BIOSAFETY LEVEL 3 (BSL-3) FACILITYVA Long Beach Healthcare System

Updated 06/02/16

TB Laboratory Safety PlanEmergency Response Plan

BSL-3 Security PlanStandard Operating Procedure

Stephanie Alley, MA, SRS Chair Date

Timothy R. Morgan, M.D., Chair R&D Date

Chris Reist, M.D., ACOS/R&D Date

Michael W. Fisher, Medical Center Director Date

1

Table of Contents

INTRODUCTION........................................................................................................................................... 4

I. Responsibilities....................................................................................................................................... 4

A. Laboratory Director............................................................................................................................. 4B. Safety Committee................................................................................................................................ 4C. Research Personnel.............................................................................................................................. 4D. The VA Occupational Health Clinic....................................................................................................4

II. Medical Surveillance Program................................................................................................................. 5

A. Purpose................................................................................................................................................ 5B. Purified Protein Derivative Monitoring................................................................................................5C. Other Pathogens................................................................................................................................... 5D. Respirator Fit Testing and Medical Approval.......................................................................................5

III. Facility and Equipment........................................................................................................................ 5

A. Facility Rooms.................................................................................................................................... 6B. Door Locking System.......................................................................................................................... 6C. Air Balancing System.......................................................................................................................... 6D. Biosafety Cabinets............................................................................................................................... 6E. Autoclave............................................................................................................................................ 6F. Other equipment.................................................................................................................................. 7

IV. Maintenance and Repair Personnel, Contractors, and Visitors..............................................................7

A. Entering the BSL-3 Facility................................................................................................................. 7B. Responsibilities of the BSL-3 Facility..................................................................................................7C. Responsibilities of Visitors to the BSL-3 Facility.................................................................................7

V. Standard Operating Procedures................................................................................................................ 7

A. BSL-3 Entry Authorization.................................................................................................................. 7B. Protocol Submission............................................................................................................................ 8C. Personnel Training............................................................................................................................... 8D. Work Practice Controls........................................................................................................................ 9E. Entry Preparation................................................................................................................................. 9F. Exit Procedures.................................................................................................................................. 10

VI. General Lab Procedures..................................................................................................................... 10

A. Biological Safety Cabinets................................................................................................................. 10B. Shipping and Receiving Specimens....................................................................................................11C. Working with Organisms................................................................................................................... 12D. Specimen Transporting/Removal from Facility..................................................................................12E. Specimen Storage.............................................................................................................................. 12F. Biohazard Waste................................................................................................................................ 12G. Autoclaving Waste in the Facility......................................................................................................13H. Radiation Use.................................................................................................................................... 13I. Facility Maintenance......................................................................................................................... 13

2

VII. Emergency Procedures....................................................................................................................... 14

A. The BSL-3 Facility Air-Flow System.................................................................................................14B. Spills................................................................................................................................................. 14C. Exposure Incidents............................................................................................................................. 15D. Power Failure.................................................................................................................................... 15E. Fire................................................................................................................................................... 15 F. Earthquake......................................................................................................................................... 16

Appendix 1 - BSL-3 Facility ........................................................................................................................ 17Appendix 2 - Packaging and Labeling of Etiologic Agents............................................................................18Appendix 3 - Respirator Program.................................................................................................................. 25Appendix 4 - Emergency Distribution List....................................................................................................28Appendix 5 - Incident Report Form...............................................................................................................29

Attachment A................................................................................................................................................ 30Attachment B................................................................................................................................................ 31References.................................................................................................................................................... 32

3

BIOSAFETY LEVEL 3 (BSL-3) FACILITYVA Long Beach Healthcare System

INTRODUCTIONThis manual has been prepared for researchers using the Biosafety Level 3 (BSL-3) Facility located at the VA Long Beach Healthcare System (LBVAHS), Building 138. Use of the Facility is limited to researchers who comply with the "authorized user" requirements described in subsection D below.I. Responsibilities

A. Laboratory Director – Charles Sohaskey Ph.D.

The laboratory director exercises general administrative responsibility for the Facility. He has the responsibility and authority to perform risk assessments of projects, establish policies and procedures, train personnel, and ensure maintenance of the Facility and equipment. The laboratory director is responsible for ensuring that the personnel apply safe practices and procedures at all times while working at the BSL-3 Facility. He is ultimately responsible for who enters the BSL-3.

During the conduct of research, the director is responsible for

supervising the safety performance of the staff to ensure that required safety practices and techniques are employed, and

investigating and reporting in writing to the Subcommittee on Research Safety any accidents or problems pertaining to potential exposure hazards, the compromise of the biological and physical barriers, or equipment or Facility failure, and correcting procedures that may result in hazardous spill and overt personnel contamination.

B. Subcommittee on Research Safety

The Subcommittee on Research Safety establishes operational rules regarding safety and use of the facility. The Committee reviews and approves proposed projects using the BSL-3. The Committee can suspend access privileges of any person(s) found to be in violation of the policies and procedures governing the use of the Facility.

C. Research Personnel

Research personnel must comply with the following to conduct research within the BSL-3 Facility:

Follow all procedures outlined in this BSL-3 Manual

Learn the laboratory operational and emergency procedures, understand the potential hazards of the infectious agents in use, and help maintain the Facility in good working condition

Report all exposures or illnesses related to the Facility to the Lab Director

Have completed medical surveillance and training requirements

Perform assigned responsibilities

D. The VA Occupational Health Clinic

The VA Occupational Health Clinic will provide the following services to both VA employees and non-VA employees designated as workers without compensation (WOCs): Purified Protein Derivative (PPD) testing

Post-exposure prophylaxis for exposure incidents

4

Exposure reporting

Counseling

II. Medical Surveillance Program

A. Purpose

A medical surveillance program for personnel using the BSL-3 Facility is conducted by the VA Occupational Health Clinic at no additional charge. This will include having a PPD test done prior to conducting work in the Facility.

It is the responsibility of the PI to ensure new employees are tested at the VA Occupational Health Clinic.

B. Purified Protein Derivative Monitoring

The VA Occupational Health Clinic will administer the PPD intradermal skin test (Mantoux test) when work in the BSL-3 Facility is started (assuming a previous negative PPD) and every year thereafter. A positive skin test indicates that an individual has been exposed to either M. tuberculosis or M. bovis (including BCG the vaccine strain). If an individual has had a previous positive PPD test they will document an annual respiratory self assessment to check for symptoms of tuberculosis.

Previous history of PPD positive individuals will be noted. They will be counseled on various options of health care, such as chest x-rays or preventative drug treatment. If an employee previously shown to be PPD negative subsequently shows a positive result, the Subcommittee on Research Safety must be notified and an evaluation performed. The individual's techniques should be reviewed for possible breaches in safety and the BSL-3 Facilities' safety procedures will be re-evaluated.

C. Other Pathogens

Handling blood samples carries the risk of being exposed to diseases, particularly hepatitis and HIV. Personnel who handle blood must be identified by their department as at risk, be included in a bloodborne pathogens program (BBP), and offered BBP training and the hepatitis B vaccine, through the VA Occupational Health Clinic and Infection Control.

All employees should be made aware of the dangerous link between tuberculosis and HIV infection due to compromised immune response. HIV positive people who become infected with tuberculosis are much more likely to develop active disease. As HIV status is a confidential matter, each employee must assess his or her own situation. The HIV Program coordinator or HIV Program Director may be consulted for further guidance.

D. Respirator Fit Testing and Medical Approval

On an annual basis the VA Safety Office will perform N95 respirator fit testing and required respirator training for all individuals working in BSL-3 Facility.

III. Facility and EquipmentAs stated by the National Institutes of Health (NIH) and the Centers for Disease Control and Prevention (CDC) in the 4th edition of the Biosafety in Microbiological and Biomedical Laboratories, certain types of work with Mycobacterium tuberculosis require a BSL-3 environment. A research facility must meet certain minimum specifications. The TB laboratory room 207 located in Building 138 at the LBVAHS has been designed and constructed to meet the requirements work with live cultures of Mycobacterium tuberculosis and other organisms requiring similar containment. A biohazard sign is posted on the door to warn individuals of entry restrictions.

5

A. Facility Rooms

The BSL-3 Facility is comprised of three rooms. Please refer to Appendix 1 for a Facility diagram.

The ante-room is a safe zone where the operator can gown-up in preparation for entry into the inner rooms, where the infectious work is done.

Inner Room

Hood room with biosafety cabinets

B. Door Locking System

The outer door must be locked at all times. To prevent sudden changes in negative room pressure, the outer door must be closed before the inner door is opened.

C. Air Balancing System

M. tuberculosis can be carried on water droplets small enough to float in the air, but they can be removed by directed air currents. The BSL-3 laboratory has negative air pressure (minimum 0.001 inches of water) that maintains an airflow direction from the outside hallway, into the ante-room, then into the inner room and finally through HEPA filters with a ventilation rate of at least 12 ACH (Air Changes per Hour). Maintaining a negative air pressure gradient is essential for safe operation.

The air balancing system is designed to maintain a negative air pressure gradient and to provide an air exchange frequency of at least twelve air exchanges per hour. LBVAHS facilities personnel will monitor the air balance readings annually as part of routine maintenance. Negative pressure is verified by one audible and two visual alarm systems.

There are 2 monitoring devices for the BSL-3. The main system with audible alarm has sensors in the inner room and the hallway and is mounted in the back of the inner room. The other system is mounted in the ante-room and can be seen from the hallway. A red light on the monitor indicates a failure of the system, and that the room is under positive pressure. A red light on the separate box (with green light on the main monitor) indicates the backup system was activated to prevent loss of negative pressure. If the lab has lost negative pressure then Charles Sohaskey should be notified immediately.

The air is vented to the outside after being filtered through HEPA filters in the room air ducts. The performance of the HEPA filters will be monitored annually by a contractor as specified in "Class II Laminar Flow Biohazard Cabinetry, NSF International Standard 49-1992."

The air pressure in the lab is recorded weekly on the Lab Event Sheet.

D. Biosafety Cabinets

The Hood Room contains two Class II biological safety cabinets. After use the inside surface is cleaned and the UV light source is turned on. The UV fixture must be left on for a minimum of 20 minutes between users. Remember not to look directly at the UV fixture. LBVAHS facilities personnel are responsible for maintaining UV fixtures.

The biosafety cabinets will be monitored every six months for proper airflow, UV light, and HEPA filter performance according to NSF Standard 49. NIH/CDC guidelines specify the certification of biosafety cabinets initially, upon disruptive activities, and at least annually thereafter.

E. Autoclave

There is one autoclave in the Facility. BSL-3 Facility personnel must autoclave all infectious waste for an appropriate cycle time. Personnel will place containers of waste for autoclaving on the cart next to the autoclave. To ensure proper sterilization, a spore test will be performed by BSL-3 personnel. The results are recorded on the Lab Event Sheet.

6

F. Other equipment

The BSL-3 also contains an eye wash that should be checked by lab personnel every week. To test the lever is pressed and water allowed to flow for ~20 sec (until water runs clear). The result is recorded on the Lab Event Sheet. A fire extinguisher is the anteroom. This is examined every month by the VA and documented.

IV. Maintenance and Repair Personnel, Contractors, and VisitorsNo unauthorized users are allowed in the BSL-3 without prior notification being given to the Lab Director. All maintenance or repair personnel, contractors, and visitors must first be apprised of the possible risks associated with the Facility, and be willing to perform work in the Facility under these circumstances described.

A. Entering the BSL-3 Facility

Visitors are all individuals who do not work in the BSL-3 Facility, including, but not limited to, VA maintenance personnel, contractors, outside consultants or visitors, etc.

All visitors shall follow the Facility’s entry requirements with respect to personal protective equipment (PPE) use.

All laboratory worker activities will be suspended and appropriate surfaces decontaminated prior to entry by VA maintenance personnel and outside contractors. Any equipment requiring repairs must be decontaminated prior to allowing access to the unit.

B. Responsibilities of the BSL-3 Facility

Discontinue all research activities/experiments within the BSL-3 for the duration of the repair and/or maintenance work.

Individuals who have been trained to work in the Facility shall escort all visitors into the BSL-3 Facility.

Provide all necessary PPE for entering the BSL-3 Facility.

C. Responsibilities of Visitors to the BSL-3 Facility

Contact designated personnel and schedule times when work can be done.

Follow all posted entry/exit procedures.

Avoid handling of experimental materials and supplies on bench tops, in refrigerators, incubators, freezers, and biological safety cabinets.

V. Standard Operating Procedures

A. BSL-3 Entry Authorization

The procedure to grant access to an individual to the TB BSL-3 lab is (not in a strict order):

1. An individual must first have a background check initiated by HRMS (Human Resource Management Service). In accordance with 1200.06 (Control of Hazardous Agents in VA Research Laboratories) this background check must be initiated for BSL-3 access but does not need to be completed for entry into the BSL-3.

7

2. The individual fulfills the medical surveillance requirements of the VA Long Beach Healthcare System Employee Health Clinic. This usually involves a TB skin test and medical evaluation for N95 respirator usage.

3. Fit testing and training in the use of the N95 half face respirator must be completed.

4. The individual will read this TB BSL-3 SOP which includes the TB Laboratory Safety Manual, Security Plan and SOP.

5. The individual will complete the online Biosecurity training class.

6. The TB lab director makes a formal written request for access by submitting two forms. One is the “Request for Staff Access to Research Secured Laboratory” form. Also submitted will be the “Access to Tuberculosis BSL-3 Lab Checklist”.

7. The individual will complete the “Application for Access to Research Secured Area”.

8. At this point, in accordance with 1200.06 (Control of Hazardous Agents in VA Research Laboratories), the individual will be granted escorted access to the BSL-3 for the purpose of TB specific training. This will be under the supervision of the TB lab director. During this time the TB lab director will ensure that all required PPE is used. The individual will sign into the BSL-3 log book each time which will serve as documentation of training. The correct handling of cultures, how to avoid creating an aerosol, safety and security procedures for the BSL-3 will be covered.

9. The Subcommittee on Research Safety (SRS) will review: a. Access to Tuberculosis BSL-3 Labb. Application for Access to Research Secured Areac. Access to Tuberculosis BSL-3 Checklistd. N95 Respirator fit testing certificate e. Biosecurity training certificate

The SRS will request that an individual be granted full access to the BSL-3. The minutes from the meeting must reflect that all documents were reviewed. (SRS is delegated by the R&D to perform this function

10. The BSL-3 access request is sent to the Medical Center Director for his approval and signature.

11. A copy of the signed request is sent to the Chief of Police so that the BSL-3 access code can be updated. The individual will now have unescorted access to the BSL3.

All signed copies of documents will be filed in the Research Office. Training in the safe and secure handling of Mycobacteria will not stop with the granting of full access to the BSL-3. Training of the individual - as well as the TB lab director - is an ongoing and continuous process.

R&D will review all individuals with access to the TB Lab BSL-3 semi-annually.

B. Protocol Submission

All individuals using the BSL-3 Facility must follow established guidelines. Persons failing to comply will be denied access to the Facility pending a review by the Lab Director.

All research projects must be submitted in writing to the Subcommittee on Research Safety. The project will be reviewed and approved by the Subcommittee on Research Safety. The assessment should include a risk analysis of the organism under investigation and any research methods that may affect that risk (e.g., procedures requiring highly concentrated amounts of bacteria). No human or animal pathogens may be studied without prior approval of the Subcommittee on Research Safety.

8

C. Personnel Training

The training procedure is divided into four stages:

1. Knowledge of the safety manual and protocols2. Training in the use of personal protective equipment and N95 respirators3. Observation of trained personnel working with infectious material4. Working with infectious material while supervised by trained personnel

The Laboratory Director or designee will review general guidelines for working with infectious agents with the trainees, who must show familiarity with the safety manual procedures and be knowledgeable of the potential health hazards of working with the organisms under study. The trainee must also demonstrate proper use and maintenance of room equipment.

Unescorted entry into the BSL-3 Facility is restricted to authorized and approved users (see Section C, Research personnel User). Maintenance or repair personnel, contractors, inspectors and visitor will be allowed only with prior notification and approval of the Lab Director (Section IV).

D. Work Practice Controls

Work practice controls reduce the likelihood of employee exposure to infectious agents by altering the manner in which a task is performed. They provide a margin of safety for unrecognized hazards. Safety is a shared responsibility and the proper attitude and use of safe work practices are critical. The following work practice controls will be enforced within this Facility.

All necessary safety materials and exposure control equipment must be available and in good working order.

A tuberculocidal disinfectant (for example 5% Lysol I.C. or a 1:10 dilution of bleach) and paper towels will be kept nearby in case of a spill.

Mechanical pipetting devices are required. Mouth pipetting is not permitted. All breaks or cuts in the skin must be covered with a bandage. It is left to the discretion

of the Director to prohibit any individual employee from working in the Facility due to an injury that may increase the risk of laboratory acquired infection.

There will be minimal usage of needles or sharp objects within the BSL-3 Facility. If sharps must be used, personnel must employ extreme caution. Needles and sharps are to be disposed of in puncture-resistant, labeled, leak-proof containers. Needles should not be recapped, bent, or cut. If recapping is necessary a one-handed technique must be utilized.

Personnel must keep an uncluttered workspace, especially in the biosafety cabinets. No personal material will be brought into the BSL-3 Facility. No food or drinks is allowed in the BSL-3 Facility. No applying cosmetics in the BSL-3 Facility. The phone in the Facility is intended for emergencies and lab-related needs. It must not

be used for personal business. Always leave respirators in place when using the phone.

E. Entry Preparation

All personnel must follow the following procedures:

1. While in the Ante-Room, the entrant dons necessary PPE which may include:

Personal protective wrap-around or solid front gown N95 Respirator One pair of gloves Eye protection or head covering

9

2. All work to be done in the inner rooms should be defined in advance to ensure that all needed supplies are brought in with the operator at the start of the work, avoiding exit precautions.

The person is now ready to enter the laboratory.

F. Exit Procedures

1. If your gown is soiled, remove and throw it out in the biohazard container. If unsoiled, remove and label it with your name and place it on the hooks provided in the Ante-room. Gowns will be autoclaved before being washed by the VA Laundry service.

2. Remove gloves and place them in the biohazard container.

3. Wash hands with germicidal soap. Remove N95 respirator and place on hooks located on the wall next to the exit door of the Facility. If your mask is damaged, soiled, wet or used for extended periods of time (several hours) it must be discarded.

4. The user may now exit the Ante-room. After exiting the Ante-room, check that the door is closed and locked.

VI. General Lab Procedures The greatest danger in the BSL-3 Facility is the exposure to infectious aerosols. The production of aerosols may result from pouring liquid cultures or supernatants, mixing a liquid culture using high speed devices such as a vortex, dropping a tube or container of liquid culture, or breaking a tube during centrifugation. (See Section C, Working with Organisms).

Disinfection: Mycobacteria can be killed by UV, germicidal solutions such as Lysol I.C., Amphyl, bleach, autoclaving, or heating (>10 minutes) at 80°C.

A. Biological Safety Cabinets

All work with viable infectious organisms must be conducted in the biological safety cabinet. Some procedures generate significant aerosols; they include pipetting, transfer operations, using a blender, centrifugation, sonication, and shaking. Care should be taken to conduct these procedures away from the open face of the cabinet.

1. Before working in the biosafety cabinet, the UV fixture must be off and the fluorescent light on. Place all materials in the cabinet at the beginning of the procedure to limit the number of times that hands pass through the air barrier. Arrange the materials within the cabinet in a manner to ensure aseptic technique; i.e., all work flows from one side of the cabinet (clean) to the other side (dirty). Contaminated items never cross over uncontaminated/clean supplies.

2. Blocking the intake grills in the front and rear of the cabinet interferes with proper functioning of the cabinet. Place large items close to the sidewalls, rather than at the back of the cabinet where they will interfere with airflow.

3. Use a pipette sterilizing can with a small amount of 5% Lysol I.C. covering the bottom for contaminated waste, e.g., pipette tips, transfer pipettes, etc. A 2 liter trap containing ~100 ml stock Lysol I.C. is to be used for liquid non-radioactive waste. All traps connected to the vacuum require filters in the lines. Items must be properly decontaminated by the user following an experiment. All other necessary equipment and supplies for studies involving radioactivity will be the responsibility of the investigator. Each user must follow proper techniques and surveillance of work with radioactivity. (NOTE: Any use of radioactivity in the biosafety cabinets must have prior approval through the Radiation Safety Committee.)

4. Every individual is responsible for cleaning the Laminar Flow hoods when they have finished working. This cleanup involves the following steps: Return all cultures to the incubator or refrigerator, or place in a biohazard bag to be

autoclaved.

10

Remove all items from the biosafety cabinet, except for those listed in item 6 (below). Items used in the cabinet must not be removed from the room without disinfecting.

Wipe down all surfaces in the hood with 5% Lysol I.C., including the walls and working surface.

Spray down the same surfaces with 70% ethanol, removing the sticky film left by the Lysol I.C. solution.

Turn the UV light on. The UV must be on for a minimum of 20 minutes after completing infectious work before the next user may work in the hood.

5. Discard all paper towels in the room biohazard waste container.6. The cabinet must be left decontaminated and ready for the next user. The following items may

remain in the hood and do not need to be cleaned unless they were known to be contaminated or must be removed for purposes of the experiment: Pipette tip box

Pipetteman

Amphyl trap

Pipette Aid

Vortexer

Pasteur pipette container

B. Shipping and Receiving Specimens

All containers received must be opened in a biosafety cabinet. The shipping of specimens and cultures containing, or suspected of containing, live mycobacteria must be carefully controlled to ensure that no live bacilli are released. Disinfect the exterior of the culture or sample tubes before placing into the shipping tubes. The recommended system is to place the tube containing the sample in a screw top container containing an absorbent pad. This container is placed in a cardboard mailer and these in turn are placed in a sealed Tupperware or Styrofoam box. Specimens sent in liquid media shall be in screw top tubes in a sealed plastic storage rack, which is placed in a Tupperware or Styrofoam box lined with an absorbent pad. Specimens packed in this manner may be sent through the mail or delivered by courier service directly to a BSL-3 Facility (See Appendix 2. Packaging and Labeling of Etiologic Agents).

C. Working with Organisms

It is important to remember that tuberculosis is an air-borne pathogen and when working with live bacilli and other organisms in this Facility, the worker must minimize or avoid generating aerosols. Common procedures that could conceivably create aerosols are:

Pouring liquid cultures in open containers

Mixing or stirring liquids in open containers

Using fixed volume automatic pipetting

Rapid pipetting

Opening NRP-2 tubes

Uncapping tubes

Tubes breaking in the centrifuge

Dropping cultures

11

Cultures on slants, plates, or in broth must be properly sealed and the exterior wiped with disinfectant before removing them from the hood. Broth cultures must be kept in disposable, thick-walled culture tubes with screw caps, plastic screw capped tubes, or 0.25 filter caps. If there is any question regarding the acceptability of your containers, have them approved by BSL-3 personnel before use.

D. Specimen Transporting/Removal from Facility

Live cultures brought out of the Facility must be properly disinfected, packaged and identified. Wipe the exterior of any tube with 5% Lysol I.C., followed by 70% ethanol. Cultures must be placed in a double mailer that has the original tube inserted into a plastic bag, surrounded by absorbent material, and placed in a screw cap container. Identification is placed around that container and is inserted into another container that is labeled with a biohazard symbol.

E. Specimen Storage

Samples are to be stored in the refrigerator, freezer, or incubators only. All specimens must be labeled with content, and date. When a sample is no longer needed it must be autoclaved and properly discarded according to LBHCS Infection Control guidelines on waste management. Unlabeled specimens will be discarded immediately. Samples may be stored in the incubator for up to two months and in the refrigerator for three months; after this time they will be discarded. Individuals are responsible for maintaining and disposing of their own samples.

All samples stored in the refrigerator or the incubators must be checked at least once a week for signs of contamination. Samples (both agar and broth cultures) growing mold must be discarded as soon as possible to avoid contaminating other cultures and equipment. The high humidity level is conducive to mold spread into the entire incubator chamber, placing everyone's work at risk.

Samples found to have gross contamination must be removed. Notify BSL-3 personnel so the proper individual can also be notified of the incident. Following notification of the contaminated samples, the individual will be given a specified time in which to remove contaminated samples.

All specimens stored in the freezer must be placed inside labeled and covered storage boxes. For specimen freezer storage use an appropriate tube. All samples must be marked properly or they will be discarded.

F. Biohazard Waste

All slants, plates, and cultures no longer needed must be autoclaved. Place waste in the biohazard waste container by the autoclave. When about three-fourths full, the bags should be tied shut, and an indicator tape placed on the outside of the bag. The bag is then autoclaved and disposed of in the larger biohazard trash can in the Ante-room.

All infectious waste generated in the biosafety hood must be placed in the receptacles containing about one to two centimeters of 5% Lysol I.C., or 1:10 bleach. Containers of 5% Lysol I.C. and 1:10 bleach are located near each biocabinet. When work is completed, remove the covered receptacle from inside the biocabinet and autoclave.

Sharps containers are to be used for all needles and other sharp items, e.g., glass slides, scalpels, blades, and pasteur pipettes.

Liquid wastes should be placed in an appropriate container for autoclaving. Contaminated liquid waste must never be poured down the sanitary drains. If a contaminated liquid is spattered within the hood or glove box, the affected area should be covered with a disinfectant-soaked towel for 5 to 10 minutes, then the towel(s) used must be bagged with the contaminated trash and autoclaved as soon as possible.

12

G. Autoclaving Waste in the Facility

All infectious waste (dry and liquid) generated must be autoclaved before removal from the Facility.

1. Dry Waste

Dry waste items are to be placed into the biohazard bags after their use. The bag is then tied or taped shut (making sure to leave some empty space in the bag for expansion during autoclaving), sterilization tape placed on and then autoclaved on a dry cycle.

2. Liquid Waste

Liquid waste may not be disposed of down the sink before it has been autoclaved. It is to be placed into autoclavable containers, sterilization tape placed on the outside, and autoclaved. After sterilization, the liquid may then be disposed of down the sink.

H. Radiation Use

The BSL-3 Facility is under a sub-license from the Radiation Safety Committee and any individuals not following the rules will not be allowed to work in the Facility.

1. All users of the Facility must adhere to the following:

Follow proper rules for working safely with radiation and disposal of radioactive waste

Wear a radiation badge under their gowns when working with radiation if required

Use only equipment (centrifuge, incubator, pipette, etc.) labeled for work with radiation

2. BSL-3 Facility personnel are responsible for the following safety activities:

Conduct monthly swipes and counter readings (using the appropriate counter) of radioactive work areas to check for contamination. Any contaminated areas will be cleaned up and checked again. If there is no recorded use of radiation in the logbook for that month, this may be noted on the radiation record sheets and no swipes or counter readings are required for that month. If any contaminated areas are detected, BSL-3 Facility personnel will contact the individual(s) responsible for the spill so that they can decontaminate the area.

Maintain records of monthly swipes and counter readings in a binder in room 203. Include indication that decontamination of contaminated areas has been accomplished.

I. Facility Maintenance

All users of the Facility must perform the following responsibilities:

Check both the jacket water supply level and the tray in the bottom of each chamber in the incubators.

Check the readouts on the biosafety cabinets, freezer, and incubators. Notify BSL-3 personnel if equipment is not operating properly.

Keep equipment clean and notify BSL-3 personnel of maintenance needs.

Clean and generally maintain the laboratory. Keep all necessary supplies in the antechamber. These include a dust broom, spongemop, bucket, bleach, 70% ethanol, and paper towels.

Mop the floor using bleach as necessary. Counter tops and equipment must be wiped down with 70% ethanol once a month.

Wipe down the UV lights in the biocabinets with ethanol once a month. Prior to cleaning, they should be turned off and allowed to cool for ten minutes. Fluorescent light bulbs will be changed when they go out. UV lights will be replaced when the UV levels drop below germicidal limits as monitored by a UV meter.

13

Check the CO2 levels of the gas tanks.

Replace the data-log roll on the autoclave when empty.

VII. Emergency Procedures

A. The BSL-3 Facility Air-Flow System

Proper airflow must be maintained at all times. An airflow system is an integral part of the design of the Facility, and is monitored by AC personnel, who are available 24 hours/day. VA maintenance personnel will ensure correct operation of air balance system every year.

Operating FailuresThe air handling and exhaust systems are interlocked. In the event the exhaust fails, the supply air shuts down. If the supply air malfunctions, the exhaust must remain on. The indicator panel in the anteroom indicates if there has been a failure of the ventilation system.

There are two possible failures. (1) The biosafety cabinets, but not the room air exhaust system, is on emergency power. If there is a power failure in the research building the air exhaust will shut down but the biosafety cabinets will continue to maintain negative pressure. If this occurs all work in the room will stop until the power returns. (2) If both the room air exhaust system and biosafety cabinet fail this will results in the complete loss of negative pressure in the BSL-3. If this happens all work in the BSL-3 will be stopped immediately. All personnel will leave and the room will be seal shut with duct tape until the room is repaired.

B. Spills

Accidental spills of mycobacterium cultures or clinical specimens require special considerations in regard to decontamination procedures. All spills must be immediately reported to BSL-3 Facility personnel. Spills are divided into three types and should be dealt with accordingly.

1. Spills within biosafety cabinets are handled immediately by the worker. Leave the biosafety cabinet running and cover the spill with a generous amount of 5% Lysol I.C. or 1:10 bleach. After 15 minutes, wipe up the liquid with paper towels. Equipment or supplies affected by the spill should be sprayed with 5% Lysol I.C. or 1:10 bleach and then wiped dry. Surfaces are then sprayed with 70% ethanol. All towels are disposed of in the biohazard waste container in the room.

2. Accidents outside the biosafety cabinets, such as spilling a culture or breaking a rack of slants, require the following action:

Everyone in the room immediately exits.

In the anteroom of the Facility, remove gown, and gloves and dispose of them in the biohazard waste. Wash hands with germicidal soap and water and then remove the disposable respirator.

Everyone leaves through the outer door. A sign will be placed on the outer door to the Facility forbidding entrance.

Contact BSL-3 personnel. See Appendix 4 for Emergency Call list in the event of a spill.

After one hour, the person responsible for the spill and BSL-3 personnel will enter the Ante-room. Employees must enter wearing standard BSL-3 PPE.

BSL-3 personnel will direct the clean up. After entering the inner room, the spill is covered with the powdered absorbent found in the spill kit, located in the Ante-room. Care must be taken to gently apply the powder in order not to re-aerosolize the spill. The absorbed infectious spill is then gathered and scooped into the biohazard container.

14

The contaminated area on the floor is sprayed with 5% Lysol I.C. and wiped dry with a paper towel. All affected equipment, counters, and the walls must be sprayed with the disinfectant, allowing contact of at least 20 minutes, and then wiped dry. The floor must be mopped with a 1:10 bleach solution.

3. Major spills, such as a large culture that drops and splatters infectious broth throughout the room, a full shelf of the incubator toppling to the floor and sending slants and broth in all directions, etc., or a release occurring during centrifugation, must be dealt with in the following manner:

Everyone in the room must immediately stop what they are doing, properly dispose of their outer pair of gloves, and leave the room.

In the Ante-room of the Facility, remove gown, and gloves and dispose of them in the biohazard waste. Wash hands with germicidal soap and water and remove the disposable respirator.

Seal the inner door with duct tape. Exit through the outer door. Contact BSL-3 personnel. See Appendix 4 for the Emergency Call List in the event of a

spill. BSL-3 personnel, equipped with a RACAL PAPR will assess the degree of contamination

and determine if the Facility will need to go through a decontamination procedure. Spills that occur in equipment such as a centrifuge must be sprayed with 5% Lysol I.C. (not

bleach), allowing 20 minutes (minimum) contact time, then wiped dry.

C. Exposure Incidents

If a person working in the Facility spills contaminated material on exposed skin, the affected area should be washed with germicidal soap. If contaminated material is spilled on clothing, the affected article of clothing must be removed and placed in a biohazard bag, which is taped shut and must be autoclaved.

If an employee has reason to believe he/she has been exposed to Mycobacterium tuberculosis or any other biological or chemical material through contact, inhalation, or puncture, the following steps must be followed:

Report the incident to the Director (or designee) who will in turn notify appropriate department personnel.

Notify anyone else in the area that may also have been exposed. An Employee Accident Injury Report will need to be completed. These are available in the

VAHS Research Office. The VA Occupational Health Clinic should be consulted and a PPD skin test or other tests, as

deemed necessary, scheduled.

D. Power Failure

In the event of a black out, the employee's actions are determined by the response of the building's back-up generator. The loss of power to the building activates the emergency generator and power and should be restored within 50 seconds. If the power and lights are not restored after one minute, the operator must stop work and leave the room. Emergency lights in the corridor will activate to provide exiting capabilities. If the power remains off, the door to the inner chamber should be sealed with duct tape. Safety apparel is then properly discarded, and individuals may leave the Facility, after proper hand washing.

E. Fire

1. Evacuation

15

If the fire can be contained, extinguish the fire using a fire extinguisher. The Ante-room has an extinguisher available. If not able to contain the fire, evacuate immediately, and notify emergency personnel. Report to the fire fighting authorities the specific agents, animals and special hazards associated with the BSL-3 Facility.

2. Cleanup and Disinfection

After a fire affecting the BSL-3 Facility, personnel gowned, gloved, and using a RACAL PAPR must assess the situation to determine the extent of contamination in the Facility and whether disinfection of the surfaces or a full decontamination of the Facility is required before resuming activities.

F. Earthquake

The strength of an earthquake can vary and the employee’s actions are determined by the strength of the earthquake and resulting damage. The first step after a quake is to determine if the ventilation of the BSL-3 is still intact. This can be done by consulting either of the 2 air pressure monitors in the lab. If the ventilation is not functional then the door to the inner chamber should be sealed with duct tape. Safety apparel is then properly discarded, and individuals may leave the Facility, after proper hand washing. All damage is reported to the Engineering Section

16

Table

Inner Room9'7"x9'10"

808 ft2

Hood 2 Hood 1

Bench

Sink

Table

Autoclave

Shaker 2-37°CIncubators

Centrifuge

Centrifuge

37°CIncubator

-80°CFreezer

Fridge

Outer Room9'x20'6"1568 ft.2

CO2 Tank

Bench

Sink

Bench

Shelf

Appendix 1

Map

17

Appendix 2

Packaging and Labeling of Etiologic Agents

Safe Transport of Infectious Substances 1)

IntroductionThese guidelines are applicable to the transport of infectious substances both nationally and internationally. They provide information for identifying the material to be transported and for its safe packaging and transport. The guidelines stress the importance of developing a working relationship between the groups involved – the sender, the carrier and the receiver – in order to provide for the safe and expeditious transport of this material.

The packaging of infectious materials for transport must therefore address these concerns and be designed to minimize the potential for damage during transport. In addition, the packaging will serve to ensure the integrity of the materials and timely processing of specimens. The international regulations for the transport of infectious materials by any mode of transport are based upon the Recommendations of the United Nations Committee of Experts on the Transport of Dangerous Goods (UN). The Universal Postal Union (UPU) reflects these recommendations in its regulations, particularly for packaging. The International Civil Aviation Organization (ICAO) and the International Air Transport Association (IATA) have also incorporated the UN Recommendations in their respective regulations, as have other international transport organizations. The World Health Organization serves in an advisory capacity to these bodies. This document provides practical guidance to facilitate compliance with current international regulations. If, at a future date, any modification is made in the section of the UN Recommendations on the Transport of Dangerous Goods dealing with infectious substances and diagnostic specimens, these guidelines will be updated accordingly.

DefinitionsFor the purpose of describing transport safety measures the terms “infectious substances” and “infectious materials” are considered synonymous. The term “infectious substances” will be used in this document.

Infectious SubstancesAn infectious substance is defined as a substance containing a viable microorganism, such as a bacterium, virus, rickettsia, parasite or fungus, that is known or reasonably believed to cause disease in humans or animals.*

With respect to packaging and transport situations, infectious substances include:1. All cultures containing or suspected of containing an agent which may cause infection2. Human or animal samples that contain such an agent in quantities sufficient to cause infection, should an

exposure to them occur due to a transport mishap3. Sample(s) from a patient with a serious disease of unknown cause4. Other specimens not included above and designated as infectious by a qualified person, e.g., a physician,

scientist, nurse, etc.* This definition is taken from the current UN Recommendations on the Transport of Dangerous Goods. Prions are not included in this definition although they are considered to be infectious agents.

Packaging, Labeling, and Documentation for TransportThe packaging requirements are determined by the UN and are contained in ICAO and IATA regulations in the form of Packaging Instructions (PI) 602.

Basic Triple Packaging SystemSee Figure A2-1

The system consists of three layers as follows:1. Primary receptacle. A labeled primary watertight, leak-proof receptacle containing the specimen. The

receptacle is wrapped in enough absorbent material to absorb all fluid in case of breakage.

18

2. Secondary receptacle. A second durable, watertight, leak-proof receptacle to enclose and protect the primary receptacle(s). Several wrapped primary receptacles may be placed in one secondary receptacle. Sufficient additional absorbent material must be used to cushion multiple primary receptacles.

3. Outer shipping package. The secondary receptacle is placed in an outer shipping package that protects it and its contents from outside influences such as physical damage and water while in transit. Specimen data forms, letters and other types of information that identify or describe the specimen and also identify the shipper and receiver should be taped to the outside of the secondary receptacle.

Figure A2-1Requirements for Infectious SubstancesThe basic triple packaging system is used with the following additional specifications, labeling, and documentation requirements. Infectious substances may only be transported in packaging which meets the UN class 6.2 specifications and packaging instruction (PI) 602. The outer shipping package must bear the UN Packaging Specification Marking (Figure A2-2).

Figure A2-2

Hand carriage of infectious substances is strictly prohibited. The maximum net quantity of infectious substances which can be contained in an outer shipping package is 50 ml or 50 g if transport is by passenger aircraft. Otherwise, the limit per package is 4L-4Kg for transport by cargo aircraft or other carriers. Primary receptacles exceeding 50 ml in combination packing must be oriented so the closures are upward, and labels (arrows) indicating the “UP” direction must be placed on two opposite sides of the package. The passenger aircraft quantity limits do not apply to blood or blood products for which there is no reason to believe they contain infectious

19

substances, when in receptacles of not more than 500 ml each and with a total volume of not more than 4L in the outer package.

Hazard labels for dangerous goodsFor all dangerous goods to be shipped by airfreight, specific hazard label(s) must be affixed to the outside of each package. The following hazard labels are of importance for culture collections or other institutions shipping biological substances.

Hazard labels for infectious substances and for genetically modified microorganisms that meet the IATA definition of an infectious substance:

Figure A2-3

Name: Infectious Substance Minimum dimensions: 100 x 100 mmFor small packages: 50 x 50 mm (black and white)

Hazard label for noninfectious genetically modified microorganisms and for carbon dioxide, solid (dry ice):

Figure A2-4

Name: MiscellaneousMinimum dimensions: 100 x 100 mmFor small packages: 50 x 50 mm (black and white)Hazard label for liquid nitrogen:Name: Non-flammable gasMinimum dimensions: 100 x 100 mmFor small packages: 50 x 50 mm (green and white)

20

Packages containing liquid cultures of infectious organisms and genetically modified microorganisms must be packed so that the closure(s) of the inner packaging(s) are upward; the upright position of the packaging must be indicated by two “Package Orientation” labels (black or red arrows). The labels must be affixed on opposite sides of the packaging. A label “THIS SIDE UP” or “THIS END UP” may also be displayed on the top cover of the package:

Name: Package Orientation (see label on outer container of Figure A2-1)Minimum dimensions: 74 x 105 mm (black or red and white)For small packages of infectious substances dimensions may be halved.

In case shipments include only freeze-dried cultures the quantity should be given in g or mg. not in ml. The “PACKAGE ORIENTATION” labels should be affixed to avoid any delay.

Labeling of the outer package for shipment of infectious substances must include the elements listed hereafter.

1. The International Infectious Substance Label.

2. An address label with the following information: The receiver’s (consignee) name, address and telephone number The shipper’s (consignor) name, address and telephone number The UN shipping name (infectious substances affecting humans or animals as the case may be) followed by The scientific name of the substance The UN number (humans – UN2814, animals UN2900) Temperature storage requirements (optional)

If the outer package is further packed in an overpack (with dry ice for instance) both outerpack and overpack must carry the above information, and the overpack must have a label stating “INNER PACKAGES COMPLY WITH PRESCRIBED SPECIFICATIONS”.

3. Required shipping documents – these are obtained from the carrier and are fixed to the outer package: The shipper’s Declaration of Dangerous Goods (Figure 5 is one example) A packing list/proforma invoice which includes the receiver’s address, the number of packages, detail of

contents, weight, value (NOTE: state that there is “no commercial value” as the items are supplied free of charge)

An airway bill if shipping by air.

4. An import and/or export permit and/or declaration if required.

5. If the outer package contains primary receptacles exceeding 50 ml in combination at least two “Orientation Labels” (arrows) must be placed on opposite sides of the package showing correct orientation of the package.

Requirements for Air MailInfectious substances may be shipped by registered airmail. The basic triple packaging system is used with the same requirements as for other means of conveyance.

The address label must display the word “LETTER” and the green Customs Declaration Label for Postal Mail is required for international mailing. Infectious substances are to be identified with the International Infectious Substance label (see Figure A3-3). Infectious substances must also be accompanied with a shipper’s Declaration of Dangerous Goods form (see Figure A3-5 at the end of the document). Because of local/international restrictions, prior contact should be made with the local post office to ascertain whether the packaged material will be accepted by the postal service.

RefrigerantsIce or dry ice when used in a shipment must be placed outside the secondary receptacle. If wet ice is used it should be in a leak-proof container and the outer package must also be leak-proof. The secondary receptacle must be secured within the outer package to prevent damage after the refrigerant has melted or dissipated. Dry ice must

21

not be placed inside the primary or secondary receptacle because of the risk of explosions. An overpack (a specially designed insulated outer package) may be used to contain dry ice. The outer package must permit the release of carbon dioxide gas if dry ice is used. UN Packing Instruction 904 must be observed. If dry ice is used for infectious substances, the details must appear on the shipper’s Declaration for Dangerous Goods. In particular, the outer most packing must carry the “MISCELLANEOUS” hazard label for dry ice (see Figure A2-4). If liquid nitrogen is used as a refrigerant, special arrangements must be made in advance with the carrier. Primary receptacles must be capable of withstanding extremely low temperatures and appropriate packaging requirement of the carrier must be observed. In particular, the outer most packing must carry the “NON-FLAMMABLE GAS” label for liquid nitrogen.

Local Surface TransportThe principle of safe local surface transport is the same as for air or international transport – the material should not have any possibility of escaping from the package under normal conditions of transport. The following practices should be observed:

Specimen containers should be watertight and leak-proof If the specimen container is a tube, it must be tightly capped and placed in a rack to maintain it in an upright

position Specimen containers and racks should be placed in robust, leak-proof plastic or metal transport boxes with

secure, tight fitting covers The transport box should be secured in the transport vehicle Each transport box should be labeled appropriately consistent with its contents Specimen data forms and identification data should accompany each transport box A spill kit containing absorbent material, a chlorine disinfectant, a leak-proof waste disposal container, and

heavy duty reusable gloves should be kept in the transport vehicle

Transport PlanningIt is the responsibility of the sender to ensure the correct designation, packaging, labeling and documentation of all infectious substances and diagnostic specimens. The efficient transport and transfer of infectious materials requires good coordination between the sender, the carrier and the receiver (receiving laboratory), to ensure that the material is transported safely and arrives on time and in good condition. Such coordination depends upon well-established communication and a partner relationship between the three parties. All have specific responsibilities to carry out in the transport effort.

The Sender: Makes advance arrangements with the receiver of the specimens including investigating the need for an

import permit Makes advance arrangements with the carrier to ensure:

that the shipment will be accepted for appropriate transport that the shipment (direct transport if possible) is undertaken by the most direct routing, avoiding arrival at

weekends Prepares necessary documentation including permits, dispatch, and shipping documents Notifies the receiver of transportation arrangements once these have been made, well in advance of expected

arrival time

The Carrier: Provides the sender with the necessary shipping documents and instructions for their completion Provides advice to the sender about correct packaging Assists the sender in arranging the most direct routing and then confirms the routing Maintains and archives the documentation for shipment and transport Monitors required holding conditions of the shipment while in transit Notifies the sender of any anticipated (or actual) delays in transit

The Receiver: Obtains the necessary authorization(s) from national authorities for the importation of the material

22

Provides the sender with the required import permit(s), letter(s) of authorization, or other document(s) required by the national authorities

Arranges for the most timely and efficient collection on arrival Immediately acknowledges receipt to the sender

Shipments should not be dispatched until: Advance arrangements have been made between the sender, carrier, and receiver The receiver has confirmed with the national authorities that the material may be legally imported The receiver has confirmed that there will be no delay incurred in the, delivery of the package to its

destination.

1) Adapted from "Guidelines for the Safe Transport of Infectious Substances and Diagnostic Specimens," World Health Organization, 1997.

23

Figure A2-5

24

Appendix 3

Respirator Program

The Tuberculosis Control Program is contained in the VA Healthcare System Infection Control Manual. All personnel must wear an air purifying respirator with a rating of N95 or higher in the BSL3 Facility. A PAPR will be used for emergency situations (spill cleanup).

N95 Respirator Use

1. The N95 respirators will be available directly outside the BSL-3 Facility area with other PPE.2. Each N95 respirator must be worn by only one individual.3. The N95 respirator should be inspected before use to ensure the integrity of the components. 4. Each employee requiring the use of a N95 respirator must be fit-tested and complete a medical

evaluation form. 5. The staff will perform a fit check prior to entering the BSL-3 Facility.

Training

All staff members who are required to wear an N95 respirator must receive training that includes the following:

1. The hazards associated with airborne Mycobacterium tuberculosis and the consequence of infection.2. The measures that have been taken to prevent exposure to airborne M. tuberculosis.3. The proper use and limitations of the respirator.4. The proper way to wear the respirator.5. Methods for donning the respirator and checking the face seal. 6. Method for performing the qualitative fit-test procedure.7. A demonstration on how to wear and adjust the respirator.

Fit-testing

1. All staff required to wear an N95 respirator must be fit-tested initially and annually thereafter. Procedure for fit testing is contained in the Tuberculosis Control Program.

2. Staff members may not waive their responsibility to wear proper respiratory protection. Beards or other facial hair at the area of the respirator seal prevent a proper fit test and therefore, these staff members may not use the N95 respirator. For bearded employees in whom a PAPR would not interfere with the performance of their duties, a PAPR will be made available.

3. All fit-testing must be documented. Records will be kept by the individual and copies by the lab director..

Powered Air Purifying Respirators (PAPR)

1. An Air-Mate HEPA 12 PAPR will be available for emergency situations (small spill, etc.) and for bearded employees. These are located outside the Ante-room, in the Dressing Area (see Facility diagram, Appendix 1).

2. Training on the use of PAPRs will be conducted through the Safety Office.

25

AIR-MATE HEPA 12 Powered-Air Purifying Respirator (PAPR)

Principle of OperationThe Air-Mate HEPA 12 head cover is a lightweight headpiece made of polyethylene coated tyvek. It is available in regular and large sizes. The head cover has an absorbent sweatband and is available in regular and large sizes.

Ambient air is drawn through the belt-mounted HEPA Air Filter Unit and passes through the Breathing Tube connected to the Head Cover. The filtered air then passes over and in front of the user's head and face, providing filtered, respirable air. The face seal maintains the air within the breathing zone. Air is expelled at the edge and through the bottom holes of the face seal.

Pre-operational InspectionPrior to using the Air-Mate HEPA system, examine the head cover for physical damage. If any part of the head cover is damaged or torn, discard it. Inspect the breathing tube and the body of the HEPA filter unit for damage. Replace all damaged parts before proceeding.

Ensure that the battery pack is fully charged. Check the airflow as follows:1. To connect the breathing tube to the HEPA air filter unit, insert the breathing tube (male end with

pin) into the HEPA air filter unit and twist clockwise to lock in place. Hold the free end of the tube up by grasping the slotted connector and covering the slots of the connector with thumb and forefinger. Drop the black, bullet-shaped Airflow Indicator (pointed end first) into the slotted connector.

2. Switch the HEPA Air Filter unit ON. Hold the tube so that it is vertical and at eye level. The indicator should 'float' on the air coming out and the lower band should be above the connector's rim.

3. If the lower band on the indicator rises above the slotted connector edge, airflow is sufficient. If the indicator fails to rise to this level, check the breathing tube, battery, and filter.

After assuring yourself of sufficient airflow, remove the Airflow Indicator and connect the breathing tube to the headpiece, by inserting the free end of the breathing tube into the rear of the headpiece until it snaps in place.

System Assembly and DonningThe Air-Mate HEPA system should only be assembled in a non-contaminated environment. Before donning the system, perform the pre-operational inspection on the HEPA air filter unit, head cover, and breathing tube. Connect the breathing tube to the head cover as described above. To properly don the system, place the HEPA air filter unit (with the breathing tube on top) against your lower back along your spinal column. Holding it in place, fasten the belt buckle and ensure the HEPA air filter unit rests comfortably on your lower back. Turn the HEPA air filter unit ON by pushing the black switch on the HEPA air filter unit.

Put the head cover on. Make sure that it fits properly by checking the following:1. The sweatband and elastic encircle your head, with the sweatband against your forehead2. The face shield is pulled as far as possible under your chin and is hugging your face3. The head cover sits just above your ears4. The head cover does not shift on your head (f the head cover does shift on your head, try another size

head cover)5. Check that air flows to the front of the head cover and ensure that the breathing tube is firmly

secured to the HEPA air filter unit and head cover, and is not twisted after the system has been donned.

WARNING: No protection against particulates is provided unless the head cover is correctly positioned over your head, the visor and face seal are correctly positioned in front of your face, and an adequate

26

supply of filtered air is maintained. If the airflow ceases, reduces, or suddenly increases, leave the work area immediately and check the head cover, breathing tube, and HEPA air filter unit for faults.

Post-operational MaintenanceNOTE: For your protection, the Air Mate HEPA System must only be removed in a non-contaminated environment.

When you are ready to remove the respirator, enter a non-hazardous environment and remove the Air-Mate HEPA system in a safe manner.

Disassemble the breathing tube from the headpiece by pulling apart at the snap connection. Twist the end of the breathing tube (attached to the HEPA air filter unit) counterclockwise to separate the breathing tube from the HEPA air filter unit. Wipe the outside only of the breathing tube with a soft cloth, dampened with 70% ethanol to remove possible contamination.

Air-Mate HEPA Filter Unit

The belt-mounted Air Mate HEPA Filter Unit consists of a motor-driven fan, a HEPA filter, and a rechargeable nickel cadmium battery pack. The motor-driven fan draws contaminated air through the HEPA filter and feeds the purified air via the breathing tube into the rear of the headpiece.

A completely exhausted battery pack should be charged for 14-16 hours. A longer charging period may be used with a smart charger. Charge the battery pack as follows:

1. Insert the AC adapter/charger three-prong pug into the socket on the side of the HEPA air filter unit 2. Plug the AC adapter/charger into a 120-volt AV 60Hz power outlet3. When charging is complete, unplug the adapter/charger from the power outlet before removing the

adapter/charger pug from the HEPA air filter unit

Trouble-Shooting, Filter Change, and Battery Pack Removal and Replacement

Refer to the RACAL Air-Mate HEPA System Instruction Manual for trouble-shooting, filter change, and battery pack information.

Operational Approvals and Limitations

The Air-Mate HEPA Powered Air-Purifying Respirator is NIOSH/MSHA approved (Air-Mate HEPA 10 system approval number =TC-21C-635). The Air-Mate HEPA System can be used only under the following conditions:

1. As respiratory protection against dust, fumes and mists having a time-weighted average of less that 0.05 milligrams per cubic meter; asbestos-containing dusts and mists; radionuclides; and radon daughters attached to dusts, fumes and mists

2. In atmospheres NOT immediately Dangerous to Life or Health (IDLH)3. In ambient air atmospheres where oxygen concentrations equals or exceeds 19.5%4. In working atmospheres that do NOT contain toxic gases or vapors5. When the user has been trained and instructed in the proper use and maintenance of the equipment6. When only RACAL replacement parts have been used to repair the equipment

NOTE: These systems have NOT been tested and are not approved for intrinsic safety.

27

Appendix 4

Emergency Distribution List

For use in an emergency situation of any kind (spill, accidental exposure, mechanical failure resulting in compromise of negative pressure, etc.):*

1. Charles SohaskeyTB Lab Research DirectorLab: (562) 826-8000 ex 4914Home: (562) 533-7207

2. Julie Murphy, RN, Infection ControlOffice: 5481

3. Jim Bachman, SafetyOffice: (562) 826-5464Mobile: (562) 577-6110

4. Facility Officer on Duty (FOD)Phone 3713 or 3714

5. Police and SecurityPhone: 5800

6. Administrative Officer

Phone: 5801

7. Air Handling/Circulation:Normal hours: 3713After hours: 3713/3714

*A NOTE ON EMERGENCIES: In general, remember that the goal in every case is to minimize a release of pathogens from the BSL-3 lab.

In the event a spill occurs, clean-up must be immediate, thorough, and not compromise the safety of other people in the suite or in the building. When entering the BSL-3 to assist spill clean up, you must wear a RACAL powered air purifying respirator and all appropriate protective clothing.

If the emergency involves a mechanical failure, such as shutdown of the negative pressure airflow, the lab is designed with ducts that are held open by the airflow. This will not stop potentially contaminated air from leaking underneath the doors (or through them) when they are opened. If negative pressure is lost, do not open the door, except to leave the lab. Use duct tape to seal the door and put a sign on the door to prevent anyone from entering. If HEPA filtration is lost, but airflow is not compromised, do not turn off the air circulation. Notify the Facility Safety Officer that unfiltered air is being vented and prevent maintenance workers from being exposed. While we are not legally required to HEPA-filter roof-exhausted air, it is tempting to turn off air circulation, to try to prevent unfiltered air from being vented. However, we must first protect against contaminated air leaking out of the lab into the closed environment of the building; at the same time, if air circulation is lost, temperatures within the BSL-3 will climb dramatically.

28

Appendix 5

INCIDENT REPORT FORM

1. List the name(s) of the employee(s) involved:

2. List the date, time and location in which the accidental exposure occurred:

3. As thoroughly as possible, describe the circumstances of the exposure incident:

4. List the pathogens and route(s) of possible exposure (e.g. inhalation, subcutaneous, etc.):

5. What is the nature of the organism strain to which the employee has been exposed? (strain name and history, complete drug-resistance/susceptibility profile, any other information that might be pertinent to treatment):

6. List steps taken to evaluate employee health, and action taken to prevent recurrence of a similar incident:

29

ATTACHMENT A

09-151 Security PolicyVA Long Beach Healthcare System

REQUEST FOR STAFF ACCESSRESEARCH SECURED AREA

1. PURPOSE: To formally request access for Investigator’s employees and staff to the Research Secured Area.

2. POLICY: The information requested in this document must be supplied via submission of this form or Email before access to the secured area will be considered.

3. RESPONSIBILITY: It is the responsibility of each Investigator to formally identify the staff that must have access to the secured area in order to complete their research-related duties.

4. PROCEDURE: The Investigator submits the information requested via this document to the Research Administration. NOTE: Attachment B is also required.

5. REQUESTED INFORMATION: a. Person making request: ________________________________________________

b. Name of person for whom access is requested:_______________________________

c. Immediate supervisor of person for whom access is requested: __________________

d. Brief description of duties of person for whom access is requested (lab support,research assistant,

etc):___________________________________________________

e. Areas to which access is needed. Check all that apply:(1) General Laboratory Area

(2) BSL3 (do not mark this box unless otherwise instructed)

(3) VMU (only mark this box if you expect to work with animal subjects)

____________________________________________Signature of Principal Investigator(This PI must be the investigator assigned the laboratory space where the new staff member will work.)

30

ATTACHMENT B

09-151 Security PolicyVA Long Beach Healthcare System

APPLICATION FOR ACCESS TO RESEARCH SECURED AREA

1. PURPOSE: To formally apply for access to Research Secured Area.

2. POLICY: In order to be considered for access to the secured laboratory area, the applicantmust submit this document to the Research Administration.Note: the Principal Investigator must also complete ATTACHMENT A.

3. REQUESTED INFORMATION:a. Full Legal Name ___________________________________________________________

b. Home Address (not Post Office Box) ___________________________________________

c. Date of Birth __________________

d. Place of Birth ____________________________________________

e. Citizenship Status:(1) U.S. Citizen(2) Other (Attach copy of document indicating legal authority to be in U.S.)

f. Circle “yes” or “no” for each item below. Any “yes” answers should be detailed in 3f(8)below.

(1) Yes No I am under indictment for a crime punishable by imprisonment exceeding 1year.

(2) Yes No I have been convicted of a crime punishable by imprisonment exceeding 1 year.

(3) Yes No I am in fugitive status from any local, state, national, or international lawenforcement agency.

(4) Yes No I am an unlawful user of any controlled substance.(5) Yes No I am an alien illegally or unlawfully in the United States.

(6) Yes No I have been adjudicated as mental defective or have been committed to amental institution.

(7) Yes No I have been discharged from the United States Armed Services underdishonorable conditions.

(8) Comments/Details/Notes:____________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________

______________________________________ ___________________________

31

Signature of Applicant Date

References

Richmond, J. Y., Knudsen, R. C., and Good, R.C. Biosafety in the Clinical Mycobacteriology Laboratory 1996. Clinics in Laboratory Medicine (Ed Heifits, L.B.) 16: 527-550.

NIH Guidelines for Research Involving Recombinant DNA Guidelines (NIH Guidelines). FR34496

Guidelines for Preventing the Transmission of Mycobacterium tuberculosis in Health-Care Facilities, 1994. Morbidity and Mortality Weekly Report 43:RR-13 1-132.

Occupational Exposure to Bloodborne Pathogens; Proposed Rule and Notice of Hearing. 1989. Department of Labor. 29 CFR Part 1910.

BSL-3 Safety Manual. 2000 Second Edition. University of Iowa.

Guidelines for the Safe Transport of Infectious Substances and Diagnostic Specimens. 1997 World Health Organization.

Biosafety in Microbiological and Biomedical Laboratories. 1999. Fourth Edition. CDC and NIH.

32