Attachment_8_SEPRL_Building_45_Biosafety_Manual_APRIL_2021.pdf

SEPRL Biosafety / Biosecurity Guidelines

SEPRL BUILDING 45

LABORATORY BIOSAFETY

MANUAL

BIOSAFETY / BIOSECURITY

GUIDELINES

April 2021 New

Southeast Poultry Research Laboratory

934 College Station Road

Athens, GA 30605

Contents Introduction

Facility SHEM Policy Statement

Security Related Policies

Principles of Biosafety and Biosecurity

Definitions

Facility Operating Levels

Clothing Requirements and Movement of Personnel

General Information

General Clothing Requirements

Laundering Lab-supplied Clothing

Clean Clothing

Specific Clothing Requirements

Buildings 13, 15, 16, 17, 43, 44

Building 45

Biosafety Level 2 Areas

Laboratories

Animal Housing - Buildings 6, 8, and 39

Autoclaves

Autoclaving Sharps

Biosafety Cabinets

Work Practices for Biological Safety Cabinets

Working in a BSC

Centrifuges

Movement of Materials

Working with Biological Agents

Biosafety Level 2

Working with Animals

Animal Biosafety

Entering an Animal Production Building

Buildings 13, 15, 16, 17, 43 or 44

Exiting an ABSL-2E Animal Room

Buildings 6, 8, and 39

Emergency Exit

Use of Controlled Drugs in Animal Experiments

General Cleanup Protocols

Spills - Biohazardous Materials

General Considerations

General Procedure

Procedures for Specific Locations

HEPA Filters – Use, Removal and Disposal

Special Operational Procedures

Transfer of Birds between Buildings within the Compound

SOP: #06 Campus animal transport

Packaging of Biological Materials for Shipment

Shipping and Receiving of Biological Materials

Shipping Biological Materials

Receiving Biological Materials

Inventory and Control of Biological Materials

Definitions:

Experimental samples

High consequence pathogens (HCP)

Low consequence pathogens (LCP)

Repository

SEPRL Biosecurity Advisory Committee

SEPRL Biosafety/Biosecurity Officer/Biosafety Specialist

Working stocks

General Information

Uniform Experiment Numbering

Infectious Materials Classification

High Consequence Pathogens

Low Consequence Pathogens

Appendix

SEPRL LABORATORY EQUIPMENT DECONTAMINATION FORM

Annual Review Verification

Date Signature

SEPRL CONTACT INFORMATION

Dr. David Swayne, SEPRL Lab Director/Institutional Official

706-546-3433, 706-612-6058 (government cellphone), 706-207-4961 (personal cell)

Dr. Anne Hurley-Bacon, USNPRC

Attending Veterinarian

706-224-0926 (cell)

Nick Chaplinski, Biosafety Officer 706-546-3407, 706-614-4275 (cell)

Jeffrey Martin, Supervisor Biological Laboratory Specialist/Operations

706-546-3721, 706-340-0406 (cell)

Melissa Scott, SEPRL Facilities Manager

SEPRL

706-546-3456, 706-614-4274 (cell)

Deputy Safety Officer (DSO), on call 4:30pm-8am

706- 546-3700

Lindsay Killmaster, IACUC Chair

706-546-3460, 706-248-0226 (cell)

Nancy Mead, Alternate Biosafety Official

706-546-3434, 706-202-1969 (cell)

Dr. David Suarez, Research Leader

EEAVDRU

706-546-3479, 706-248-9183 (cell)

Dr. John Dunn, Research Leader, EPVDRU

706-546-3642; 706-206-4212 (cell)

Todd Lonergan, IT Specialist

706-546-3975, 706-286-4867 (cell)

Guard House Emergency number 706-338-1792 RRC Guard Control Room, 6pm-6am 706-546-3536

Guard House, 6am-6pm 706-546-3112

706-546-3113

Introduction

The following biosafety/biosecurity procedures have been established at Southeast Poultry Research Laboratory (SEPRL) to prevent problems associated with the handling of biohazards.

These guidelines supersede all existing SEPRL protocols for biosafety/biosecurity; they will serve as the minimum standards of operation for all biosafety/ biosecurity at SEPRL Building 45.

These guidelines have been agreed upon by the Biosecurity Advisory Sub-Committee, a subcommittee of the USNPRC Institutional Biosafety Committee (IBC), as the appropriate procedures of operation for SEPRL at this time. This document will be reviewed periodically, subsequently; changes and updates will be made as needed.

Personnel are encouraged to discuss any questions they may have with their immediate supervisor, the Chair of the Biosecurity Advisory Sub-Committee, the Operations Director, Research Leaders and/or the Laboratory Director.

The biosafety/biosecurity procedures at SEPRL are intended to accomplish the following:

• Prevent the transfer of infectious agents to avian populations located off the

SEPRL premises. Employees are required to avoid away from work contact (except for work related reasons) with commercial poultry or other domestic or exotic avian species. This works as a critical “barrier”; an agent release from our laboratory could produce a major economic loss in the poultry industry. As a result, items, such as equipment must be adequately decontaminated before it can leave the premises. In the case of a viable material being shipped, the agent must be sufficiently contained, and the receiving laboratory must have adequate containment facilities.

• Prevent infection of personnel. The importance of preventing employee illness is accomplished with the effective use of primary barriers, good lab technique, and good personal hygiene.

• Prevent transmission of disease to SEPRL production flocks. Our specific pathogen- free (SPF) flocks are the source of chickens, turkeys and embryonated eggs for research use at SEPRL. Infection of these flocks with any of the common poultry pathogens would make eggs and progeny unsuitable for ongoing future studies.

• Prevent cross-contamination of laboratory and animal experiments. This is necessary for the production of valid and reproducible research.

General Lab - Biosafety / Biosecurity Guidelines Southeast Poultry Research Laboratory

Building 45

Facility SHEM Policy Statement

It is the policy of the U. S. Department of Agriculture, Agricultural Research Service (USDA- ARS), to safeguard its employees and property by implementing and maintaining a comprehensive, effective Safety, Health, and Environmental Management (SHEM) Program in compliance with the laws and Executive Orders. The ARS SHEM program is outlined in ARS ERS NASS NIFA Manual 160.0M that is accessible to employees on the ARS Administrative and Financial Management website. The Agency’s policy is to eliminate or minimize losses incurred by the Agency, individual employees, and the general public, as a result of actions or incidents involving or producing injury, illness property/environmental damage in the ARS workplace by:

• Implementing the Agency’s program for safety, health, and environmental management.

• Complying with the spirit and letter of safety, health, and environmental legislation and related standards, orders, rules, and regulations.

• Establishing procedures for procurement, repair, storage, waste management, disposal, and salvage operations to prevent accident-related losses.

• Providing supervisory/employee educational/training opportunities for communicating program function components.

• Properly assigning, directing, and training ARS employees.

• Developing supervisory safety, health and environmental related knowledge and skills.

• Developing facility self-protection plans for emergency situations.

• Developing and implementing program promotional activities.

• Analyzing work assignments to identify potentially hazardous conditions or adverse environmental consequences.

• Developing standard operating procedures that minimize or eliminate potentially hazardous conditions or adverse environmental effects.

• Analyzing the relationship of the employee, machine, and environment for the identification of potentially hazardous conditions or adverse environmental effects.

• Protecting employees from hazardous conditions through isolation, guarding and shielding principles or personal protective equipment.

• Developing policies and procedures that minimize or eliminate potentially hazardous conditions or adverse personal effect through chemical labeling, hygiene, proper storage, and disposal and by inventorying chemical and hazardous substances and materials.

• Developing procedures for reviewing, inspecting, auditing, and complying with the SHEM program.

The Laboratory Director (LD) has the responsibility for developing maintaining, and actively supporting a strong SHEM program within the Southeast Poultry Research Laboratory (SEPRL).

https://www.ars.usda.gov/ARSUserFiles/50000000/MWACOP/600.0(4)_May2020.pdf https://www.ars.usda.gov/ARSUserFiles/50000000/MWACOP/600.0(4)_May2020.pdf

The LD will provide administrative and managerial backing of committee recommendations and requests (e.g., training opportunities, requests for funding on installation of appropriate engineering controls, or equipment necessary for the elimination of hazardous conditions).

Security Related Policies Personnel Clearance Permanent Employees All permanent employees working at SEPRL must have appropriate clearances. Additional requirements must be met for working with Select Agents.

Non-Employees – Citizens, Visiting Scientists and Short-Term Technicians Such personnel must be escorted at all times.

Non-Citizens To work, such personnel must submit an ARS-230 form and be cleared; they must also be escorted at all times, unless other clearances are obtained.

Cameras

Neither visitors nor employees are allowed to photograph the exterior or interior of the facility buildings without permission of the Lab Director or designee.

Principles of Biosafety and Biosecurity Definitions

Biosafety - the practices, procedures, and use of equipment needed to ensure adequate safety conditions in all facilities that work with potentially infectious microorganisms and other biological hazards. “Laboratory biosafety” refers to the application of combinations of laboratory practices and procedures, laboratory facilities, safety equipment, and appropriate occupational health programs to mitigate the risks associated with handling potentially infectious microorganisms and other biological hazards (biohazards). The key principles of laboratory biosafety are hazard recognition, risk assessment, and hazard mitigation, including appropriate biocontainment.

Biosecurity - the discipline addressing the security of microbiological agents and toxins and the threats posed to human and animal health, the environment, and the economy by deliberate misuse or release.

Biohazards - infectious agents or other hazardous biologic materials that present a risk or potential risk to the health of humans, animals, or the environment. The risk can be direct through infection or indirect through damage to the environment. Biohazards include certain types of recombinant or synthetic nucleic acids; organisms and viruses infectious to humans, animals, or plants (e.g., parasites, viruses, bacteria, fungi, prions, and rickettsia);

and biologically active agents (e.g., toxins, allergens, and venoms) that can cause disease in other living organisms or cause significant impact to the environment or community.

The term biosafety/biosecurity is used in describing safe methods for managing agents in the laboratory and animal housing environment. The purpose is to reduce or eliminate exposure of personnel and the environment to potentially hazardous agents through containment of the agent being used, as well as prevent cross-contamination of facilities or spaces.

Three basic elements of containment are: 1) good laboratory practice and technique; 2) safety equipment; 3) facility design. A risk assessment of the work to be done with a specific agent will determine the appropriate combination of these elements needed.

A key to the success of the biosafety/biosecurity program is the establishment and maintenance of multiple barriers. One of the more important barriers is a change by personnel into lab supplied clothing and shoes before working with infectious materials and/or infected animals. When lab clothing is used, the individuals personal clothing does not become contaminated and serve as a fomite for carrying infectious materials away from the laboratory.

Additional precautions are added when the risk is greater, as in high containment laboratories.

and animal facilities, where a shower must be taken before leaving the building in order to prevent carrying an infective agent out of the building. Similarly, employees working in SEPRL’s clean bird Production Area must shower before they enter each flock’s room, where the concern is to avoid bringing an infective agent into our clean Production flocks.

Laboratory Practice and Technique

The most important element of containment is strict adherence to standard microbiological practices and techniques. Persons working with infectious agents or potentially infected materials must be aware of the potential hazards and be trained and proficient in the practices and techniques required to safely handle such material.

The immediate supervisor is responsible for providing or arranging for appropriate training of supervised personnel.

Safety Equipment (Primary Barriers)

Good microbiological technique and the use of appropriate safety equipment provide protection of both the personnel and the immediate laboratory environment from exposure to infectious agents.

Safety equipment includes biological safety cabinets, as well as other engineering controls designed to remove or minimize exposure of workers to hazardous biological materials and at the same time contain the agent. A biological safety cabinet is a very effective means for providing containment of splashes or aerosols generated during microbiological procedures and for protecting the worker during those procedures. Another example is the safety centrifuge cup, which is an enclosed container designed to prevent aerosols from being released during centrifugation.

Safety equipment also includes items for personal protection such as gloves, lab coats, gowns, shoe covers, boots, respirators, face shields, and safety eye protection. Personal protective equipment is often used in combination with biological safety cabinets or other devices to contain the agent or material.

In some situations, it is impractical to work in a biological safety cabinet. Personal protective equipment then becomes the primary protective barrier between personnel and the infectious material. Examples include certain animal studies, necropsy, agent production activities and activities relating to maintenance or service support of the facility.

Facility Design (Secondary Barriers)

Secondary containment - the protection of the environment outside the laboratory - is provided by a combination of facility design and operational practices.

The design of the facility is important in providing a barrier to protect persons working inside and outside the lab work-area and to protect persons and animals in the community from infectious agent releases. Laboratory management is responsible for providing facilities commensurate with the laboratory’s function and the recommended biosafety level for the agents being manipulated.

The recommended secondary barrier(s) will depend on the risk of transmission of specific agents.

Minimally, these secondary barriers include separation of the laboratory work-area from public access, availability of decontamination equipment (e.g., autoclave), and hand washing facilities.

As the risk for aerosol transmission increases, higher levels of primary containment and multiple secondary barriers may become necessary to prevent infectious agents from escaping into the environment. Design features could include specialized ventilation systems to assure directional airflow, air treatment systems to decontaminate or remove agents from exhaust air, controlled access zones within buildings or fencing to separate buildings and containment zones, airlocks at laboratory entrances, or separate buildings or modules for isolation of the agent.

No Contact with Avian Species Policy

A crucial element in SEPRL’s efforts to maintain a high level of biosecurity is dependent upon its personnel, service personnel, and visitors who have contact with SEPRL. The effectiveness of this effort depends upon an understanding of the need and commitment to SEPRL’s mission.

It is very important that employees, SEPRL’s Operation and Maintenance employees, as well as, visiting laboratory personnel, comply with the policy of having no contact with any avian species or their housing when away from SEPRL. This requirement is our first defensive effort in preventing infection of our flocks as well as guarding against the escape of an infectious agent.

As a current employee, you have read and signed the SEPRL No Off-Site Contact with Avian Species memo.

Supervisors should inform prospective employees of these avian no-contact restrictions during the interview process and be sure that they read this memo and agree to comply if employed.

New employees are required to sign a copy of this memo as they begin working at SEPRL.

Visiting laboratory personnel are also required to sign this memo and not to have any avian contact for a period of one week before and after the completion of their visit. A copy of the memo can be found on the following page.

DATE:

July 2020

SUBJECT: No Off-site Contact with Avian Species TO: SEPRL Employees, Visiting Laboratory Workers and Contractors FROM: D. E. Swayne, Laboratory Director

MEMO – No Off-Site Avian Contact Southeast Poultry Research Laboratory [SEPRL] is a unique research facility that includes a production unit for rearing specific-pathogen-free [SPF] chickens and turkeys; and containment facilities for research on exotic, endemic and emerging avian disease problems that impact avian and human health.

Disease transmission among poultry flocks is often associated with personnel or equipment movement from infected to susceptible flocks. To prevent the spread of infectious diseases at SEPRL, we rely on a series of operational [clothes changes, showers, etc.] and physical barriers. These barriers, described in detail in SEPRL’s General Laboratory Biosafety/Biosecurity Guidelines, are intended to prevent the release of infectious agents from our laboratory and animal buildings. We require these precautions for two reasons: 1] chickens, turkeys, and pet birds are susceptible to one or more of the infectious agents in our research program; 2] infections of commercial poultry & pet birds are a potential hazard to our production flocks. Consequently, we are not only concerned about what personnel might accidentally carry away, but we are also concerned about what personnel can bring to SEPRL! Such events could result in the infection of our SPF flocks or flocks outside our premises at a great economic loss. Either event would have a disastrous impact on our perceived capability & could affect our ability to continue our research program.

These restrictions may be more restrictive than at laboratories where you might have worked in the past.

But such requirements are not unique to this laboratory. The USDA Animal & Plant Health Inspection Service has a similar rule that prohibit its employees from coming into contact with susceptible species for at least seven days after being in contact with or on a premise where infected species are located.

Hopefully, everyone will understand the reason for these restrictions & will comply with not only the letter but also the spirit of the rule; thereby realizing the serious negative effects that could result from the inadvertent release of a pathogen from our laboratory.

Signature:

Date:

Printed Name:

As a final barrier is the stipulation that employees are not to come into contact with susceptible avian species or their housing when away from SEPRL, without written permission from the Lab Director [or his representative]. This prohibition includes no visit to or working on a poultry farm or other location where poultry are present; and no poultry or pet bird(s) in your residence or on the property where you reside. The Lab Director can give an exemption to limited avian contact, as long as the employees & visitors must not have any contact with susceptible animal species for a period of at least five [5] days after being in a BSL-2 lab or animal building used for endemic disease research & seven [7] days after being in a BSL-3+ lab or animal building used for exotic & emerging disease research.

Please sign, date & return this memo to the Biosafety Officer, so that we will have a record that you have received, read & understand these requirements.

Exceptions to SEPRL’s No Avian Contact

Policy

The Laboratory Director (LD) has agency and unit responsibility for biosecurity and biosafety at SEPRL. The LD can grant a temporary exception to SEPRL’s No Avian Contact for Employees Policy on a case-by-case basis; however, such exceptions will be only for research purposes related to grantee’s official work, but the time limit will not be reduced below the regulatory requirement of five (5) days for low pathogenic avian influenza or seven (7) days for SA access. SEPRL has an absolute, no time-limit, no avian contact rule for off-site for all employees.

Seeking approval for an exception to the No Avian Contact Policy:

• The decision to make an exception to the No Avian Contact Policy remains with the Laboratory Director. Exceptions to the policy are not at the discretion of the employee.

Activities that do not require approval from the LD are listed at the end of this section.

The LD may consult with the Biosecurity Advisory Committee.

• Approval for avian contact off-site is temporary and for a specified period.

• If you intend to go to a location where you might encounter contact with avian species, ask the Laboratory Director or Chair of the Biosecurity Advisory Committee about the need for approval.

• Avian contact off-site includes visits to commercial and non-commercial poultry houses and farms; hatcheries, live-poultry markets, distributors, auctions, and other activities where the employee might have direct contact with avian species.

• Employees that work in SEPRL BSL--3E areas should take extra care when entering situations where there may be unexpected bird contact (away from work).

Required conditions for approval of off-site avian contact:

• The employee must have sufficient time away from the direct handling of SEPRL infectious agents, infected materials or infected animals and not have been in a containment laboratory or animal room with infectious materials for a reasonable period before contact with avian species off-site.

This time period before contact off-site will generally be 5 days for BSL-2 agents and 7 days for BSL-3 agents.

An employee returning to SEPRL after avian contact off-site should not have direct contact with any production facilities at SEPRL, including equipment rooms, for 5-7 days.

• During the approved period for avian contact off-site, the employee should not have contact with infectious agents, infected materials, or infected animals while at SEPRL and should not enter SEPRL’s containment laboratories or animal rooms.

• Clothing, shoes, and related laboratory items that will be worn off-site should be new, laundered, clean and/or disinfected as appropriate to reduce the risk of transferring any infectious material off-site.

• Any equipment that will accompany the employee off-site must be decontaminated before leaving SEPRL.

• Returning samples and equipment must be handled as contaminated materials until appropriately decontaminated or used for research purposes in facilities at the appropriate level of biocontainment.

• Decontamination methods appropriate for materials coming back to SEPRL should be a component of the plan for off-site avian contact.

Activities that do not require approval from the Laboratory Director:

• Visits to public exhibitions, parks or recreation areas that will not involve direct avian contact, such as, zoological parks, wildlife refuges, local parks, lakes, etc.

• Post-mortem examination of avian species off-site in a teaching, diagnostic or laboratory setting provided all contact is with dead or euthanized birds and that the birds will be disposed of in a proper manner.

Facility Operating Levels SEPRL facilities include clean areas, areas used for the housing and production of the SPF flocks, and laboratory and animal areas operating at different biosafety levels. Each area requires specific work habits, personnel restrictions, and clothing stipulations.

Clean Areas

• Building 45- Offices, Hallways, Basement, Meeting Rooms.

Production Areas

• Buildings 13, 15, 16, 17, 43, and 44

Biosafety Level 2 Areas

• Building 45 - All laboratories

Animal Biosafety Level 2 Areas

• Buildings 6, 8, 39, and Russell Research Center (RRC) Building 15

Clothing Requirements and Movement of Personnel The success of our biosafety/biosecurity program depends on those who work at SEPRL. The program’s aim is to prevent any transfer of a pathogen from one experimental room to another; but it is most important that we not transfer a virus from a higher level of containment, such as Building 47 or Building 45 to a lower level or uncontaminated area, such as the Production Area or away from SEPRL.

General Information

Visitors - All

• Visitors must always be escorted.

• Exceptions to the escort policy may be given by the Laboratory Director on a case-by-case basis, but the post exposure time limit as specified by APHIS for avian contact after exposure to SEPRL birds and will not be reduced below the regulatory requirements.

Visitors – Individuals Touring Specific Areas

• Visitors who are permitted to enter an animal building or lab area are required to sign a Waiver of Liability and SEPRL No-Avian Contact document.

• Visitors who have been given permission to enter a laboratory in Building 45 Compound or Production building must change into lab-supplied clothing or Production-specific clothing.

• Visitors will be briefed on the nature of the operation to be visited and will also be briefed on the potential physical, chemical, or biological hazards for which they may be exposed, as well as any required action the visitor must take to mitigate exposure. This will be documented and covered in the BSL-2 Checklist.

• Visitors who are permitted to enter a BSL-2E building or lab area, can participate in work projects or work in progress, after proper training.

• Any personal protective equipment that is required in the area to be visited will be provided.

• Refusal to utilize required personal protective equipment will result in the denial of entry into the area.

When visitors have completed their tour, their lab-supplied clothing will be placed in the change room laundry bins or gathered in a plastic bag for incineration.

Tours

• SEPRL does not generally allow tours into the Compound or Production buildings while they are in use - entry into any building will be by permission of the LD.

• Tours will move from areas of clean status to areas of biocontainment, i.e., tours will move from the Production Area to the Compound Area.

• On special occasions, as when a tour of a specific clean area or building is being conducted, special clothing for the visitors may be required. The Laboratory Director will make this decision.

When visitors have completed their tour and returned Building 45, their lab-supplied clothing will be placed in the laundry bin or gathered in a plastic bag for incineration.

Service persons

• In general, outside service persons will be always escorted. They can be left to do their work in BSL-2 labs, if there is someone in the area, they are working in but should be “checked-on” periodically.

• The escort must inform their visitor of the location of the emergency exit and safety eyewashes/showers.

• Clear away any biohazard or chemical hazard that can be moved, so that the service person cannot come in contact with it. It is the employee’s responsibility, as an escort, to inform the service person of any hazards that cannot be removed from their work area. It is the employee’s responsibility to ensure that any equipment that is to be serviced is disinfected and free of any contaminating agents before being serviced. The SEPRL Decontamination Form can be found in the Appendix of this manual and on the Company Folder. Contact the Biosafety Officer or Biosafety Specialist if you need assistance.

SEPRL LABORATORY EQUIPMENT DECONTAMINATION FORM

• This form [see Appendix] is to be used on all equipment, regardless of whether it is to be disposed of, removed to another laboratory, or remain in the laboratory, must be disinfected and or decontaminated with an appropriate disinfectant.

• Complete and attach the SEPRL Laboratory and Equipment Decontamination Form (located at the end of this document) to each piece of decontaminated equipment.

• Routine service persons (as determined by the Laboratory Director) that have become familiar with SEPRL's BSL-2 biosecurity requirements can enter BSL-2 lab areas unescorted if their work is coordinated with BSL-2 lab personnel where the work is to be done.

• Service personnel are required to don the appropriate personal protective equipment and abide by clothing requirements for specified lab areas.

Visiting Scientists (VS), Collaborators (C) and Visiting Technicians (VT)

• Visiting scientists, collaborators and visiting technicians (VS/C/VT) must abide by the same rules as SEPRL employees. VS/C/VT are expected to spend a part of their initial training time in becoming familiar with SEPRL’s lab safety requirements and biosecurity/biosafety guidelines.

• Approval must be obtained from the LD before VS/C/VT will be permitted access and allowed to conduct research in SEPRL BSL-2 laboratories.

• VS/C/VT must be escorted by their sponsoring SEPRL Federal employee or a designee during all hours, while at SEPRL. VS/C/VT are never to be left unescorted.

General Clothing Requirements

Laundering Lab-supplied Clothing

• Compound ABSL-2E animal buildings will hold their used lab-supplied clothing/towels until experiments are completed and the building has been decontaminated. After decontamination, materials will be taken to B45 for laundering.

• Production personnel must carry their lab-supplied clothing directly to B47 on the pre-assigned day for laundering onsite. Personnel also have the option of washing their own lab-supplied clothing in the on-station washing machine.

• Lab-supplied clothing is never to be removed from SEPRL for laundering. In addition, lab-supplied clothing is never to be worn outside of SEPRL.

• Lab-supplied clothing from all BSL-2 laboratories is to be placed in the dirty clothes bin provided in B45 change rooms. All personnel, visitors included, who change clothes in any other change rooms in the compound are to place their worn lab-supplied clothing in these change room containers.

• Engineering and Operations and Maintenance (O and M) personnel lab-supplied clothing are picked up weekly and laundered onsite.

Clean Clothing

Definition: clothing that has not been worn into a Biosafety Level 2 area where it might have been exposed to infectious agents.

• Lab-supplied clean clothing is stored in the B45 Laundry Storage. Each employee is responsible for obtaining his or her needed supply of clothing from this stock.

• O and M and engineering personnel are provided with work clothing [pants and shirts or coveralls and safety shoes] to be worn as needed.

Laboratory Coats

• Wear white, front buttoning, lab coats when working in the BSL-2 laboratories.

Coveralls/Scrubs

• A choice of coveralls and scrubs are provided for employees working in the Compound and Production areas.

• Lab workers should wear scrubs [and lab coat] while working in the labs but wear scrubs with wrap-around gowns while working with infected animals. When working with uninfected animals, scrubs alone may be worn.

Safety Glasses

• Safety eye protection must be worn by all personnel when entering any lab area or animal building.

Shoes

• The purchase of laboratory safety shoes is by arrangement with your supervisor.

• The laboratory safety shoe requirement can be met so long as the shoe has a solid, non-cloth top, a heel back and a non-slip sole (non-lacing). There are several local sources, and they can also be ordered.

• Steel-toed safety shoes are required on an as-needed basis.

• Street shoes can be worn only in the B45 clean areas; street shoes can be worn in to

BSL-2 lab areas if booties are donned over street shoes.

• Building-dedicated rubber boots are to be worn in the animal rooms of the Compound and Production Areas; they are to be disinfected in a disinfecting bath when leaving each room. Rubber boots for Production areas should not be worn in other parts of the facility.

• Rubber boots can be obtained from the Facility Manager.

Undergarments/Socks

• Personnel can bring their own undergarments or have theirs purchased through their supervisor.

• A supply is also maintained as inventory in the B45 Laundry Storage.

Specific Clothing Requirements

Clean Areas

• Building 45 – Administrative and Office Areas

Building 45 – Admin and Offices Street clothing or clean laboratory clothing (scrubs) may be worn in these areas.

Building 45 - Meeting Rooms, Break Rooms, Equipment Rooms Street clothing or clean laboratory clothing may be worn in these areas. This includes when meeting with scheduled visitors, either in one’s office, attending gatherings in the meeting room or having lunch in the break room.

Production Area

Entry into any Production Area building is generally restricted to those employees assigned to the area. The Laboratory Director or designee must give permission for other employees to enter.

Engineering and O and M Personnel

• It is important that O and M personnel are conscious of their clothing and movements daily; you are more likely to be in more areas in a day than a person assigned to a particular area.

Individuals working within different areas of the compound as their job dictates must be mindful of their movements within the compound and plan their work accordingly. For biosecurity concerns, those who have been in BSL-2/3 areas should not return to the Production Area during the same day. The wait times for returning to the Production area are 5 days for BSL-2 areas (or contact with avian species) and 7 days for BSL-3 areas.

Buildings 13, 15, 16, 17, 43, 44

Employees

• A shower and hair shampoo, as well as a change into lab-supplied clothes is required before entry into any animal housing room in the Production Area.

• Compound personnel are not allowed in the Production Area, unless approved by the

LD.

• In general, before entering any of these buildings, permission must be obtained from the Laboratory Director or Biosafety/Biosecurity Officer, and the Attending Veterinarian.

• Before entering any of these buildings, a minimum of a 5-day wait is required for BSL- 2 containment area workers and a one-week wait for BSL-3E containment area workers.

• Biosecurity for Buildings 13, 15, 16, 17, 43, and 44 is critical for SEPRL’s mission.

These are limited access areas and have shower-in requirements in place in these SPF buildings. The strictest practice of biosecurity for personnel and equipment movement is to be maintained when entering these buildings.

• Building-dedicated scrubs, undergarments, rubber boots, dust mask, protective eyewear and shoe covers are provided for poultry production employees.

• If multiple buildings will be entered in a single day, the rooms must be entered in sequence, youngest to oldest flocks.

• Once a flock has been passed in the sequence by a single person by entering a subsequent flock, that person cannot reenter the preceding flocks until the following day.

Visitors

• In general, visitors are only allowed in these buildings after the building has been fumigated and in a clean status. Visitors will be provided with shoe covers or Production boots, and other appropriate clothing depending on areas visited.

• Entry into a building will be by permission of the Lab Director or designee.

• A five-day waiting period is required for individuals who have had access to avian species, or BSL-2 spaces. A seven-day waiting period is required for visitors who have accessed BSL-3 spaces.

• The Production Area must be the first stop for any facility tours.

• Visitors must be always escorted. Access is generally limited to observation windows only. Access to animal rooms requires prior approval of the LD and Attending Veterinarian. Safety glasses may be required.

• Access is allowed for observation only!

• Only ONE visitor at a time may enter a Production animal room.

• A shower and hair shampoo, as well as a change into Production-supplied clothes, is required before entry into any animal housing room in the Production Area.

• Building-dedicated scrubs, undergarments, rubber boots, dust mask, protective eyewear and shoe covers are provided for visitors entering animal rooms.

• If multiple buildings will be entered in a single day, the rooms must be entered in sequence, youngest to oldest flocks.

• Once a flock has been passed in the sequence by a single person by entering a subsequent flock, that person cannot reenter the preceding flocks until the following day.

Engineering and O and M Personnel

• A shower and hair shampoo and change into lab-supplied clothes is required before entry into any animal housing room.

• Engineering and O and M personnel must have changed into coveralls before entry into any of these buildings.

Individuals working within different areas of the compound as their job dictates must be mindful of their movements within the compound and plan their work accordingly. For biosecurity concerns, those who have been in BSL-2/3 areas should not return to the Production Area during the same day. The wait times for returning to the Production area are 5 days for BSL-2 areas (or contact with avian species) and 7 days for BSL-3 areas.

Building 45

• A change into lab-supplied clothing and approved safety shoes is required before entry into the laboratory suite. A buttoned, white lab coat must be worn along with safety glasses. The lab coat is to remain in the work-area.

Visitors

• Visitors will be provided with a clean, white lab coat, safety glasses, shoe covers and will be always escorted. If approved to enter the lab area for a work collaboration, scrubs may be required.

• Engineering and O and M personnel are required to wear lab-supplied clothing as a minimum.

• To mitigate the exposure of service personnel to any biological agents or hazardous chemicals, it is the responsibility of SEPRL personnel whose equipment is being serviced to be certain that the equipment being serviced, as well as, surrounding areas has been disinfected before the service person can touch it.

• Service persons are required to wear a clean, white lab coat and shoe covers as a minimum when going into these areas.

Biosafety Level 2 Areas

Laboratories

• A complete change into lab-supplied clothing and approved safety shoes, in the designated change rooms, is required before entry into these areas and buildings. This change of clothing includes removal of all outer clothing: pants, shirts, dresses, skirts, sweaters, and shoes [undershirts are exempt, but must be covered by a scrub shirt].

• A change into b u i l d i n g - dedicated s h o e s is required when entering the lab area.

These shoes should never leave the lab. Likewise, a change out of these shoes is required when exiting the building.

• A buttoned white laboratory coat must be worn when working in the laboratory with hazardous materials. This white lab coat is to remain in the work-area when exiting to the change room.

• Disposable plastic shoe covers are provided for workers who only occasionally go into these buildings and who do not have a pair of dedicated shoes in the building.

Visitors

• Visitors will be allowed in these buildings only by permission of the Laboratory Director or Biosafety Officer.

• Visitors must be always escorted.

• Visitors will be provided with the appropriate personal protective equipment such as lab specific clothing and shoe covers.

• A white lab coat should be put on when inside the building. The shoe coverings should also be exchanged for new shoe coverings for going into the building lab areas.

Engineering and O and M Personnel

• Engineering and O and M personnel are required to wear clean coveralls and disposable plastic shoe covers over their lab-supplied clothing when going into these buildings. These clothes will be put on and removed in the building change room.

• To mitigate the exposure of service personnel to any biological agents or hazardous chemicals, it is the responsibility of SEPRL personnel whose equipment is being serviced to be certain that the equipment being serviced, as well as, surrounding areas has been disinfected before the service person is allowed to touch it.

• Service persons will be provided with disposable or clean non-disposable coveralls and shoe covers. These clothes will be put on over their personal clothing and removed in the change room.

• A white lab coat should be put on when inside the building and removed before exiting. The shoe coverings should also be exchanged for new shoe coverings when going into the building lab areas.

Animal Housing - Buildings 6, 8, and 39 Employees

• Compound animal caretakers must change in Building 45 into lab-supplied clothing before entering the Compound Area. Other changes of outer and under clothing may be required within buildings of the Compound. A shower and clothing change out of a building may be required, dependent on the building and work or experiments in progress.

• Scrubs and wrap-around gowns are to be worn in the animal areas and in animal rooms rather than white lab coats.

• Scrubs brought into buildings where infectious disease work is being conducted are to be decontaminated in the room or bagged out of the room to be washed.

• Researchers and technicians entering Compound animal-housing buildings must have changed into lab-supplied clothing in the change rooms before entry; coveralls or a change of clothing may be required to be worn while working in these buildings, dependent on the agent used.

• A shower and hair shampoo and clothes change out of the building might be required -depending on the building and work or experiment in progress.

• Building-dedicated rubber boots are required to be worn in the animal rooms and disinfected when leaving the room.

• Compound personnel are required to enter and perform their work duties in the order of the cleanliness of the buildings; clean buildings are to be worked first.

• Workers cannot return to a clean building during the same day if they have entered a contaminated building.

• Workers are to employ the strictest practice of biosecurity for equipment movement.

• Entry into Building 6, 8, or 39 requires a one-day wait if you have been into a building operating at BSL-3E.

Note: M echanical rooms are not considered “inside the building” and do not require an additional change of clothes when being entered.

Visitors

• If a visitor is given permission to go into an in-use building, they must prepare for entry and exit just as employees are expected to do, by removal of all street clothing and a change into lab-supplied clothing in the B45 change rooms. Once in the building, a wrap-around gown may be put on over the lab clothing, dependent on the status of the room/building. A shower out of the building may be required.

• Building-dedicated rubber boots are required to be worn in the animal rooms when challenged or off-site birds are present or if the room has not been cleaned after an experiment.

• Engineering and O and M personnel entering Compound animal-housing buildings must change into lab-supplied clothing before entry; additional clothing or a change of clothing may be required to be worn while working in these buildings. A shower and clothing change out of the entered building might be required, dependent on the building and work or experiment in progress.

Note: M echanical rooms are not considered “inside the building” and do not require an additional change of clothes when entering.

• Building-dedicated rubber boots are required to be worn in the animal rooms when

• Service persons should not be exposed to any viable biological agents or hazardous chemicals. It is the responsibility of the person whose equipment is being serviced to be certain that the equipment being serviced [and its surrounding area] has been disinfected before the service person can touch it.

• Service personnel entering an in-use Compound animal-housing buildings must change into lab-supplied clothing in the Building 45 change rooms before entry;

additional clothing or a change of clothing may be required to be worn while working in these buildings. A shower and clothing change out of the building may be required, dependent on the building and work or experiments in progress.

• Building-dedicated rubber boots are required to be worn in the animal rooms when

Laboratory Equipment

Autoclaves, centrifuges, and biological safety cabinets should undergo regular preventative maintenance by qualified personnel. Preventative maintenance records must be kept for each piece of equipment. The airflow must be regularly checked on the biological safety cabinets and filters changed by qualified, NSF-certified personnel.

Regardless of use, all lab equipment used in microbiological or biomedical research should be disinfected routinely. Thorough decontamination should be provided for any equipment after a spill, splash, or potential contamination. Equipment must be decontaminated before repair, maintenance or removal from the laboratory, and a Decontamination Tag must be affixed to the equipment.

Equipment that creates a hazard must be evaluated and considered when performing a risk assessment for biohazardous materials management. Initial evaluation should be conducted by the scientific staff through a job hazard analysis (JHA). The Biosafety Office may be contacted for assistance with this process.

Autoclaves

Introduction

A properly operating autoclave is the easiest and most effective method for disinfecting contaminated waste. The time required to sterilize a load depends not only on the temperature, but on the amount of material within the chamber. Proper autoclaving technique requires the combined effect of heat and moisture. Steam penetration throughout the material is essential for complete sterilization. For this reason, containers or bags used for autoclaving waste must have a means for steam penetration. Autoclave bags that must be tightly closed prior to autoclaving should have additional water (250 mL) added to the bag to serve as a source of steam to insure complete sterilization.

Only personnel with adequate training on autoclave procedures should be permitted to operate an autoclave. It is the responsibility of the laboratorian supervisor to provide and document the training of autoclave operators. Personnel should wear appropriate personal protective equipment: fully fastened lab coat (BSL-2), heat resistant gloves, and eye protection. Autoclave operators should inspect the autoclave components for proper operations: door clamps and seals should be inspected for wear and damage and debris should be cleared from the chamber floor drain. If a problem is found, notify the Laboratory/Program supervisor immediately. Qualified autoclave operators shall understand the time, temperature, pressure relationships required for proper material decontamination. Additional training on handling materials to be decontaminated should also be provided. Supervisors should maintain a permanent record of training provided to their staff and provide a copy to the Biosafety Officer or Biosafety Specialist.

The following parameters contribute to autoclave effectiveness:

• Temperature: unless specifically instructed by media manufacturers’ directions, autoclave chamber temperature should be at least 121°C (250°F).

• Time: cycle time will vary according to the contents of the autoclave. If media is to be prepared, the manufacturers’ instructions should be followed. At SEPRL, the autoclave time for biohazardous waste is a minimum of 60 minutes, measured after the temperature of the material being sterilized reaches 121°C and 15 psi pressure. The tighter the autoclave is packed, the longer it will take to reach 121°C in the center of the load.

• Steam Contact: steam saturation of the load is essential for effective decontamination.

Air pockets or insufficient steam supply will prevent adequate saturation. To ensure adequate steam contact, leave autoclave bags partially open during autoclaving to allow steam to penetrate the bag. The addition of a small amount of water (250 mL) inside the bag before autoclaving will help ensure heat transfer to the items being decontaminated (do not add water if it will cause biohazardous materials to splash out of the bag).

• Containers: use leak-proof autoclavable containers only. Always consider substitutes for glassware when selecting containers. Plastics such as polypropylene, polypropylene copolymer or fluoropolymer products are capable of being autoclaved repeatedly. Place non-borosilicate glass bottles in a tray of water to help prevent heat shock. Place plastic bags inside a secondary container in the autoclave in case liquids leak out. Autoclavable plastic or stainless-steel containers are appropriate secondary containers. Make sure plastic bags and pans are autoclavable to avoid melting.

• Autoclave cycle verification: validation of load is required using an autoclave log and an indicator of some type. At SEPRL, a tape indicator and a chemical indicator must be run with each load.

Tape indicators are adhesive backed paper tape with heat sensitive chemical indicator markings.

These markings appear only when the tape has been exposed to normal autoclave decontamination temperatures and are not designed to prove that organisms have been killed. Tape indicators should be used on all material decontaminated by autoclaving to show that the material has been processed.

A three-to-four-inch length of tape placed on the outside of the autoclave pan, bag, or individual container is sufficient for visual confirmation by the operator.

With each load, a chemical indicator should be used to test for the presence of heat, pressure, and the presence of steam. After the run, these indicators should be attached to the daily use log.