs. Hwever, when autclaving, the bag must be pen t allw the steam t penetrate. Upn remval f the bag frm the autclave, it shuld be clsed and dispsed f in an paque (black) waste bag It is recmmended t add water t each bag befre autclaving Autclave bihazardus materials using the recmmended parameters psted n the autclave 14

15 Cycle Optimizatin Factrs which must be cnsidered fr determining cycle types and times include: Shape/size/vlume f cntainers and materials, Thermcnductive prperties f cntainers and cntents, Density f slid materials and viscsity f liquids, Psitin f lad within the chamber, and Lad cnfiguratin. Fr mst bihazardus waste, the autclave must reach and maintain the fllwing cnditins at the mst difficult lcatin f the lad t heat (e.g., inside f a package, autclave bag): 121 C and 15 psi fr at least 30 minutes. Hwever, lnger lengths f time r greater temperatures r pressures may be needed fr sme materials. Because the entire waste lad must be expsed t the apprpriate cnditins, denser lads will require lnger run times fr effective steam/heat penetratin. Cycle parameters fr such lads must be develped, primarily by adjusting cycle times, s that successful sterilizatin is achieved. Waste decntaminatin lads shuld always be run separately in the autclave frm lads f gds t be sterilized fr use. Fr every waste lad that is decntaminated, regardless f cycle type, prcessing time, lad cmpsitin r lad cnfiguratin, a Class 5 Chemical Indicatr (CI) must be used t determine sterilizatin effectiveness. This measure is necessary t ensure that waste, after autclaving, can be handled and dispsed f safely, and thus prtects the health f research persnnel as well as the general public Lad Cnfiguratin Always cnfigure a waste lad in a manner that avids stacking, crwding, and tuching the sides f the autclave chamber. Lad the waste t allw fr air flw arund waste cntainers. Once a cmmnly-used lad cnfiguratin has been prven t decntaminate waste successfully, it is recmmended that it be dcumented and psted fr reference by ther autclave users t save time, resurces, and utilities Lad Cmpsitin and Preparatin Primary cntainers must nt be filled beynd 75% f hlding capacity and must be made f materials that will remain structurally intact during autclaving. All primary cntainers must allw steam penetratin thrugh lsened, vented penings r permeable materials. Secndary cntainers must be used which are deep enugh t cntain spills (e.g., stainless steel pans). 15

16 SPECIAL CONSIDERATIONS FOR LIQUIDS: Bttles r flasks f liquid waste, as well as liquids t be sterilized fr further use, must ALWAYS be run using a Liquids Cycle that includes a slw exhaust t avid bil-ver. D nt decntaminate slid waste and liquid waste (in bttles r flasks) tgether in the same lad. Minute vlumes f liquid waste (e.g., in Petri dishes, cnical tubes, micrtiter plates, r vials), when bagged, can be mixed with slid waste (e.g., dispsable glves, paper twels) and autclaved successfully using a Slids/Gravity cycle r Pre-vacuum cycle. D nt autclave liquid waste cntaining hazardus chemicals r xidizers Perfrmance Verificatin/Mnitring Each lad prcessed in an autclave must meet the perating parameters set up fr that particular cycle, determined by the use f a Class 5 CI. Any lad which fails t be expsed t the perating parameters must be prcessed again. Perfrmance verificatin per lad must ccur as fllws: An externally placed, Class1 CI (i.e., autclave indicatr tape) must be used with each cntainer, bag, r item prcessed in rder t distinguish it frm unprcessed materials. Fr every lad, the user must verify and dcument perating parameters using an internally placed Class 5 CI. Chemical Indicatrs must be 1) within a cmmercially available challenge test pack, r 2) placed in a central psitin within the lad. Prducts are available which aid in the placement and retrieval f CIs frm within bags f waste. Self-fashined test packs which simulate cmmercial prducts can be tested and used with prir apprval Mnthly Perfrmance Testing Operating parameters (i.e., time, temperature, and pressure) fr each prgrammed cycle (i.e., cycle type, time, and temperature) used fr decntaminatin must be verified mnthly by BI (bilgic indicatr) testing. T perfrm this verificatin, BI vials must be 1) within a cmmercially available challenge test pack, r 2) placed in a central psitin within an actual r mck lad f waste, t ffer sufficient challenge fr steam penetratin Perfrmance Failures Any f the fllwing must be taken as an indicatin that sterility was nt achieved: Printut f prcess parameters shws sterilizatin cnditins nt met, Autclave malfunctins, abrted cycles, r alarms during a cycle, Failure f mnthly Bilgical Indicatr Sterility Test, r Failure f CIs t shw adequate prcessing ccurred. 16

17 Failure Respnses Autclave malfunctin (e.g., printut, alarm, abrted cycle, pwer failure) The Respnsible Technician must be ntified immediately and the autclave taken ut f service fr waste prcessing until the prblem is rectified. The Respnsible Technician will determine if service persnnel are required t restre functin and will arrange fr repairs, if needed. Failure f a Bilgical Indicatr Test The Respnsible Technician must be immediately ntified f any BI test failure and the autclave taken ut f service fr waste prcessing until the cause f the failure is fund and rectified. Psitive test results frm BI can result frm a variety f causes, such as inadequate steam quality, insufficient expsure time r temperature, pr lading practices, r prduct failure r peratr failure. All pssible causes shuld be investigated by the Respnsible Technician. Prper autclave functin must be verified using repeat BI testing by the Respnsible Technician. Once verified, the autclave may be returned t service fr waste prcessing. Failure f Chemical Indicatrs (Class 1 r 5) The Respnsible Technician must be immediately ntified f any CI failure and the autclave taken ut f service fr waste prcessing until the cause f the failure is fund and rectified. CI failures can result frm a variety f causes, such as inadequate steam quality, insufficient expsure time r temperature, pr lading practices, r prduct failure r peratr failure. All pssible causes shuld be investigated by the Respnsible Technician. Prper autclave functin must be verified using repeat CI and/r BI testing by the Respnsible Technician. Once verified, the autclave may be returned t service fr waste prcessing Dcumentatin Autclave Use Lg Must be maintained separately fr each autclave. Must be lcated in clse prximity t the autclave. All runs must be dcumented. Service Lg Must be maintained separately fr each autclave. Respnsible technician must dcument maintenance, repair, and calibratin services in this lg. Bilgical Indicatr Test Results Lg Must be maintained separately fr each autclave. New autclave and mnthly test results must be recrded in this lg. 17

18 5.2 Sharps Generally, the use f sharps shuld be restricted t prcedures fr which there is n alternative. Plastic alternatives shuld be substituted fr glassware whenever pssible t prevent the unnecessary ptential fr sharps related expsure incidents. If it has been determined that the use f sharps is unavidable, the fllwing practices shuld be adhered t: All persnnel shuld be trained in safe sharps handling prcedures. Use dispsable sharps devices (i.e., scalpels, prbes, needles) if at all pssible. Prcedures shuld be rganized in a manner that limits persnnel expsure t the sharp device. Fr example: D nt expse/unsheathe sharp devices until the prcedure actually requires the use f these items. D nt leave expsed sharp items unattended. If feasible, place an apprved sharps cntainer within arm's reach f the pint f use fr the sharp item t allw fr immediate dispsal (fr reusable sharps, use a hard-walled cntainer that enclses the sharp end f the device). D nt bend r break sharps. D nt recap sharps if pssible. If recapping is required, use a ne-handed scp technique. Nte: The need fr recapping can be eliminated thrugh the use f safer sharps devices. D nt handle sharps with tw hands. Dispse f waste sharps in a prperly labeled apprved sharps cntainer. Permanently clse and dispse f sharps cntainers when they are ¾ full r within 90 days f the date f first use, whichever cmes first. D NOT verfill r shake cntainers because these actins can result in accidental sharps expsure. Reusable sharps shuld be placed in a hard walled cntainer fr strage until prcessing fr reuse. Brken glassware shuld be handled with a mechanical device, such as tngs, frceps, r a brm and dustpan rather than directly by hand. 5.3 Decntaminatin Decntaminatin is defined as the reductin f micrrganisms t an acceptable level. Methds applied t reach this gal can vary and mst ften include disinfectin r sterilizatin. Generally speaking, disinfectin is used t make the acceptable level f micrrganisms present is defined as being belw the level necessary t cause disease. This means that viable micrrganisms are still present. In cntrast, sterilizatin is defined as the cmplete killing f all rganisms present. Depending n the circumstances and tasks, decntaminatin f a surface (e.g., lab bench) is accmplished with a disinfectant, while decntaminatin f bimedical waste is dne by sterilizatin in an autclave. Many different terms are used fr disinfectin and sterilizatin. The fllwing are cmmnly used in bisafety: Antimicrbial - An agent that kills micrrganisms r suppresses their grwth and multiplicatin. Antiseptic - A substance that inhibits the grwth and develpment f micrrganisms withut necessarily killing them. Antiseptics are usually applied t bdy surfaces. Bicide - A general term fr any agent that kills rganisms. Chemical germicide - A chemical r a mixture f chemicals used t kill micrrganisms. 18

19 Disinfectant - A chemical r mixture f chemicals used t kill micrrganisms, but nt necessarily spres. Disinfectants are usually applied t inanimate surfaces r bjects. Micrbicide - A chemical r mixture f chemicals that kills micrrganisms. The term is ften used in place f "bicide", "chemical germicide" r "antimicrbial." Sprcide - A chemical r mixture f chemicals used t kill micrrganisms and spres. When chsing a methd f decntaminatin, it is imprtant t cnsider the fllwing aspects: Type f bihazardus agents, cncentratin and ptential fr expsure; Physical and chemical hazards t prducts, materials, envirnment and persnnel Decntaminatin Prcesses Physical and chemical means f decntaminatin fall int fur main categries: Heat In rder t kill micrbial agents, heat can be applied in dry r wet frm. The advantage f wet heat is a better heat transfer t and int the cell resulting in shrter expsure time and lwer temperature. Steam sterilizatin uses pressurized steam at C ( F) fr 30 r 40 minutes. This type f heat kills all micrbial cells including spres, which are nrmally heat resistant. In rder t accmplish the same effect with dry heat in an ven, the temperature needs t be increased t C ( F) fr perids f 2 t 4 hurs. Decntaminatin f Bihazardus Waste by Autclaving Autclaving is accepted as a safe and effective prcedure fr sterilizatin when perfrmed in accrdance with the plicies described in 5.1 Autclaves. T ensure that any bihazardus waste created by the Schl f Mines cmmunity is prperly decntaminated, each autclave will be tested n a mnthly basis as described in Mnthly Perfrmance Testing. Liquid chemicals The apprpriate liquid disinfectant shuld be chsen after carefully assessing the bihazardus agent and the type f material t be decntaminated. Liquid disinfectants are preferably used fr slid surfaces and equipment. They vary greatly in their efficiency, depending n the chemical cnstituents and the agents invlved. Variables t remember when disinfecting: Nature f the surface being disinfected - Prus r smth; the mre prus and rugh the surface, the lnger a disinfectant will need t remain n it t be effective. Number f micrrganisms present - Higher cncentratins require a lnger applicatin time and/r higher cncentratin f disinfectant. Presence f rganic material - The prteins in rganic materials such as bld, bdily fluids, and tissue can prevent r slw the activity f certain disinfectants. Duratin f expsure and temperature - Increased expsure time increases the effectiveness f disinfectants. Lw temperatures may slw dwn the activity requiring mre expsure time. EPA regulatin f disinfectants 19

20 The Envirnmental Prtectin Agency (EPA) regulates pesticides, including chemical disinfectants, under the Federal Insecticide, Fungicide, and Rdenticide Act. They are required t be registered with the EPA. It is imprtant t fllw the directins n the manufacturer's label, including thse fr cncentratin and cntact time, when using disinfectants t ensure cmpliance with the EPA requirements. There are many different liquid disinfectants available under a variety f trade names. In general, these can be categrized as halgens, acids r alkalines, heavy metal salts, quaternary ammnium cmpunds, aldehydes, ketnes, alchls, and amines. Unfrtunately, the mst effective disinfectants are ften crrsive and txic. Alchls: Ethyl r isprpyl alchl at a cncentratin f 70% are gd general-use disinfectants. Hwever, they evaprate fast and therefre have limited expsure time. They are less active against nn-lipid viruses and ineffective against bacterial spres. Frmalin: Frmalin is a 37% slutin f frmaldehyde in water. Dilutin f frmalin t 5% results in an effective disinfectant. Frmaldehyde is a suspected human carcingen and creates respiratry prblems at lw levels f cncentratin. Glutaraldehyde: This cmpund, althugh chemically related t frmaldehyde, is mre effective against all types f bacteria, fungi, and viruses. Vaprs f glutaraldehydes are irritating t the eyes, nasal passages and upper respiratry tract. They shuld always be used in accrdance with the instructins n the label and the apprpriate persnal prtective equipment. Phenl and Phenl Derivatives: Phenl based disinfectants cme in varius cncentratins ranging primarily frm 5% t 10%. These derivatives, including phenl, have an dr which can be smewhat unpleasant. Phenl itself is txic and apprpriate persnal prtective equipment is necessary during applicatin. The phenlic disinfectants are used frequently fr disinfectin f cntaminated surfaces (e.g., walls, flrs, bench tps). They effectively kill bacteria including Mycbacterium tuberculsis, fungi and lipid-cntaining viruses. They are nt active against spres r nn-lipid viruses. Quaternary Ammnium Cmpunds (Quats): Quats are catinic detergents with strng surface activity. They are acceptable fr general-use disinfectants and are active against Gram-psitive bacteria and lipid-cntaining viruses. They are less active against Gramnegative bacteria and are nt active against nn-lipid-cntaining viruses. Quats are easily inactivated by rganic materials, aninic detergents r salts f metals fund in water. If Quats are mixed with phenls, they are very effective disinfectants as well as cleaners. Quats are relatively nntxic and can be used fr decntaminatin f fd equipment and fr general cleaning. Halgens (Chlrine and Idine): Chlrine-cntaining slutins have brad spectrum activity. Sdium hypchlrite is the mst cmmn base fr chlrine disinfectants. Cmmn husehld bleach (5% available chlrine) can be diluted 1/10 t 1/100 with water t yield a satisfactry disinfectant slutin. Chlrine cntaining disinfectants are inactivated by excess rganic materials. They are als strng xidizers and very crrsive. Always use apprpriate persnal prtective equipment when using these cmpunds. At high cncentratins and extended cntact time, 20

21 hypchlrite slutins are cnsidered cld sterilants since they inactivate bacterial spres. Idine has similar prperties t chlrine. Idphrs (rganically bund idine) are recmmended disinfectants. They are mst ften used as antiseptics and in surgical saps and are relatively nntxic t humans. Vaprs and Gases A variety f vaprs and gases pssess germicidal prperties. The mst cmmnly used are frmaldehyde and ethylene xide. Applied in clsed systems under cntrlled cnditins (e.g., humidity) these gases achieve sterility. Frmaldehyde gas is primarily used in the decntaminatin f spaces r bilgical cntainment equipment like bilgical safety cabinets. Frmaldehyde is a txic substance and a suspected human carcingen. Cnsiderable cautin must be exercised in handling, string, and using frmaldehyde. Ethylene xide is used in gas sterilizers under cntrlled cnditins. Ethylene xide is als a human carcingen and mnitring is necessary during its use. Incineratin Incineratin is useful fr dispsing f animal carcasses as well as anatmical and ther labratry waste, with r withut prir decntaminatin. 6.0 Bihazardus Waste At SDSM&T, the term bihazardus waste is used t describe different types f waste that might include infectius agents. Currently, the fllwing categries are all cnsidered t be bihazardus waste, as defined by Suth Dakta Administrative Rule 74:35. Medical waste Medical waste means any slid waste which is generated in the diagnsis, treatment, r immunizatin f human beings r animals, in research, r in the prductin r testing f bilgicals. It des nt include any hazardus waste, radiactive waste, r husehld waste. Regulated Medical Waste Regulated medical waste is slid waste generated in the diagnsis, treatment, r immunizatin f human beings r animals, in research pertaining t diseases f humans r animals, r in the prductin r testing f bilgicals, as listed in this sectin: 1) Cultures and stcks: cultures and stcks f infectius agents and assciated bilgicals, including the fllwing: a) Cultures frm medical and pathlgical labratries; 21

22 b) Cultures and stcks f infectius agents frm research and industrial labratries; c) Wastes frm the prductin f bilgicals; d) Discarded live and attenuated vaccines; and e) Culture dishes and devices used t transfer, inculate, and mix cultures; 2) Pathlgical waste: human pathlgical waste, including: a) Tissues, rgans, and bdy parts and bdy fluids that are remved during surgery, autpsy, r ther medical prcedures except thse extracted teeth that are returned t the patient; and b) Specimens f bdy fluids and their cntainers; 3) Human bld and bld prducts, as fllws: a) Liquid waste human bld; b) Prducts f bld; c) Items saturated r dripping with human bld; d) Items that were saturated r dripping with human bld that are nw caked with dried human bld; e) Serum, plasma, and ther bld cmpnents, and their cntainers, which were used r intended fr use in either patient care, testing, and labratry analysis r the develpment f pharmaceuticals; and f) Intravenus bld and bld prduct bags; 4) Sharps: sharps that have been used in animal r human patient care r treatment r in medical, research, r industrial labratries, including the fllwing: a) Hypdermic needles; b) Syringes with the attached needle r cntaining bdy fluids; c) Pasteur pipettes; d) Scalpel blades; e) Bld vials; f) Needles with attached tubing; g) Culture dishes, regardless f the presence f infectius agents; and h) Other types f brken r unbrken glassware that were in cntact with infectius agents, such as used slides and cver slips; 5) Animal waste: cntaminated animal carcasses, bdy parts, and bedding f animals that were knwn t have been expsed t infectius agents during research (including research in veterinary hspitals), prductin f bilgicals, r testing f pharmaceuticals; 6) Islatin waste: bilgical waste and discarded materials cntaminated with bld, excretin, exudates, r secretins frm humans wh are islated t prtect thers frm certain highly cmmunicable diseases as identified by the health care facility r islated frm animals knwn t be infected with highly cmmunicable diseases; 7) Unused sharps: the fllwing unused, discarded sharps: 22

23 a) Hypdermic needles; b) Suture needles; c) Syringes with the attached needle; and d) Scalpel blades. 6.1 General Labeling, Packaging and Dispsal Prcedures The respnsibility fr decntaminatin and prper dispsal f bihazardus waste lies with the prducing facility (e.g., labratry and department). All bihazardus waste needs t be packaged, cntained and lcated in a way that prtects and prevents the waste frm release at any time at the prducing facility prir t ultimate dispsal. If strage is necessary, putrefactin and the release f infectius agents int the air must be prevented. If nt stated therwise, bihazardus waste will be dispsed f in bihazard bags. All waste dispsed f in these bags is t be autclaved in an apprved autclave until the waste is decntaminated. After successful autclaving (decntaminatin), all bihazard bags need t be bagged in paque (black) plastic nn-bihazard bags that are leak prf. These paque bags can be picked up by custdial services. Bihazardus waste that has been successfully decntaminated by autclaving is n lnger cnsidered hazardus. Since autclaves are an integral part f Schl f Mines bihazardus waste treatment prcedure, prper peratin and maintenance as described in 5.1 Autclaves is very imprtant. All users f autclaves need t be trained in the prper perating prcedures by either thrugh the labratry supervisr r the technician assigned t that autclave. Maintenance and repair f autclaves used fr the decntaminatin f bihazardus waste are the respnsibility f the individual departments. If the department chses t nt use autclaves fr their bihazardus waste treatment, alternative prcedures (e.g., utside bimedical waste hauler) need t be established BSL 1-Specific Waste Prcedures Cultures, Stcks and Related Materials Cultures and stcks f infectius agents and assciated bilgicals, shall be placed in bihazard bags and decntaminated by autclaving. Duble r triple bagging may be required t avid rupture r puncture f the bags. Bulk Liquid Waste, Bld and Bld Prducts All liquid waste frm humans r animals such as bld, bld prducts and certain bdy fluids, nt knwn t cntain infectius agents, can be dispsed f directly by flushing dwn a sanitary sewer. Hwever, due t cagulatin, flushing f large quantities f bld is impractical. Cntact the EHS Office fr additinal infrmatin n dispsal f large vlumes f bld. All ther liquid bihazardus waste needs t be autclaved prir t dispsal r treated with a disinfectant. Sharps All sharps must be placed in a rigid, puncture resistant, clsable and leak prf cntainer, which is labeled with 23