General Biosafety
description
Transcript of General Biosafety
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General BiosafetyGeneral BiosafetyTassanee EamkamonBiosafety ProfessionalArmed Forces Research Institute of Medical [email protected]
23 March 2010
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Objectives
• Principle of Biosafety• Overview of Biosafety Levels (BSL) and Risk
Groups (RG)• Biosafty Level 2 Requirement
2
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• A fundamental objective of any biosafety program is the containment of potentially harmful biological agents.
• The purpose of containment is to reduce or eliminate exposure of follows to potentially hazardous agents.
- Laboratory workers- Other persons, - Outside environment
Principle of Biosafety
3www.clker.com/clipart-2970.html
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• The term "containment" is used in describing:
- Safe methods (Practices)
- Safety Equipment (Primary Barriers and Personal Protective Equipment)
- Facilities Design and Construction (Secondary Barrier)
Principles of Biosafety
4
www.pandemicfluonline.com/%3Fp%3D1359
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Biosafety Level?CDC-NIH
http://www.cdc.gov/OD/OHS/biosfty/bmbl5/BMBL_5th_Edition.pdf
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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Biosafety Level (BSL)
• Combination• 4 Levels• Each level is specifically appropriate
for:- Operations performed- Risk Group and documented or suspected
routes of transmission of the infectious agents
6
Safe Practice
Facility Design
Safety Equipment
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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Appropriate for:Undergraduate and secondary educational training and teaching laboratories
Laboratories in which work is done with defined and characterized strains of viable microorganisms not known to consistently cause disease in normal, healthy humans
- Bacillus subtilis- Vaccine strains that have undergone multiple in vivo passages
BSL 1BSL 1Safe
Practice
Facility Design
Safety Equipme
nt
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Appropriate for: Clinical, diagnostic, teaching, and other laboratories
Lab work with a broad spectrum of indigenous moderate-risk agents that are present in the community and associated with human disease of varying severity
Lab work with any human-derived blood, body fluids, tissues, or primary human cell lines where the presence of an infectious agent may be unknown
-Hepatitis B virus - P. faciparum-Salmonellae -H1N1
BSL 2BSL 2Safe
Practice
Facility Design
Safety Equipme
nt
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9
Appropriate for:Clinical, diagnostic, teaching, research, or production facilities
Lab work with indigenous or exotic agents:-with a known potential for aerosol transmission-may cause serious and potentially lethal infection
-Mycobacterium tuberculosis-Chikungunya
BSL 3BSL 3Safe
Practice
Facility Design
Safety Equipme
nt
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10
Appropriate for: Lab work with dangerous and exotic agents which:-- pose a high individual risk of life-threatening disease-- may be transmitted via the aerosol route-- there is no available vaccine or therapy
-Ebola virus- Hendra and Nipah virus
BSL 4BSL 4Safe
Practice
Facility Design
Safety Equipme
nt
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Risk Group?
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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Risk Group (RG)Risk classification based on:
PathogenicityModes of transmissionHost range
Availability of effective preventive measures
Availability of effective treatment
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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Risk Group (RG)
1. WHO (2004)2. NIH Recombinant DNA Guidelines (USA, 2002)3. Australian/New Zealand Standard (2002)4. Canadian Laboratory Safety Guidelines (2004)5. European Economic Community (2000)
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RISK GROUPCLASSIFICATION
NIH GUIDELINES FOR RESEARCHINVOLVING RECOMBINANT DNAMOLECULES 2002
WORLD HEALTH ORGANIZATIONLABORATORY BIOSAFETY MANUAL3RD EDITION 2004
Risk Group 1 Agents that are not associated with disease in healthy adult humans.
Bacillus subtilis or Bacillus licheniformis
A microorganism that is unlikely to cause human or animal disease.(No or low individual and community risk)
Risk Group 2 Agents that are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available.
Campylobacter coli, Salmonella, P. falciparum, Hepatitis viruses
A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment. Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread of infection is limited.(Moderate individual risk; low community risk)
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
www.travelpod.com/travel-blog-en...pod.html www.mrjusino.com/science.htm
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RISK GROUPCLASSIFICATION
NIH GUIDELINES FOR RESEARCHINVOLVING RECOMBINANT DNAMOLECULES 2002
WORLD HEALTH ORGANIZATIONLABORATORY BIOSAFETY MANUAL3RD EDITION 2004
Risk Group 3 Agents that are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available
M. tuberculosis, R. tsutsugamushi, Japanese encephalitis virus, HIV
A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another. Effective treatment and preventive measures are available. (High individual risk; low community risk)
Risk Group 4 Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available
Ebola virus, Tick-borne encephalitis virus complex , Herpesvirus simiae (Herpes B or Monkey B virus)
A pathogen that usually causes serious human or animal disease and that can be readily transmitted from one individual to another, directly or indirectly. Effective treatment and preventive measures are not usually available.(High individual and community risk) available.
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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http://www.absa.org/riskgroups/index.html
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Biosafety Level &
Risk Group
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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Overview of Biosafety Levels (BSL) and Risk Groups (RG)WHO 2004: Laboratory Biosafety Manual
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BSL AGENTS PRACTICES PRIMARY BARRIERS AND SAFETY EQUIPMENT
FACILITIES (SECONDARY BARRIERS)
1 Not known to consistently cause diseases in healthy adults
Standard Microbiological Practices
None required Laboratory bench and sink required
2 • Agents associated with human disease• Routes of transmission include percutaneous injury, ingestion, mucous membrane exposure
BSL-1 practice plus:• Limited access• Biohazard warning signs• “Sharps” precautions• Biosafety manual defining any needed waste decontamination or medical surveillance policies
Primary barriers:• Class I or II BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materialsPPEs:• Laboratory coats; gloves;face protection as needed
BSL-1 plus:• Autoclave available
Summary of Recommended Biosafety Levels for Infectious Agents
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
CDC-NIH 2007: Biosafety in Microbiological and Biomedical Laboratories (BMBL)
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BSL AGENTS PRACTICES PRIMARY BARRIERS AND SAFETY EQUIPMENT
FACILITIES (SECONDARY BARRIERS)
3 • Indigenous or exotic agents with potential for aerosol transmission• Disease may have serious or lethal consequences
BSL-2 practice plus:• Controlled access• Decontamination of all waste• Decontamination of laboratory clothing before laundering• Baseline serum
Primary barriers:• Class I or II BSCs or other physical containment devices used for all open manipulation of agents
PPEs:• Protective laboratory clothing; gloves; respiratory protection as needed
BSL-2 plus:• Physical separation from access corridors• Self-closing, double-door access• Exhaust air not recirculated• Negative airflow into laboratory
Summary of Recommended Biosafety Levels for Infectious Agents
Overview of Biosafety Levels (BSL) and Risk Groups (RG)CDC-NIH 2007: BMBL
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BSL AGENTS PRACTICES PRIMARY BARRIERS AND SAFETY EQUIPMENT
FACILITIES (SECONDARY BARRIERS)
4 • Dangerous/exotic agents which pose high risk of life threatening disease• Aerosol-transmitted laboratory infections have occurred; or related agents with unknown risk of transmission
BSL-3 practices plus:• Clothing change before entering• Shower on exit• All material decontaminated on exit from facility
Primary barriers:• All procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive pressure personnel suit
BSL-3 plus:• Separate building or isolated zone• Dedicated supply and exhaust, vacuum, and decontamination systems• Other requirements outlined in the text
Summary of Recommended Biosafety Levels for Infectious AgentsBiosafty Level 2 Requirement
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Biological risk assessments
• Provide a guide for the selection of appropriate biosafety levels and microbiological practices, safety equipment, and facility safeguards that can prevent LAIs
• Responsibility: Directors and principal investigators of, Institutional biosafety committees (IBC), Animal care and use committees, Biological safety professionals, and Laboratory animal veterinarians
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
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Risk Criteria for Establishing Ascending Levels of Containment
• Infectivity• Severity of disease• Transmissibility• Nature of the work being
conducted• Origin of the agent:
Indigenous or Exotic
Overview of Biosafety Levels (BSL) and Risk Groups (RG)
www.topnews.in/scientists-find-w...-2143806 www.rose-bowl-parade.com/dangers.html
caribbean.scielo.org/scielo.php%...26nrm%3D
www.celprogen.com/store/index.ph...46_59_61
Safe Practice
Facility Design
Safety Equipme
nt
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Reduce/eliminate risk of infection
BSL ?
Agent
Virulence
ExposureExposure
Susceptibility
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Biosafty Level 2
Standard Microbiological PracticesSpecial PracticesSafety EquipmentLaboratory Facilities
Safe Practice
Facility Design
Safety Equipment
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Biosafety Level 2: Standard Microbiological Practices
• Control access to the laboratory.
• Wash their hands after working and before leaving the laboratory.
• Safely handle of sharps.
• Perform all procedures to minimize the creation of splashes and/or aerosols.
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Biosafety Level 2: Standard Microbiological Practices
• Decontaminate work surfaces after completion of work and after any spill or splash.
• Decontaminate all cultures, stocks, and other potentially infectious materials before disposal.
• Implement an effective integrated pest management program.
skitterbot.com/blog/%3Ftag%3Droboticsbhrcivicgroup.blogspot.com/
2009/...hem.html
http://i.ehow.com/images/GlobalPhoto/Articles/4492263/clorox2-main_Full.jpg
Disinfectant
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Biosafety Level 2: Standard Microbiological Practices
• Laboratory supervisor must ensure that laboratory personnel receive appropriate training:
Duties Necessary precautions to
prevent exposures Exposure evaluation
procedures.
• Personnel must receive annual updates or additional training.
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Biosafety Level 2: Standard Microbiological Practices
• Personal health status may impact an individual’s susceptibility to infection, ability to receive immunizations or prophylactic interventions.
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Biosafety Level 2: Standard Microbiological Practices
• Post a sign incorporating the universal biohazard symbol at the entrance to the laboratory. Information must include:
Biosafety level Supervisor’s name (or other
responsible personnel) Telephone number, Required procedures for entering
and exiting the laboratory Agent information
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Biosafety Level 2: Standard Microbiological Practices
Do Not• Eat, drink, smoke, handle
contact lenses, apply cosmetics, and store food for human consumption in laboratory areas.
• Store food outside the laboratory area.
• Mouth pipette.
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Biosafety Level 2: . Special Practices
• Advised all persons entering the laboratory of the potential hazards and meet specific entry/exit requirements.
• Prepare and adopt a laboratory-specific biosafety manual as policy. The manual is available and accessible.
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1. Protect employees, the surrounding community and the environment during activities involving potentially hazardous biological agents.
2. Provide minimum technical safety and occupational health requirements and guidelines for developing and control process for operation involving etiologic agents.
3. Provide an environment for high quality research while maintaining a safe work place.
4. Comply with applicable federal, state and local requirements .
Biological Safety Manual Goals
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Biosafety Level 2: . Special Practices
• Laboratory supervisor must ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices before working.
AFRIMS Department of Enteric Disease Training 2005
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Biosafety Level 2: . Special Practices
• Provided Laboratory personnel medical surveillance and offer appropriate immunizations.
• Store a baseline serum sample, when appropriate.
radiologyinthai.blogspot.com/200...ive.html
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Biosafety Level 2: . Special Practices
• Place potentially infectious materials in a durable, leak proof container during collection, handling, processing, storage, or transport within a facility.
• Decontaminate laboratory equipment routinely.
• Decontaminate equipment before repair, maintenance, or removal from the laboratory.
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Biosafety Level 2: . Special Practices
• Contain, decontaminate, and clean up spills by staff properly trained.
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Biosafety Level 2: . Special Practices
• Immediately evaluated and treated incidents.
• Report all such incidents to the laboratory supervisor.
• Provide medical evaluation, surveillance, and treatment and maintain appropriate records.
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Biosafety Level 2: . Special Practices
• Conduct all procedures that may generate an aerosol in a BSC or other physical containment devices.
• Do not permit animals and plants not associated with the work being performed in the laboratory.
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Biosafety Level 2: Safety Equipment (Primary Barriers and Personal Protective Equipment)
• Use properly maintained BSCs (preferably Class II)
• Appropriate PPEs
• Other physical containment.
www.labdepotinc.com
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• Laminar Flow (LF): - Product protection (no personnel protection) - Not for biohazard agents or chemical fumes
• Fume Hood: - Removes toxic chemical (ducting
sys./ductless) - No HEPA filter -> not for biohazard agents
• Biohazard Safety Cabinet (BSC) - Product and personnel protection (not Class
1)
web.princeton.edu/.../labsafetyman ual/bsc1 .jpg
ateam.lbl.gov/.../su99/vogel/hoodsketch.jpg
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I
III
BSC Class
Protection
Sample Χ
Operator √
Environment √
Application with Chemicals
Nonvolatile Toxic ChemRadionuclide
√
Volatile ToxicRadionuclide
√*
Protection
Sample √
Operator √
Environment √
Application with Chemicals
Nonvolatile Toxic Chem Radionuclide
√
Volatile Toxic ChemRadionuclide
√ (Small
amount)
Airflow patternIn at front
Exhausted through HEPA to the outside* or into the room.
Airflow patternSupply air inlets and hard-duct exhausted to outside through two HEPA filters in series.
BMBL 5th
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II A1Protection
Sample √
Operator √
Environment √
Application with Chemicals
Nonvolatile Toxic Chem Radionuclide
√
Volatile Toxic ChemRadionuclide
Χ
Airflow pattern70% recirculated to the cabinet work area through HEPA
Exhaust air recirculate to the laboratory or discharged from the building via a canopy connection.
II A2 (A/B3)Protection
Sample √
Operator √
Environment √
Application with Chemicals
Nonvolatile Toxic ChemRadionuclide
√
Volatile Toxic ChemRadionuclide
√* (minute amounts
)
Airflow patternSame as II, A1
Exhaust air recirculate to the laboratory or discharged from the building via a canopy connection*.
BSC Class
BMBL 5th
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Biosafety Level 2: Safety Equipment (Primary Barriers and Personal Protective Equipment)
• Remove protective clothing before leaving for non-laboratory
• Dispose of or deposit for laundering protective clothing appropriately.
• Persons who wear contact lenses in laboratories should wear eye protection.
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Biosafety Level 2: Safety Equipment (Primary Barriers and Personal Protective Equipment)
• Select glove base on an appropriate risk assessment. Gloves must not be worn outside the laboratory.
Remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory.
Do not wash or reuse disposable gloves.
Dispose of used gloves with other contaminated laboratory waste.
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Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
• Laboratory doors should be self-closing and have locks.
• Laboratories must have a sink for hand washing.
• The laboratory should be designed so that it can be easily cleaned and decontaminated. Carpets and rugs in laboratories are not permitted.
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• Laboratory furniture must be capable of supporting anticipated loads and uses.
• Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals.
• Chairs used in laboratory work must be covered with a non-porous material.
Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
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• Laboratory windows that open to the exterior are not recommended. Otherwise, must be fitted with screens.
• Vacuum lines should be protected with HEPA filters, or their equivalent. Liquid disinfectant traps may be required.
Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
flickr.com/photos/70136696%40N00...84133289
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• BSCs must be installed so that fluctuations of the room air supply and exhaust do not interfere.
Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
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• An eyewash station must be readily available.
• Mechanical ventilation systems provide an inward flow of air without recirculation to spaces outside of the laboratory.
Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
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• HEPA filtered exhaust air from a Class II BSC can be safely re-circulated back into the laboratory environment if the cabinet is tested and certified at least annually.
Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
web.princeton.edu/.../labsafetyman ual/bsc1 .jpg
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• A method for decontaminating all laboratory wastes should be available in the facility
(e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method).
Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
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Disinfectant
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Autoclave
What should not be autoclaved?• Corrosive chemicals• Flammable chemicals• Combustible• Explosive• Radioactive materials
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http://www.phac-aspc.gc.ca/msds-ftss/index-eng.php#s
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Animal Biosafty Level 2
Standard Microbiological PracticesSpecial PracticesSafety EquipmentLaboratory Facilities
Safe Practice
Facility Design
Safety Equipment
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Animal Biosafety Level 2: Standard Microbiological Practices
• Prior to beginning a study animal protocols must be reviewed and approved by the IACUC and the Institutional Biosafety Committee.
• A safety manual specific to the animal facility is prepared or adopted
• The need for an animal allergy prevention program should be considered.
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• The animal facility is separated from areas that are open to unrestricted personnel traffic within the building.
• External facility doors are self-closing and self-locking.
• Doors to areas where infectious materials and/or animals are housed, open inward, are self-closing.
Animal Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
www.dearborn-animals.com/New%252...plan.htm
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Animal Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
• Sink traps are filled with water, and/or appropriate liquid to prevent the migration of vermin and gases.
• Floor drains must be maintained and filled with water, and/or appropriate disinfectant to prevent the migration of vermin and gases.
home.howstuffworks.com/home-impr...ains.htm
www.diychatroom.com/f7/do-we-hav.../index3/
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Animal Biosafety Level 2: Laboratory Facilities (Secondary Barriers)
• External windows are not recommended; if present, windows should be sealed and must be resistant to breakage.
• Cages should be autoclaved or otherwise decontaminated prior to washing. Mechanical cage washer should have a final rinse temperature of at least 180°F.
www.detach.se/products%2520-%252...hing.htm
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References
• American Biosafety Association http://www.absa.org/riskgroups/index.html.
• CDC-NIH, 2007. Biosafety in Microbiological and Biomedical Laboratories (Fifth Edition).
• WHO, 2004. Laboratory Biosafety Manual.
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Thank You and Questions?Thank You and Questions?