biosafety booklet

8/4/2019 Biosafety Booklet

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A Guide to Biosafety & Biological Safety Cabinets


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F wThis booklet was developed as a guide to biological sa ety cabinets and provides basicknowledge o biosa ety and biological sa ety cabinets. The in ormation presented isunbiased and generic in nature compiled with help rom experienced microbiologists,engineers and sa ety enclosure users.

I. BIohazards and BIosaFety LeveLsThe word biohazard is a contraction o the words biological and hazard, and defned as:an in ectious agent, or part thereo , presenting a real or potential risk to the well-beingo man, animals and / or plants, directly through in ection or indirectly through disruptiono the environment.

Biosa ety Levels 1 through 4 were established by the Centers or Disease Control (CDC)

and the National Institutes o Health (NIH) and are combinations o laboratory practicesand techniques, sa ety equipment and acilities. All o these levels are appropriate or thebiohazard posed by the agents used and or the laboratory activity.

Bi L l 1Practices, sa ety equipment and acilities appropriate or work with defned and characterizedstrains o viable micro organisms not known to cause disease in healthy adult humans. Thelaboratory is not necessarily separated rom the general tra fc patterns in the building.Work is generally conducted on open bench tops using standard microbiological practices.Special containment equipment or acility design is neither required nor generally used.Laboratory personnel have specifc training in the procedures conducted in the laboratoryand are supervised by a scientist with general training in microbiology or a related science.A biological sa ety cabinet is generally not required or work involving these agents.

Bi L l 2Practices, sa ety equipment and acilities appropriate or work done with a broad spectrumo indigenous moderate-risk agents present in the community and associated with human

disease in varying severity. It di ers rom biosa ety level 1 in that:a) laboratory personnel have specifc training in handling pathogenic agents and are directed

by competent scientists;b) access to the laboratory is limited when work is being conducted;c) extreme precautions are taken with contaminated sharp items; andd) certain procedures in which in ectious aerosols or splashes may be created are conducted

in biological sa ety cabinets or other physical containment equipment. A Class I or ClassII biological sa ety cabinet is highly recommended or work involving these agents.

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A Guide to Biosa ety & Biological Sa ety Cabinets Bi Bi L l


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Bi L l 3Practices, sa ety equipment and acilities appropriate or work done with indigenous orexotic agents with a potential or respiratory transmission which may cause serious andpotentially lethal in ection. More emphasis is placed on primary and secondary barriersto protect personnel in the contagious area, the community, and the environment rom

exposure to potentially in ectious aerosols. A Class I or Class II biological sa ety cabinet isrequired or work involving these agents.

Bi L l 4Practices, sa ety equipment and acilities appropriate or work done with dangerous andexotic agents which pose a high risk o li e threatening disease. May be transmitted viathe aerosol route, and or which there is no available vaccine or therapy. Members othe laboratory sta have specifc and thorough training in handling extremely hazardousin ectious agents and they understand the primary and secondary containment unctionso the standard and special practices, the containment equipment, and the laboratorydesign characteristics. They are supervised by competent scientists who are trained andexperienced in working with these agents. Access to the laboratory is strictly controlled bythe laboratory director. The acility is either in a separate building or in a controlled areawithin a building, which is completely isolated rom all other areas o the building. A specifc

acility operations manual is prepared or adopted. A Class III biological sa ety cabinet isrequired or work involving these agents.

From: Biosafety in Microbiology and Biomedical Laboratories, U.S . Department o Health and Human Services, HHS

publication (CDC) 99-8395, 4th Edition. April 1999. p 11-14.

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II. BIoLogIcaL saFety caBInetsThe terms biological sa ety cabinet and biosa ety cabinet have been widely used to describea variety o containment devices equipped with HEPA flter(s), designed to provide personnelor both personnel and product protection rom biohazardous materials. The terms shouldonly be applied to those devices that meet the requirements o Class I, II, or III specifcations,based on their construction, air ow velocities and patterns, and their exhaust systems.

Major International Standards for Biological Safety Cabinets : American Standard NSF 49 European Standard EN 12469 Australian Standard AS 2252 Japanese Standard JIS K 3800

Table 2. Biological Sa ety Cabinet Classes

Source: Laboratory Biosafety Manual . Second Edition (revised). Interim guidelines. World Health Organization. p32.

* Pressure di erential between chamber and environment.

clI w

v l i(m/ )

r l ai(%)

ex uai (%)

c l Pl umsu u b

ex ual i

BiL l

I US:0.38EN:0.70 0 100 Outside AirInside room /

Hard duct 1, 2 & 3

IIType A1

US:0.38EN:0.40 70 30 Outside Air

Inside room / Thimble duct 1, 2 & 3

IIType A2

US:0.50EN:0.40 70 30 Negative Plenum

Inside room / Thimble duct 1, 2 & 3

IIType B1

US:0.50EN:0.40 30 70 Negative Plenum Hard duct only 1, 2 & 3

IIType B2

US:0.50EN:NA 0 100 Negative Plenum Hard duct only 1, 2 & 3

III Closed: *>0.5WC 0 100 Negative PlenumInside room /

Hard duct 1, 2, 3 & 4

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cl I Bi l i l s c biThe Class I cabinet has the most basic and rudimentary design o all biological sa ety cabinetryavailable today. A stream o inward air moving into the cabinet contains aerosols generatedduring microbiological manipulations. It then passes through a fltration system that trapsall airborne particles and contaminants. Finally, clean, decontaminated air is exhausted rom

the cabinet. The fltration system usually consists o a pre-flter and a HEPA (High E fciencyParticulate Air) flter.

Although the Class I cabinet protects the operator and the environment rom exposure tobiohazards, it does not prevent samples being handled in the cabinet rom coming intocontact with airborne contaminants that may be present in room air. Naturally, there is apossibility o cross-contamination that may a ect experimental consistency. Consequently thescope and application o Class I cabinets is limited and it is largely considered obsolete.

All Class I biological sa ety cabinets are suitable or work with microbiological agents assigned

to biosa ety levels 1, 2 and 3.

cl II Bi l i l s c biLike Class I sa ety cabinets, Class II cabinets have a stream o inward air moving intothe cabinet. This is known as the in ow and it prevents the aerosol generated duringmicrobiological manipulations to escape through the ront opening. However, unlike Class Icabinets, the in ow on Class II cabinets ows through the front inlet grille, near the operator.None of the un ltered in ow air enters the work zone of the cabinet, so the product insidethe work zone is not contaminated by the outside air.

A eature unique to Class II cabinets is a vertical laminar (unidirectional) HEPA-fltered airstream that descends downward from the interior of the cabinet. This continuously ushesthe cabinet interior o airborne contaminants and protects samples being handled withinthe cabinet from contamination and is known as the down ow.

The di erences between the various Class II cabinets available lie primarily with thepercentage o air exhausted to that o air re-circulated rom the common air plenum. Inaddition, di erent Class II cabinets have di erent means o cabinet exhaust. Some cabinetsmay exhaust air directly back to the laboratory, while others may exhaust air through adedicated ductwork system to the external environment.

Despite these di erences, all Class II cabinets, like Class I cabinets, protect both the operatorand environment rom exposure to biohazards. In addition, Class II cabinets also protectproduct samples rom contamination during microbiological manipulations within the cabinetinterior and are all suitable or work with agents assigned to biosa ety levels 1, 2 and 3.

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A Guide to Biosa ety & Biological Sa ety Cabinets Bi l i l s c bi


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cl II t p a (a1/a2) Bi l i l s c biThe Class II Type A biological sa ety cabinet is the most common Class II cabinet. It is alsothe most common sa ety cabinet o all the di erent types available. It has a commonplenum rom which 30% o air is exhausted, and 70% re-circulated to the work area asthe down ow.

Type A cabinets exhaust air directly back to the laboratory, and they may contain positivepressure contaminated plenums. When toxic chemicals must be employed as an adjunct tomicrobiological processes, these cabinets should not be used. Exhaust HEPA fltration onlyremoves airborne aerosols including biohazards, and not chemical umes.

The Class II Type A1 has the positively-pressurized contaminated plenum bordering theambient environment, and there ore is less sa e than the Class II Type A2 that has a negativepressure surrounding the positively pressurized contaminated plenum. In case there is aleakage on the positive plenum, the leaking aerosol will be pulled by the negative pressure

back to the positive plenum, and it will not leak out. Because o the sa ety issue, the TypeA1 design is now considered obsolete. In the A2 cabinet, about 70% o air rom thepositive plenum is recirculated as down ow, and the remaining 30% is discharged to thelab through the exhaust flter.

Fig 1. Class II Type A2 BSC

cl II t p B Bi l i l s c biThe main di erence between Type A and Type B cabinet is: Type B cabinets must be operatedwith an external blower and it exhausts air to the external environment via a dedicatedductwork system. Without the external blower, the cabinets internal blower will blow theair (and microbiological agents) inside the work zone through the ront opening, towardsthe operators ace, creating a dangerous situation. This cabinet is not sel -balancing, in thesense that its own blower can only create down ow, and the cabinet relies on the external

blower to create in ow.

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On all Type B cabinets, environmental protection may be enhanced by installing a scrubbingsystem between the exhaust o the cabinet and the fnal exhaust point outside the buildingto neutralize the chemical umes present in exhaust air.

Although Type B cabinets are commonly used when chemicals are involved in your workprocesses, they theoretically provide an increased level o sa ety as compared to other TypeA cabinets. By exhausting air directly to the external environment, they provide an additional ail-sa e in the event that the regular exhaust HEPA fltration ceases to unction.

cl II t p B1 Bi l i l s c biThe Class II Type B1 biological sa ety cabinet was originally specifed by the American NationalCancer Institute. It has a common plenum rom which 70% o air is exhausted, and 30%re-circulated to the work area as the down ow.

Type B1 cabinets also have a dedicated exhaust eature that eliminates re-circulation whenwork is per ormed towards the back within the interior o the cabinet. Toxic chemicalsemployed as an adjunct to microbiological processes should only be employed i they donot interfere with work when re-circulated in the down ow.

cl II t p B2 Bi l i l s c biIn the Class II Type B2 cabinet all in ow and down ow air is exhausted after HEPA ltrationto the external environment without recirculation within the cabinet. Type B2 cabinets aresuitable or work with toxic chemicals employed as an adjunct to microbiological processesunder all circumstances since no re-circulation occurs. In theory, Type B2 cabinets may be

considered to be the sa est o all Class II biological sa ety cabinets since the total exhaustfeature acts as a fail-safe in the event that the down ow and / or exhaust HEPA ltrationsystems cease to unction normally. However, Class II Type B2 cabinets are, in practice,di fcult to install, balance and maintain.

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Fig 2. Class II Type B2


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cl III Bi l i l s c biThe Class III biological sa ety cabinet provides an absolute level o sa ety, which cannot beattained with Class I and Class II cabinets. All Class III cabinets are usually o welded metalconstruction and are designed to be gas tight. Work is per ormed through glove portsin the ront o the cabinet. During routine operation, negative pressure relative to the

ambient environment is maintained within the cabinet. This provides an additional ail-sa emechanism in case physical containment is compromised.

On all Class III cabinets, a supply o HEPA fltered air provides product protection andprevents cross contamination o samples. Exhaust air is usually HEPA fltered and incinerated.Alternatively, double HEPA fltration with two flters in series may be utilized. Materials aretrans erred into the cabinet using a pass-through unit installed at the side o the workarea. Class III cabinets usually exhaust air back to the laboratory; however, air may also beexhausted via a dedicated ductwork system to the external environment. When a dedicatedductwork system is employed, they are also suitable or work employing toxic chemicals asan adjunct to microbiological processes.

All Class III biological sa ety cabinets are suitable or work with microbiological agentsassigned to biosa ety levels 1, 2, 3 and 4. They are requently specifed or work involvingthe most lethal biological hazards.

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Fig 3. Class III BSC


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hePa & ULPa FilThe HEPA flter is the heart o the biological sa ety cabinet. The HEPA flter is a disposabledry-type flter, constructed o borosilicate microfbers cast into a thin sheet, much like apiece o paper.

HEPA:h igh e fciency Particulate a ir: Modern American-convention HEPA: 99.99%

at 0.3 microns ULPA:Ultra Low Penetration a ir: Modern American-convention ULPA: 99.999%

at 0.12 microns

hePa / ULPa Fil c u i :

Fig 4. Pack is glued into a rame - Frame is gasketed to orm nal assembly

Fig 5. Separator Type Fig 6. Separatorless Type

HEPA/ ULPA ilters are designed to remove a broad range o airborne contaminants,

including: Fine dust Smoke Bacteria (typical size: 500 to 0.3 microns) Soot Pollen Radioactive particles Impurity ion -> can affect Integrated Circuit speed

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III. ProPer seLectIon, InstaLLatIon& saFe Use oF Bscs

a. s l i a Bi l i l s c biA BSC should be selected primarily in accordance with the type o protection needed:product protection; personnel protection against Risk Group 1~ 4 microorganisms; personnelprotection against exposure to radionuclides and volatile toxic chemicals; or a combinationo these. Table 3 shows which BSCs are recommended or each type o protection.

Table 3. Biological Sa ety Cabinet Selection

Source: Laboratory Biosafety Manual . Second Edition (revised). Interim guidelines. World Health Organization. p29.

Volatile or toxic chemicals should not be used in BSCs that recirculate exhaust air to theroom, i.e. Class I that are not ducted to building exhaust systems, or Class II Type A1 orClass II Type A2 cabinets. Class II Type B1 cabinets are acceptable or work with minuteamounts o volatile chemicals and radionuclides. A Class II Type B2 cabinet, also called a

total exhaust cabinet, is necessary when signifcant amounts o radionuclides and volatilechemicals are expected to be used.

t p p i Bsc s l i

Personnel Protection, microorganisms in RiskGroups 1-3 Class I, Class II, Class III

Personnel Protection, microorganisms in RiskGroups 4, glove box laboratory Class III

Personnel Protection, microorganisms in RiskGroups 4, suit laboratory Class I, Class II

Product Protection Class II, Class III only if laminar ow included

Volatile radionuclide / chemical protection, minute amounts Class II Type B1, Class II Type A2 vented to the outside

Volatile radionuclide / chemical protection Class I, Class II Type B2, Class III

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B. U i Bi l i l s c bi I t L b

1. L iThe velocity of air owing through the front opening into a BSC is about 0.45 m/s. At thisvelocity the integrity of the directional air in ow is fragile and can be easily disrupted by air

currents generated by people walking close to the BSC, open windows, air supply registers,and opening and shutting doors.

Ideally, BSCs should be situated in a location remote rom tra fc and potentially disturbingair currents. Whenever possible a 30 cm clearance should be provided behind and on eachside o the cabinet to allow easy access or maintenance. A clearance o 30~35 cm abovethe cabinet may be required to provide or accurate air velocity measurement across theexhaust flter and or exhaust flter changes.

2. op

I BSCs are not used properly, their protective benefts may be greatly diminished. Operatorsneed to be careful to maintain the integrity of the front opening air in ow when movingtheir arms into and out o cabinets. Arms should be moved in and out slowly, perpendicularto the ront opening.

Manipulations o materials within BSCs should be delayed or about 1 min a ter placinghands and arms inside to allow the cabinet to adjust and to air sweep the sur ace othe hands and arms. The number o movements across the ront opening should also beminimized by placing all necessary items into the cabinet be ore beginning manipulations.

BSC

BSC BSC

BSCBSC

Return Air Grille

BSC Biological Safety Cabinet

Barrier Airflow

Air Inlet

Personnel Traffic

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A Guide to Biosa ety & Biological Sa ety Cabinets P p s l i , I ll i & s U Bsc

Fig 7. A Typical Class II Cabinet Installation

Cabinet is quite appropriately located with respect to avoidance1.o excessive air movements rom surrounding areas.Cabinet is too close to the doorway and its air low could be2.infuenced by the air inlet too.The airfow o cabinet could also be infuenced by the air inlet. 3.Cabinet is too close to the doorway.4.Cabinet is suitably located provided that the adjacent return air 5.

grille does not infuence cabinet airfow.

Figure 7 illustrates various possible infuences that can be induced by a rooms design and ventilation system on the cabinets airfow. Pleasenote that Figure 7 does not depict any typical installation. In act it is NOT recommended that so many cabinets be placed in a small room, or in close

proximity to each other.

1

2

3

4

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3. M i l Pl m

The ront intake grill o Class II BSCs must not be blocked with paper, equipment or otheritems. Materials to be placed inside the cabinet should be sur ace-decontaminated with70% alcohol. Work may be per ormed on disin ectant-soaked absorbent towels to capturesplatters and splashes.

All materials should be placed as ar back in the cabinet, towards the rear edge o the worksur ace, as practical without blocking the rear grill. Aerosol-generating equipment (e.g.mixers, centri uges, etc.) should be placed towards the rear o the cabinet. Bulky items, suchas biohazard bags, discard pipette trays and suction collection asks should be placed toone side of the interior of the cabinet. Active work should ow from clean to contaminatedareas across the work sur ace.

The autoclavable biohazard collection bag and pipette collection tray should not be placedoutside the cabinet. The requent in-and-out movement needed to use these containers is

disruptive to the integrity o the cabinets air barrier, and can compromise both personneland product protection.

4. op i M i

Most BSCs are designed to permit operation 24 hr/day, and investigators fnd that continuousoperation helps to control the levels o dust and particulate materials in the laboratory.Class II Type A1 and Class II Type A2 BSCs exhausting to the room or connected by thimbleconnections to dedicated exhaust ducts can be turned o when not in use. Other typessuch as Class II Type B1 and Class II Type B2 BSCs, which have hard-duct installations, musthave air ow through them at all times to help maintain room air balance.

Cabinets should be turned on at least 5 min be ore beginning work and a ter completion owork to allow the cabinet to purge, i.e. to allow time or contaminated air to be removed

rom the cabinet environment.

All repairs made on BSCs should be made by a qualifed technician. Any mal unction in theoperation o the BSC should be reported and repaired be ore the BSC is used again.

5. Ul i l Li

Ultraviolet lights are not required in BSCs. I they are used, they must be cleaned weekly toremove any dust and dirt that may block the germicidal e ectiveness o the light. Ultravioletlight intensity should be checked when the cabinet is recertifed to ensure that light emissionis appropriate. Ultraviolet lights must be turned o while the room is occupied, to protecteyes and skin rom inadvertent exposure.

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6. op Fl mOpen ames should be avoided in the near microbe-free environment created inside the BSC.They disrupt the air ow patterns and can be dangerous when volatile, ammable substancesare also used. To sterilize bacteriological loops, micro-burners or electric urnaces areavailable and are preferable to open ames.

7. spill

A copy o the laboratorys protocol or handling spills should be posted, read and understoodby everyone who uses the laboratory. When a spill o biohazardous material occurs within aBSC, cleanup should begin immediately, while the cabinet continues to operate. An e ectivedisin ectant should be used and applied in a manner that minimizes the generation oaerosols. All materials that come into contact with the spilled agent should be disin ectedand / or autoclaved.

8. a u l c if i

The unctional operation and integrity o each BSC should be certifed to national orinternational per ormance standards at the time o installation and regularly therea ter byqualifed technicians, according to the manu acturers instructions.

Evaluation o the e ectiveness o cabinet containment should include tests or cabinetintegrity, HEPA lter leaks, down ow velocity pro le, face velocity, negative pressure/ ventilation rate, air ow smoke pattern, and alarms and interlocks.

Optional tests or electrical leaks, lighting intensity, ultraviolet light intensity, noise leveland vibration may also be conducted. Special training, skills and equipment are required toper orm these tests and it is highly recommended that they are undertaken by a qualifedpro essional.

9. cl i di i i

All items within BSCs, including equipment, should be sur ace-decontaminated and removedrom the cabinet when work is completed, since residual culture media may provide an

opportunity or microbial growth. The interior sur aces o BSCs should be decontaminated

be ore and a ter each use. The work sur aces and interior walls should be wiped with adisin ectant that will kill any microorganisms that might be ound inside the cabinet. At theend o the work day, the fnal sur ace decontamination should include a wipe-down o thework sur ace, the sides, back and interior o the glass. A solution o bleach or 70% alcoholshould be used where e ective or target organisms. A second wiping with sterile water isneeded when a corrosive disin ectant, such as bleach, is used. It is recommended that thecabinet be le t running. I not, it should be run or 5 min in order to purge the atmosphereinside be ore it is switched o .

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10. d mi i

BSCs must be decontaminated be ore flter changes and be ore being moved. The mostcommon decontamination method is by umigation with ormaldehyde gas. In someEuropean country where ormaldehyde is banned, decontamination can be done usingchlorine dioxide or hydrogen peroxide (STERIS and BIOQUELL method). BSC decontamina-

tion should be per ormed by a qualifed pro essional.

11. P l P i equipm

Personal protective clothing should be worn whenever using a BSC. Laboratory coats areacceptable or work being per ormed at Biosa ety Levels 1 and 2. A solid ront, back-closinglaboratory gown provides better protection and should be used at Biosa ety Levels 3 and4 (except or suit laboratories). Gloves should be pulled over the wrists o the gown ratherthan worn inside. Elasticized sleeves can be worn to protect the investigators wrists. Masksand sa ety glasses may be required or some procedures.

12. al m

BSCs can be equipped with one o two kinds o alarm. Sash alarms are ound only oncabinets with sliding sashes. The alarm signifes that the operator has moved the sash toan improper position. Corrective action or this type o alarm is returning the sash to theproper position. Air ow alarms indicate a disruption in the cabinets normal air ow pattern.This represents an immediate danger to the operator or product. When an air ow alarmsounds, work should cease immediately and the laboratory supervisor should be notifed.

Manu acturers instruction manuals should provide urther details. Training in the use oBSCs should cover this aspect.

13. suppl m I m i

Selecting the correct type o BSC, installing it, using it properly and annually certi ying itsoperation are complex processes. It is highly recommended that they proceed under thesupervision o a well-trained and experienced biosa ety pro essional. The pro essional shouldbe highly amiliar with the relevant literature listed in the Re erences section, and shouldhave been trained on all aspects o BSCs. Operators should receive ormal training in the

operation and use o BSCs.

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c. s U Bi l i l s c bi

1. s Up Wear gloves or hand protection Load the cabinet with all items required prior to starting work and sur ace

decontaminate Sur ace decontaminate the work sur ace, side walls and inner back walls Allow the work zone air to purge or a ew minutes be ore commencing work Do not overcrowd the work zone Close the drain valve be ore operation

2. op i Do not obstruct the ront or back air grilles Work as ar in to the cabinet as possible

Minimize arm movement; make slow movements to avoid disrupting cabinet air ow When removing arms rom cabinet be sure to sur ace decontaminate frst, and

move arms out slowly o the cabinet (in direction perpendicular to plane o work zoneopening)

Minimize external air ow disturbances Work rom clean to dirty Biohazard collection bags should be placed inside the cabinet instead o outside Use absorbent pads on the work sur ace where appropriate to minimize splatter and

aerosol generation in case o a spillage

Sur ace decontaminate be ore removing potentially contaminated items rom theinterior Place aerosol-generating instruments as ar in to the interior o the cabinet as possible Clean materials should be at least 150 mm away rom aerosol generating objects to

minimize the chance or cross contamination Hold lids / covers above dishes / sample plates in order to prevent impingement o

downward air Do not use a gas ame whenever possible as it interferes with air ow Put air turbulence generating equipment such as centri uge, blender, or sonicator in

the back 1/3 o the cabinet

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3. Bu BuBurners can cause an out ow of air from inside the cabinet that can endanger the operator.On Class II cabinets, protection against cross contamination may also be compromised. Therehave been incidents where the sash o a recirculating Class II sa ety cabinet was closed whilethe burner was still on, causing heat build up in the cabinet that damaged the flters.

Bunsen Burner Sa ety: An electronic burner which automatically reduces the gas supply to maintain a smaller

ame during periods of non-use An emergency shut o valve should be easily accessible near to the cabinet in case

o an emergency. The burner should be placed as ar into the cabinet working space as possible to

minimize its e ect on cabinet per ormance. A solenoid valve can be interlocked with the gas supply to automatically shut o

the burner

4. s u d wNote: operate the cabinet continuously whenever possible to maximize protection- sur ace decontaminate all items

Seal biohazard bags i used Sur ace decontaminate the cabinet innerside walls, back wall, work sur ace, drain pan

and the inner side o the sliding sash / hinged window Allow work zone air to purge When available: install ront closure or close sash and activate UV lamp (i desired)

5. U M i Pre-flter replacements once every 3 months UV lamp replacement once a year Operational verifcation i appropriate (e.g. alarm checks, basic airfow measurements etc.)

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Iv. BIohazard sPILLs & cLeanUP ProcedUres

a. I i t Bsc: Wait at least fve minutes to allow the BSC to contain aerosols. Wear lab coat, sa ety glasses and gloves during cleanup.

Allow cabinet to run during cleanup. Apply disin ectant and allow a minimum o 20 minutes contact time. Wipe up spillage with disposable disin ectant-soaked paper towel. Wipe the walls, work sur ace and any equipment in the cabinet with a disin ectant-

soaked paper towel. Discard contaminated disposable materials using appropriate biohazardous waste

disposal procedures (e.g. autoclave or BFI). Place contaminated reusable items in biohazard bags, autoclavable pans with lids or

wrap in newspaper be ore autoclaving and cleanup. Expose non-autoclavable materials to disin ectant (20 minute contact time) be ore

removal rom the BSC. Remove protective clothing used during cleanup and place in a biohazard bag or

autoclaving. Run cabinet 10 minutes a ter cleanup be ore resuming work or turning cabinet o .

B. I i t L b, ou i t Bsc: Call the biosa ety o fce i the material is BSL-2 or greater containment. Clear area o all personnel. Wait at least 15 minutes or aerosol to settle be ore

entering spill area. Remove any contaminated clothing and place in biohazard bag to be autoclaved. Put on a disposable gown, sa ety glasses and gloves.

Initiate cleanup with disin ectant as ollows:

1. Place dry paper towel on spill (to absorb liquids); then layer a second set o disin ectantsoaked paper towels over the spill.

2. Encircle the spill with additional disin ectant being care ul to minimize aerosolization

while assuring adequate contact.3. Decontaminate all items within spill the area.4. Allow 20 minutes contact time to ensure germicidal action o disin ectant.5. Wipe equipment with appropriate disin ectant.6. Discard contaminated disposable materials using appropriate biohazardous waste dis-

posal procedures (e.g., autoclave or BFI) .7. Disin ect reusable items.

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A Guide to Biosa ety & Biological Sa ety Cabinets Bi spill & cl up P u


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c. I i t c i u Clear area o all personnel. Wait 30 minutes or aerosol to settle be ore attempting to clean up spill. Wear a lab coat, sa ety glasses and gloves during cleanup. Remove rotors and buckets to nearest biological sa ety cabinet or cleanup.

Thoroughly disin ect inside o centri uge. Discard contaminated disposable materials using appropriate biohazardous wastedisposal procedures (e.g., autoclave or BFI).

d. ou i t L b, I t i Transport labelled biohazardous material in an unbreakable, well-sealed primary

container placed inside o a second unbreakable, lidded container (cooler, plastic panor pail) labelled with the biohazard symbol.

Should a spill occur in a public area, do not attempt to clean it up without appropriatepersonal protective equipment.

Secure the area, keeping all people well clear o the spill. Call pro essional help to assist in cleanup. Stand by during spill response and cleanup activity and provide assistance only as

requested or as necessary.

From: UCSD Biosafety Handbook, UCSD EH&S Biosafety Team 1996. THE UNIVERSITY OF CALIFORNIA, USA. p95-97.

v. reFerences:User & Service Manual. Labculture Class II Type A2 Biohazard Sa ety Cabinet: ESCO Micro Pte Ltd.

Biosafety in Microbiological and Biomedical Laboratories (BMBL) . 4th Edition.O ce o Health and Sa ety (OHS), Centers or Disease Control and Prevention (CDC). US Department o Health.

Laboratory Biosafety Manual . Second Edition (revised). Interim guidelines. World HealthOrganization.

UCSD Biosafety Handbook , UCSD EH&S Research Community or Biosa ety. 1996.

UCSD Biosafety Handbook , UCSD EH&S Biosa ety Team 1996. THE UNIVERSITY OF CALIFORNIA, USA.

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e gl b l o f | Kuala Lumpur, Malaysia | Leiden, The Netherlands | Manama, BahrainMumbai, India | Philadelphia, USA | Salisbury, UK | Shanghai, China | Singapore

Esco Technologies, Inc. 2940 Turnpike Drive, Units 15-16 Hatboro, PA 19040, USAToll-Free USA and Canada 877-479-ESCO Tel 215-441-9661 Fax 215-441-9660us.escoglobal.com [email protected]

Esco Micro Pte. Ltd. 21 Changi South Street 1 Singapore 486 777Tel +65 6542 0833 Fax +65 6542 6920 [email protected]

2 0 0 9 E s c o M i c r o P t e . L t d . S p e c i f c a t i o n s S u b j e c t t o C h a n g e . E S 1 2 8 4 / O W 8 3 5 9 R

_ V 3_ 5 K

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Since 1978, Esco has emerged as a leader in the development ofcontrolled environment, laboratory and cleanroom equipmentsolutions. Products sold in more than 100 countries includebiological safety cabinets, fume hoods, ductless fume hoods,laminar ow clean benches, animal containment workstations,cytotoxic cabinets, hospital pharmacy isolators, and PCR cabinetsand instrumentation. With the most extensive product line in theindustry, Esco has passed more tests, in more languages, for morecerti cations, throughout more countries than any biosafetycabinet manufacturer in the world. Esco remains dedicated todelivering innovative solutions for the clinical, life science, researchand industrial laboratory community. www.escoglobal.com.

Biological Safety Cabinets and Laminar Flow Laboratory Fume Hoods Laboratory OvensLaboratory Incubators PCR Thermal Cyclers Microplate Shaker/Incubators Ultraflow Freezers

biosafety booklet

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