Biosafety, Biosecurity and PAPR Annual Training

Safety, Security & PAPR Annual Training 2016 North Carolina State Laboratory of Public Health Bioterrorism and Emerging Pathogens Unit *Required for new employees, annual refresher and as needed when changes occur Biosafety Biosafety is the application of combinations of laboratory practice and procedure, laboratory facilities, and safety equipment when working with potentially infectious microorganisms. Biosafety guidelines are put into place to protect laboratorians and prevent exposures in biological laboratories.

Everyone working in the laboratory is responsible for employing proper biosafety practices at all times and in all situations. Principles of Biosafety Good biosafety practices prevent intercepting microorganisms. occupationally acquired infections by

Blocking the routes of transmission by intercepting microorganisms before they can infect through proper work practices, protective equipment, immunization, treatment and surveillance Reservoir of pathogen Portal of escape Transmission Route of entry/infectious dose Susceptible host Incubation period Infection Biosafety Plan Key Elements Biosafety manual and General laboratory

SOPs Responsibilities Identification of biohazards Entry requirements and signage Risk assessments and control of biohazards Packaging and shipping protocols safety practices Waste management Decontamination/Disinfection Emergency procedures Training program

Medical surveillance Evaluation/Drills/ Auditing program and Documentation Containment Containment (barrier) describes safe methods, facilities and equipment for managing infectious materials in the laboratory environment where they are being handled or maintained. Purpose: Manage infectious materials Reduce or eliminate exposure

Elements: Safety Equipment (Primary Containment) Facility Design and Construction (Secondary Containment) Laboratory Practice and Technique The most important element of containment is strict adherence to standard microbiological practices and techniques. Standard practices and procedures Common to all laboratories, handling sharps, decontamination Special practices and procedures Entry and Exit procedures, Address the risk of handling agents requiring increasing levels of

containment Administrative- Immunizations, security clearance, training and supervision Standard Microbiological Practices and Techniques Limited or restricted access to the laboratory Hand washing protocols No eating, drinking, smoking, handling contact lenses or applying cosmetics in the laboratory Use of mechanical pipetting devices No mouth pipetting! Waste decontamination and disposal program

Insect and rodent control program Minimization of splashes and aerosols Procedures That Emit Aerosols Catalase Inoculating biochemicals or blood culture bottles Pipetting Mixing Centrifugation Grinding Vortexing Pouring Loading syringes

Lasers, cell sorters Splashes Opening lyophilized cultures Flaming loops Entering or opening vessels at non-ambient pressures, fermenters, freezer vials Ways to Minimize Aerosols Pour liquids carefully Work over absorbent

bench pads Use centrifuge safety cups Use sealed centrifuge rotors Use capped tubes when mixing or vortexing Use pipette aids with filters Work in BSC Primary Containment (Barriers) Primary containment is provided by the use of appropriate safety equipment. It provides protection of personnel and the immediate

laboratory environment from exposure to infectious agents. Biosafety Cabinet Eye/Face Protection Safety Centrifuge Cup Lab Coat/Gown Respiratory Protection Gloves Shoe covers

Biosafety Cabinet The Biosafety Cabinet is the principal device used to provide containment of infectious splashes or aerosols generated by many microbiological procedures. Secondary Containment Facility Design and Construction The design and construction of the facility contributes to the laboratory workers protection, provides a barrier to protect persons

outside the laboratory, and protects persons or animals in the community from infectious agents which may be accidentally released from the laboratory. Facility Design and Construction Separation of the laboratory work area from public access Availability of a decontamination facility (autoclave) Hand washing facilities Specialized ventilation systems to ensure directional airflow Air treatment systems to decontaminate or remove agents from exhaust air

Controlled access zones Airlocks at laboratory entrances Separate buildings or modules to isolate the laboratory Biosafety Levels Biosafety Levels Laboratories are divided into four different safety levels that basically determine the type of work that can be performed in that laboratory. Provide increasing levels of personnel and environmental protection Consist of combinations of laboratory practices and techniques, safety equipment and laboratory facilities The performance of risk assessments is necessary and

mandatory. Generally, work with known agents should be conducted at the biosafety level recommended in the BMBL Section VII When specific information is available to suggest that virulence, pathogenicity, antibiotic resistance patterns, vaccine and treatment availability, or other factors are significantly altered, stringent practices may be specified BSL 1 Very Basic Used when handling agents not known to cause disease in healthy humans No primary barriers

required Open bench top Hand washing sink Lab coats and gloves Sharps policy BSL 1 BSL 2 Lab activities involving clinical specimens, blood, other body fluids and tissues from humans or animals potentially infected with human

pathogens. Lab coats and gloves at minimum Hand washing sinks Waste decontamination facilities Risk of percutaneous injury, ingestion, mucous membrane exposure Medical surveillance policy BSC or safety centrifuge cups

BSL 3 Used for lab activities involving work with indigenous or exotic agents which may cause serious or potentially lethal disease as a result of exposure by the More emphasis placed on primary and secondary barriers to protect personnel, the community and the environment from exposure to potentially

BSL 3 BSL-3 Primary Barriers Class I or Class II BSCs or other physical containment devices used for all open manipulations of agents PPE - protective lab clothing, gloves, goggles, respiratory protection as required by risk assessment (PAPRs) Secondary Barriers Engineering controls Physical separation from access corridors Self-closing , double-door access doors HEPA filtration system Exhausted air not recirculated Negative airflow into laboratory Decontamination of ALL waste

Eyewash available Raleigh BSL-3 Facility BSL-3 Entry and Exit Procedure Visitors Log PPE Requirements Donning and Doffing of PPE Transfer of Biohazardous Agents into the Laboratory Precautions for Sharps Gross Decontamination HVAC, Alarms, BSC, Local and Air Pressure Differential Alarms HEPA Filter Electrical System

BSL-4 Dangerous/exotic agents which pose high risk of life threatening disease, aerosol-transmitted lab infections; or related agents with unknown risk of transmission Primary hazards to personnel are respiratory exposure to infectious aerosols, mucous membrane or broken skin exposure to infectious droplets or auto inoculations Two types of BSL-4 laboratories: Cabinet = all procedures conducted in Class III BSC Suit = all procedures conducted in Class I or II BSC in combination with full-body, air supplied pressure personnel suit

Laboratory Risk Assessments Risk Assessments Risk assessment is a process used to identify: the hazardous characteristics of a known infectious or potentially infectious agent or material the activities that can result in a persons exposure to an agent the likelihood that such exposure will cause a lab acquired infection the probable consequences of such infection A risk assessment involves trying to predict what might go wrong, how likely it is to go wrong and how severe the consequences would be. Risk = the chance of injury, damage, or loss Chance = the probability of something happening Hazard = something that is dangerous A risk assessment must be completed for each organism that is used in the

laboratory. When to Perform Laboratory Risk Assessments At regular intervals - at least annually Laboratory move or renovation New employee New infectious agent or new reagent New piece of equipment Changes in protocols/procedures Components considered in developing a risk assessment Men Qualification Experience and Training

Attitude Immune status Physical handicaps Materials Pathogens and toxins Pathogenicity Mode of transmission Reagents Containers and Four Ms Machinery

Building Laboratory design Facilities Air flow (air handling) Equipment BSC, Fume hoods Centrifuge cups/carriers Methods Procedures SOPs Documentation According to the BMBL There are things to be considered after an organism or

method has been chosen. These items should be included in a biological risk assessment: Pathogenicity disease incidence and severity, LAIs Route of transmission inhalation, ingestion, cutaneous Agent Stability survival over time in the environment Infectious dose how much? Immune status? Concentration working volume, # of infectious organisms per unit Origin domestic or foreign in origin. Host? Availability of prophylaxis/therapeutic treatment vaccines, antibiotics and anti-viral meds Medical Surveillance Serum banking, monitoring employee health status, post-exposure management Experience and skill level of personnel laboratorians, maintenance, housekeeping

Useful Tools for Performing Risk Assessments Review laboratory records Injury, illness and surveillance reports Equipment maintenance records (is the equipment well maintained/inspected/certified) Inspect the laboratory Daily monitoring by employees, periodic walk-throughs and formal inspections

Review published materials Equipment manuals, manufacturers bulletins, package inserts, scientific journals, safety manuals and guidelines Observe laboratory operations New procedures, new employees, new equipment, workflow Controls to Reduce Risk Avoid the hazard Do not perform the assay or handle the agent Use alternates/surrogate organisms Procedural Controls Perform assay when less people are around Separate areas Engineering/Mechanical controls

BSC Mechanical pipetting devices PPE Determine specific types(s) of PPE required (i.e. respirators) Emergency Procedures Incident/Accident response procedures Health Surveillance Benefits of Completing a Risk Assessment Effective use of resources Identification of training needs and supervision Advance planning for renovation Evaluation of procedural changes Prevention of biohazard transmission to family

members of employees Ensure compliance with governmental regulations Justification for space and equipment needs Cost effective laboratory operation Evaluation of emergency plans Entry and Exit Procedure BSL-3 Entry and Exit Procedures are posted at the main entrance and to each individual BSL-3 suite. Laboratory Entry Notify team mate that you will be working in the BSL-3

laboratory. Access using security measures access key card, biometrics When scanning for entry, all employees wishing to enter the lab must scan their card key/fingerprint. Observe any signage on anteroom door maintenance, do not enter, etc. Document entry in log book all visitors must read the precautions document and sign the Visitors log. Document that the visual airflow device or indicator was checked and that air is flowing properly. Don PPE located in anteroom Observe and post proper sign on individual BSL-3 suite door includes occupancy, types of PPE in use, maintenance, etc. Potential Risks and Hazards for Visitors

Minimum PPE Requirements while infectious agents are not in use: i.e. Taking Temperatures, Inventory, Restocking, Maintenance Personnel, Tours, etc. Donning order while in Sign-in Room: Bootie/shoe covers Gown unless you are re-using a gown stored in the common area Gloves, single layer - Required for entry; this layer of gloves should be considered hands and must be worn at all times when in the BSL-3 Protective eyewear Minimum PPE Requirements for work with infectious agents:

Donning order while in Sign-in room: Bootie/shoe covers Jumpsuit, if required by risk assessment Gown - unless you are re-using a gown stored in the common area Gloves, single layer - Required for entry; this layer of gloves should be considered hands and must be worn at all times when in the BSL-3 Protective eyewear Donning order while in the BSL-3 Common Area: For all testing requiring the use of PAPRs, the PAPR will be donned in the common area before entering the BSL-3 corridor Donning order while in the BSL-3 Corridor: Second pair of gloves - Required when working with agents Personal Protective Equipment

Doffing PPE Doffing order in isolation room: Discard sleeves, if worn, and outer layer of gloves while in BSC Remove gown if jumpsuit is also worn Doffing order in BSL-3 Corridor: Doff jumpsuit, if worn, and discard into biohazard bag If no jumpsuit is worn, Doff gown and discard into biohazard bag Remove and discard booties into biohazard bag Doff the inner layer of gloves Doffing order in common area: Don clean gloves to doff PAPR unit Doff PAPR unit. Place hood in biohazard bag and place PAPR unit in a holding bin. If re- entry is required, use a new hood.

If sample is negative, store one hood per laboratorian and recharge PAPR unit for reuse. If sample is positive, decontaminate PAPR unit(s) and recharge for reuse. If breach is suspected, discard hood and change filter. Remove gloves and discard into a biohazard bag. Wash hands. Doffing PPE NOTE: In accordance with the NCSLPH Exposure Control Plan: all personal protective equipment is removed immediately, or as soon as possible, if overtly contaminated and placed in an appropriately designated area or container for storage, washing, decontamination, or disposal. This includes disposal or decontamination of face and eye protection prior to reuse. Laboratory Exit

Properly dispose of PPE Change sign Wash hands Exit to anteroom Complete logbook Exit Powered Air Purifying Respirator (PAPR) PAPR Motor blower draws contaminated air through a HEPA filter and blows filtered air up into the head covering. The PAPR provides respiratory protection again airborne contaminants:

Dust, fumes, mists, smoke PAPRs DO NOT reduce exposure to gases or vapors Note the difference between fumes and vapors fumes - heated solid particles temporarily suspended in air vapors - gaseous The PAPR with the head cover reduces exposure by a factor of 25, PAPR with hood reduces exposure by a factor of 1000. PAPR Prior to wearing a PAPR, staff must undergo medical evaluation. The facilitys Respiratory

Protection Program Director reviews the medical clearance questionnaire and decides if employee needs to be referred to a personal physician for clearance to wear a PAPR. Physiological stresses Pulmonary Cardiac Signs and symptoms limiting use Claustrophobia, latex allergy PAPR components Hood Blower / Filtration Unit (w/Airflow

Indicator) Breathing Tube Battery Charger Preparing to Wear the PAPR 1. Inspect PAPR unit. Replace if cracked or warped. 2. Check breathing tube for any cracks or tears. The tube should be flexible. Pay particular attention to the rubber O-ring gasket replace if it is worn, cracked or no longer flexible. 3. Attach the breathing tube to the

blower/filter/battery unit by inserting the male end of the hose and turning it clockwise until it stops. 4. Turn the power on. 5. Check the airflow with the airflow indicator. Check Airflow with the Airflow Indicator 1. 2. 3. 4. 5. Turn the PAPR power on. Place the indicator in end of the hose.

The indicator should float on the air coming out of the tube. Two lines on the indicator should be visible. If the test fails, do not use the unit. Perform troubleshooting. Problem Possible Causes Low airflow

Battery needs changing Filter is loaded PAPR blower malfunction Breathing tube restricted Corrective Action

Switch to fully charged battery Replace filter Switch to a different blower Remove restriction Changing a PAPR Filter The PAPR filter and gasket should be replaced when: 1.

2. 3. 4. 5. 6. PAPR doesnt pass airflow check Filter is physically damaged Water has entered the filter To comply with administrative procedures Gasket is torn or damaged A breach is suspected Changing a PAPR Filter

1. 2. 3. 4. 5. 6. Release back cover locking tabs and remove the back cover. Remove the filter and filter gasket. Autoclave and dispose of the filter and gasket. Remove the battery pack. Inspect the battery and the housing. Replace if damaged, cracked or no longer holds a charge. Insert new filter and filter gasket.

Replace back cover. Test PAPR using airflow indicator. PAPR Donning After all other appropriate PPE has been donned: 1. Obtain PAPR components and hood. 2. Attach PAPR to the waist, latching in the front. 3. Insert breathing tube into the PAPR unit. Twist 4. 5. 6. 7. 8. breathing tube/air hose to secure into the unit. Turn on PAPR unit.

Attach PAPR breathing tube to the hood. Place hood over head and shoulders. Tuck inner skirt into the jumpsuit. Put the gown on last. PAPR Doffing After all appropriate levels of PPE have been doffed: 1. 2. 3. 4. 5. Remove hood from head. Turn off PAPR unit.

Remove breathing tube from PAPR unit. Remove PAPR unit from waist. Place tube and PAPR unit in designated area for re-processing. Limitations Impaired lung function Communication and vision problems Fatigue Reduced work efficiency Claustrophobia Battery failure Only in atmospheres with sufficient O 2 Over breathing Latex allergies

Risks Improper use training must be provided and documented annually for each user. Breakthrough Contaminants pass through the filtering material when organic vapors and gases cause the filter to reach capacity. Penetration improper seal is created allowing aerosols or other harmful air contaminants to enter. Degradation of effectiveness occurs when parts of the PAPR begin to deteriorate. Contamination of wearers when doffing may occur if the PAPR is doffed improperly. PAPR

Cleaning/Disinfection/Storage Cleaning Wipe the outside surfaces with a mild solution of warm water and mild detergent. Do not clean with organic solvents. If necessary, wipe with a 10% bleach solution. PAPR hoods are disposable, but may be reused by a single user and must be cleaned between each use. Disinfection/Storage Disinfect the PAPR after each use 10% bleach solution Battery pack should be stored fully charged Store PAPR so that it is protected from damage, contamination, dust, sunlight, extreme temperatures, excessive moisture, and damaging chemicals.

Store PAPR in a cool, dry place. General Transfer of Biohazardous Agents into a High-Security Laboratory No infectious agents should be transported via the public elevator. Use the freight or specimen elevator for the transport of samples. Specimens of a suspected bioterrorism event should remain un-opened until they are inside the biological safety cabinet dedicated for this task. Infectious agents should be placed into an unbreakable, secondary container or double-bagged in a

leak proof container, both of which are sealed tightly with O-ring seal. All containers of size 250 ml or greater should be transported on a cart or in a bottle carrier. Precautions for Sharps Disposable sharps are placed in sharps containers that are autoclaved when the container is two thirds full. Disposable sharps are not to placed in the reusable waste containers with bag inserts. Non-disposable sharps are placed in a hard walled container such as a metal tray for transport to the autoclave and sterilized before re-use.

Sharps involved in molecular testing should not be re-used due to possible nucleic acid contamination. Gross Decontamination The appropriate method of decontamination will be based on the organisms and quantities in use. Ports in the walls of individual rooms to facilitate gross decontamination using Vaporized Hydrogen Peroxide (VHP). If gross decontamination were to occur in a BSC or BSL-3 room, perform VHP decontamination of those spaces. Refer to the risk assessment to determine the proper decontamination method.

During decontamination, the laboratory or designated spaces may be restricted to all investigators. BSCs and Local Alarms To ensure that local alarms sound in each isolation room in the BSL-3 when there are either airflow disruptions or exhaust system failures, BSCs will remain ON at all times. Responding to HVAC alarms In the case of airflow disruption or exhaust system failure: Audible alarms on actively operating Biosafety Cabinets will sound

Air pressure differential alarms, both audible and visual, in individual rooms will alarm if the pressure differential becomes positive. Upon hearing the alarms, the laboratorians should assume that negative pressure has been compromised and immediately secure any biological agent being worked upon Leave the BSL-3 following proper procedures and post the DO NOT ENTER signage on the anteroom door. The Responsible Official, BTEP supervisor, Laboratory Director, and appropriate maintenance personnel should be notified of the situation as soon as possible. After hours, the state capital police should be notified 919-733-3333 and they will contact the afterhours DOA maintenance administrator. Air Pressure Differential Alarms The BSL-3 laboratory is designed for 100% single

passage, uni-directional flow-through air. The exhaust air system is electronically connected to the supply air system. The fans of these systems are programmed to maintain a predetermined cfm (cubic feet per minute) flow differential to maintain the negative pressure in the BSL-3 suite in relation to the adjacent lab areas. Visual and audible alarms will indicate if the differential nears neutral or reaches positive pressure. If activities occur that will knowingly sound the alarm (e.g. opening the door, etc.), staff will alert other team members working in the area of the impending alarm. Otherwise, all

work should safely cease and alarm investigated/assessed/documented/reported. HEPA Filters Suite Fans Filter Bank Modules Unknowns EF 3-1. 3-2 F-3*, **

EF5-1. EF5BT/Virology 2 F-2 1 9 Each BSL3 lab suite has two exhaust bag-in/bag-out HEPA filter housings and two exhaust fans which are located in the mechanical penthouses and roof. Each filter bank has pre-filters and HEPA filters. Some filters also include HEGA (carbon filtration for chemicals) filters (this varies with each suite). The supply is not HEPA filtered. *This suite does not have redundant filter banks. ** This suite also has HEGA filters inline with the HEPA filters. Electrical System Emergency Generator restores power within 10 seconds All BSCs have UPS unit to bridge gap of time

ROOM NUMBERS 3916 is panel # OL3A5 3921 is panel # OL3A5 3922 is panel # OL3A5 3923 is panel # OL3A5 and and and and circuit circuit circuit circuit

# # # # 29 41 17 & 3 (2 cabinets in the room) 9 & 11(2 cabinets in the room) 3903 is 3904 is 3905 is 3906 is 3907 is

and and and and and circuit circuit circuit circuit circuit # # # #

# 27 38 33 19 1 panel panel panel panel panel # #

# # # OL3A6 OL3A6 OL3A6 OL3A6 OL3A6 The cabinets are being fed from circuits OL3A5 and OL3A6. These 2 panels are themselves being fed from the distribution panel # ODL3A1 in the electrical room #3136. ODL3A1 in turn is fed from transformer #T7 in room #3136 and the transformer is fed off of switchboard A. Emergencies in the

BSL-3 Emergencies in the BSL-3 Personal safety comes first! Personnel shall not expose themselves to any unnecessary risks in order to comply with select agent or BSL-3 policies. Fire Natural Disaster Hurricane Tornado Equipment failure PAPR BSC

Loss of power Interruption of air flow Emergencies in the BSL-3 Upon hearing any emergency alarm: 1. Use common sense! 2. Secure any organisms to the best of the investigators ability prior to leaving the BSL-3. 3. Remove as much PPE as possible without selfcontamination or self-endangerment and wash your hands. *During a fire alarm, the interlock will be engaged. This will not allow both doors to be open. If bypass is needed, press the green over-ride button next to the door. Alternative exit emergency methods include doors in equipment room (3902 & 3919).

BSL-3 Laboratory Emergencies Emergency Response Evacuation Route Dial 8-911 Pull fire alarm located in hallway Building operator: 37834 Other numbers: 919733-3333 (State Capitol Police 24-hour Law Enforcement presence on-site)

Exit BSL-3 lab suite via exterior door. Turn right and exit building via closest stairwell. Upon exiting building, assemble in the third row of the front parking lot. Check in with supervisor. Biosecurity Security Measures Entry/exit reports Access proximity card Biometrics

Video surveillance Motion detectors Locked incubators, freezers, and refrigerators Knox box 24-hour law enforcement presence Other security measures Security risk assessments Inspection of all packages to reduce the risk of

an unauthorized removal of agents or information from the laboratory. Security plan describes how the institution will handle unacceptable risks. A chain-ofcommand and roles and responsibilities must be clearly defined. Ensure that the plans are created, exercised and revised as needed. Notify your Responsible Official Security related incidents may occur that will warrant the notification of the Responsible Official: Loss of keys Termination of employment Loss, theft, or release of Select Agent Alteration of inventory records

Unauthorized entry to the laboratory Symptoms consistent with lab-acquired infections Most common Brucellosis and Tularemia Responsible Parties Controller Lou Turner Responsible Official Dee Pettit

Alternate Responsible Official/ Safety Officer Kristy Osterhout Incident Response Form Nature of threat Breach discovery i.e. potential exposure Details of occupational release Agent or toxin missing Actions taken to resolve the incident Determination of cause Preventive actions Evaluation of new measures

References http://www.cdc.gov/biosafety/publications/bmbl5/ http://www.selectagents.gov/resources.html http:// multimedia.3m.com/mws/media/735385O/3m-air-mate-h igh-efficiency-powered-air-purifying-respirator-papr-use r-instructions.pdf

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