biosafety manual 2017 - nau · 2018-08-15 · 4 environmental health & safety contact...

Biosafety Manual 2017

Revised 6/2017

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Policy Statement

It is the policy of Northern Arizona University (NAU) to provide a safe working environment. The primary

responsibility for insuring safe conduct and conditions in the laboratory resides with the principal

investigator.

The Office of Biological Safety is committed to providing up-to-date information, training, and monitoring

to the research and biomedical community concerning the safe conduct of biological, recombinant, and acute

toxin research and the handling of biological materials in accordance with all pertinent local, state and

federal regulations, guidelines, and laws. To that end, this manual is a resource, to be used in conjunction

with the CDC and NIH guidelines, the NAU Select Agent Program, Biosafety in Microbiological and

Biomedical Laboratories (BMBL), and other resource materials.

Introduction

This Biological Safety Manual is intended for use as a guidance document for researchers and clinicians

who work with biological materials. It should be used in conjunction with the Laboratory-Specific Safety

Manual, which provides more general safety information. These manuals describe policies and procedures

that are required for the safe conduct of research at NAU. The NAU Personnel Policy on Safety 5.03 also

provides guidance for safety in the workplace.

Responsibilities

In the academic research/teaching setting, the principal investigator (PI) is responsible for ensuring that

all members of the laboratory are familiar with safe research practices. In the clinical laboratory setting, the

faculty member who supervises the laboratory is responsible for safety practices.

Lab managers, supervisors, technicians and others who provide supervisory roles in laboratories and

clinical settings are responsible for overseeing the safety practices in laboratories and reporting any

problems, accidents, and spills to the appropriate faculty member.

Employees who work with biological materials are responsible for reading this manual and carrying out the

safety practices outlined here.

The Office of Biological Safety will provide guidance, information, review, monitoring, and training

regarding biological safety programs, when appropriate. This includes implementing registration activities

for certain research projects, acting as a consultant for departments regarding implementation and

enforcement of biological safety programs, evaluating work practices and personal protective equipment,

providing educational materials, tracking employee training, and medical monitoring.

The mission of the EH&S Office of Biological Safety is to minimize injury to faculty, staff, students and

visitors and to minimize damage to University property. Inherent in this mission is the charge to provide a

safe and healthy environment in which the University’s activities can be pursued.

All applicable federal and state safety laws, rules and regulations are adopted by the University. In order to

carry out its duties and responsibilities, the EH&S Office of Biological Safety will adhere to standards or

codes related to biosafety which have been adopted and promulgated by nationally recognized standards-

setting organizations. The interpretation of biosafety regulations and guidelines is the responsibility of the

EH&S Office of Biological Safety.

In order to assure an effective Biosafety Program for Northern Arizona University, it is imperative that all

individuals associated with the University comply fully with the policies and procedures set forth in the

manual.

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Emergency Phone Numbers

General

University Police Department 928-523-3000

Flagstaff Fire Department 911

Gas leak 911

Spills/Accidents

Biological or Recombinant Materials 480-248-0741

Chemicals (laboratory) 928-308-6507

Radioactive Materials 928-220-1728

Select Agents 480-248-0741

Incidents involving Air Monitoring 928-220-1728

Medical Emergency

Campus Health Services 928-523-2131

Flagstaff Medical Center Emergency 928-779-3366

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Environmental Health & Safety Contact Information

General Information Department Location: Peterson Hall, Building 22, Room 215

Mailing Address: PO Box 4137, Flagstaff, AZ 86011-4137

Email: [email protected]; [email protected]; [email protected]

Phone: 928-523-7288

Fax: 928-523-1607

To request non-emergency work, complete a service request at nau.edu/logger/orc.

Mick Kelly Shelley Jones Hazardous Waste Coordinator Director of Biological Safety

Office: 928-523-5903 Biological Safety Officer

Cell: 928-308-6507 Office: 928-523-7268

Email: [email protected] Cell: 480-248-0741

Email: [email protected]

Garett Hall Janelle Runberg Baron Environmental Compliance Technician Assistant Biological Safety Officer

Office: 928-523-1146 Office: 928-523-4782

Cell: 928-607-3059 Cell: 480-516-1072

Email: [email protected] Email: [email protected]

Jim Biddle Emily Kaufman Environmental & Industrial Hygiene Biosafety Specialist

Office: 928-523-6109 Office: 928-523-1746

Cell: 928-220-1728 Cell: 928-533-7442


Sarah Ells David Faguy Health/Safety Officer Assistant Vice President for Regulatory

Office: 928-523-3961 Office: 928-523-6117

Cell: 928-607-6857 Cell: 435-850-9048


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Table of Contents

1.0 BIOLOGICAL SAFETY ............................................................................................7 1.1 Principles of Biological Safety .............................................................................7

1.1.1 Laboratory Practice and Technique ............................................................... 7 1.1.2 Safety Equipment (Primary Barrier) .............................................................. 8 1.1.3 Facility Design (Secondary Barrier) .............................................................. 8 1.1.4 Biosafety Levels ............................................................................................ 9

1.1.5 Animal Facilities .......................................................................................... 11 1.1.6 Clinical Laboratories .................................................................................... 11 1.1.7 Importation and Interstate Shipment of Certain Biomedical Materials......... 12

1.2 Biological Safety Levels (BSLs) ........................................................................12 1.2.1 Biosafety Level 1 (BSL-1)........................................................................... 12 1.2.2 Biosafety Level 2 (BSL-2)........................................................................... 13

1.2.3 Biosafety Level 3 (BSL-3)........................................................................... 17 1.2.4 Animal Biosafety Level 1 (ABSL-1) ........................................................... 21 1.2.5 Animal Biosafety Level 2 (ABSL-2) ........................................................... 23 1.2.6 Animal Biosafety Level 3 (ABSL-3) ........................................................... 26

1.3 Agents List ........................................................................................................33

1.3.1 Risk Group 1 (RG1) Agents ........................................................................ 33 1.3.2 Risk Group 2 (RG2) Agents ........................................................................ 33 1.3.3 Risk Group 3 (RG3) Agents ........................................................................ 38 1.3.4 Risk Group 4 (RG4) Agents ........................................................................ 39

2.0 INFORMATION FOR RESEARCHERS ...............................................................40 2.1 Project Registration ...........................................................................................40

2.1.1 Biological Agent (BA) Registration ............................................................. 41 2.1.2 Recombinant DNA (RD) Registration ......................................................... 41 2.1.3 Acute Toxins (AT) Registration .................................................................. 42 2.1.4 Project Amendments ................................................................................... 42 2.1.5 Select Agents ............................................................................................... 44

2.2 Minors in Research Laboratories or Animal Facilities .......................................46

2.3 Principle Investigator’s Responsibilities During the Conduct of Research ........47 2.3.1 Reportable Incidents and Violations ............................................................ 47 2.3.2 Institutional Reporting Responsibilities ....................................................... 48

2.4 Biological Waste Disposal Policy ......................................................................48 2.4.1 Categories .................................................................................................... 49 2.4.2 Packaging .................................................................................................... 50

2.4.3 Labeling ....................................................................................................... 50 2.4.4 Transport ..................................................................................................... 50 2.4.5 Training ....................................................................................................... 51

2.5 Biological Waste Disposal Containers ...............................................................51 2.5.1 Bulk Biohazard Receptacles ........................................................................ 51 2.5.2 Sharps Boxes ............................................................................................... 52

2.5.3 Biohazard Bags ........................................................................................... 52

2.6 Autoclave Use and Testing ................................................................................52

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2.7 Disinfectants ......................................................................................................52 2.7.1 Liquids......................................................................................................... 53 2.7.2 Gasses ......................................................................................................... 55

2.7.3 Irradiation .................................................................................................... 56 2.7.4 Disinfectants Bibliography........................................................................... 56

2.8 Shipment of Biological Materials.......................................................................61 2.9 Laboratory Spills ...............................................................................................61

2.9.1 Spill in the Open Laboratory ........................................................................ 61 2.9.2 Spill in a Biological Safety Cabinet.............................................................. 62

2.10 Biological Safety Cabinets .................................................................................63 2.10.1 Biological Safety Cabinets (BSCs) .............................................................. 63 2.10.2 Horizontal Laminar Flow “Clean Bench” .................................................... 65 2.10.3 Vertical Laminar Flow “Clean Bench” ........................................................ 65 2.10.4 Operations within a Class II BSC ................................................................ 65

2.10.5 Decontamination .......................................................................................... 66

2.10.6 Engineering Requirements ........................................................................... 67

3.0 PROGRAMS ..............................................................................................................68 3.1 Occupational Health and Safety (Occupational Health Program) ......................68 3.2 Bloodborne Pathogen Program ..........................................................................69

4.0 MEDICAL SURVEILLANCE .................................................................................69

4.1 Immunoprophylaxis ...........................................................................................69 4.1.1 Prophylactic Recommendations for Working with Infectious Agents .......... 70 4.1.2 Health Surveillance for Personnel Working with Infectious Agents ............ 72 4.1.3 Blood Serum Sampling ................................................................................ 72 4.1.4 Health Assessments ..................................................................................... 72

4.1.5 Exposure to Mycobacterium Tuberculosis .................................................. 72 List of Tables

Table 1: Summary of Recommended Biosafety Levels for Infectious Agents ........................... 31

Table 2: Summary of Recommended Biosafety Levels for Activities in Which Experimentally or

Naturally Infected Vertebrate Animals Are Used .............................................................. 32

Table 3: Toxin Table ................................................................................................................. 43

Table 4: Dilutions of Household Bleach .................................................................................... 53

Table 5: Summary and Comparison of Liquid Disinfectants (Page 1) ....................................... 57

Table 6: Summary of Practical Disinfectants ............................................................................. 59

Table 7: Reprocessing Methods for Equipment Used in the Health Care Setting ...................... 60

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1.0 BIOLOGICAL SAFETY

1.1 Principles of Biological Safety

The following is from Biosafety in Microbiological and Biomedical Laboratories, 2009, HHS publication No. (CDC) 21-1112,

Centers for Disease Control & Prevention/National Institutes of Health

The term “containment” is used in describing safe methods for managing infectious agents in the

laboratory environment where they are being handled or maintained. The purpose of containment is

to reduce or eliminate exposure of laboratory workers, other persons, and the outside environment

to potentially hazardous agents.

Both good microbiological technique and the use of appropriate safety equipment provide primary

containment, the protection of personnel and the immediate laboratory environment from exposure

to infectious agents. The use of vaccines may provide an increased level of personal protection.

Secondary containment, the protection of the environment external to the laboratory from exposure

to infectious materials, is provided by a combination of facility design and operational practices.

Therefore, the three elements of containment include laboratory practice and technique, safety

equipment, and facility design. The risk assessment of the work to be done with a specific agent

will determine the appropriate combination of these elements.

1.1.1 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 infectious materials

must be aware of potential hazards, and must be trained and proficient in the practices and

techniques required for handling such material safely. The director or person in charge of the

laboratory is responsible for providing or arranging for appropriate training of personnel.

Each laboratory should develop or adopt a biosafety or operations manual that identifies the

hazards that will or may be encountered, and which specifies practices and procedures designed to

minimize or eliminate risks. Personnel should be advised of special hazards and should be required

to read and to follow the required practices and procedures. A scientist trained and knowledgeable

in appropriate laboratory techniques, safety procedures, and hazards associated with handling

infectious agents must direct laboratory activities.

When standard laboratory practices are not sufficient to control the hazard associated with a

particular agent or laboratory procedure, additional measures may be needed. The laboratory

director is responsible for selecting additional safety practices, which must be in keeping with the

hazard associated with the agent or procedure.

Laboratory personnel, safety practices, and techniques must be supplemented by appropriate

facility design and engineering features, safety equipment, and management practices.

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1.1.2 Safety Equipment (Primary Barrier)

Safety equipment includes biological safety cabinets (BSCs), enclosed containers, and other

engineering controls designed to remove or minimize exposures to hazardous biological materials.

The biological safety cabinet (BSC) is the principal device used to provide containment of

infectious splashes or aerosols generated by many microbiological procedures. Three types of

BSCs (Class I, II, III) used in microbiological laboratories are described in Section 2. Open-fronted

Class I and Class II BSCs are primary barriers which offer significant levels of protection to

laboratory personnel and to the environment when used with good microbiological techniques. The

Class II BSC also provides protection from external contamination of the materials (e.g., cell

cultures, microbiological stocks) being manipulated inside the cabinet. The gas-tight Class III BSC

provides the highest attainable level of protection to personnel and the environment.

An example of another primary barrier is the safety centrifuge cup, an enclosed container designed

to prevent aerosols from being released during centrifugation. To minimize this hazard,

containment controls such as BSCs or centrifuge cups must be used for handling infectious agents

that can be transmitted through the aerosol route of exposure.

Safety equipment also may include items for personal protection such as gloves, coats, gowns,

shoe covers, boots, respirators, face shields, safety glasses, or goggles. Personal protective

equipment is often used in combination with biological safety cabinets and other devices that

contain the agents, animals, or materials being worked with. In some situations, which it is

impractical to work in biological safety cabinets, personal protective equipment may form the

primary barrier between personnel and the infectious materials. Examples include certain animal

studies, animal necropsy, agent production activities, and activities relating to maintenance,

service, or support of the laboratory facility.

1.1.3 Facility Design (Secondary Barrier)

The design of the facility is important in providing a barrier to protect persons working inside and

outside of the laboratory within the facility, and to protect persons or animals in the community

from infectious agents that may be accidentally released from the laboratory. Facilities must be

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.

For example, the exposure risks for most laboratory work in Biosafety Level 1 and 2 facilities will

be direct contact with the agents, or inadvertent contact exposures through contaminated work

environments. Secondary barriers in these laboratories may include separation of the laboratory

work area from public access, availability of a decontamination facility (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. Such design features could include specialized ventilation systems to assure

directional air flow, air treatment systems to decontaminate or remove agents from exhaust air,

controlled access zones, airlocks as laboratory entrances, or separate buildings or modules for

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isolation of the laboratory. Design engineers for laboratories may refer to specific ventilation

recommendations as found in the Applications Handbook for Heating, Ventilation, and Air-

conditioning (HVAC) published by the American Society of Heating, Refrigerating, and Air-

conditioning Engineers (ASHRAE).

1.1.4 Biosafety Levels

Four biosafety levels (BSLs) are described, which are differentiated through the unique

combinations of laboratory practices and techniques, safety equipment, and laboratory facilities that

are required for each level. Each combination is specifically appropriate for the operations

performed, the documented or suspected routes of transmission of the infectious agents, and for the

laboratory function or activity.

The recommended BSLs for various organisms represent those conditions under which the agent

can ordinarily be safely handled. The Principle Investigator (PI) is specifically and primarily

responsible for assessing risks and for appropriately applying the recommended biosafety level

precautions. Generally, work with known agents should be conducted at the biosafety level

recommended. When specific information is available to suggest that virulence, pathogenicity,

antibiotic resistance patterns, vaccine and treatment availability, or other factors are significantly

altered, more (or less) stringent practices may be specified.

Biosafety Level 1 practices, safety equipment, and facilities are appropriate for undergraduate and

secondary educational training and teaching laboratories, and for other facilities in which work is

done with defined and characterized strains of viable microorganisms not known to cause disease

in healthy adult humans. Bacillus subtilis, Naegleria gruberi, and infectious canine hepatitis virus

are representative of microorganisms meeting these criteria. Many agents not ordinarily associated

with disease processes in humans are, however, opportunistic pathogens and may cause infection

in the young, the aged, and immunodeficient or immunosuppressed individuals. Vaccine strains

which have undergone multiple in vivo passages should not be considered avirulent simply because

they are vaccine strains.

Biosafety Level 1 represents a basic level of containment that relies on standard microbiological

practices with no special primary or secondary barriers recommended, other than a sink for hand

washing, a universal biohazard symbol posted at laboratory entrances, and an integrated pest

management program.

Biosafety Level 2 practices, equipment, and facilities are applicable to clinical, diagnostic,

teaching, and other facilities in which work is done with the broad spectrum of indigenous

moderate-risk agents present in the community and associated with human disease of varying

severity. With good microbiological techniques, these agents can be used safely in activities

conducted on the open bench, provided the potential for producing splashes or aerosols is low.

Salmonellae, clinical samples of Hepatitis B, and Toxoplasma spp. are representative of

microorganisms assigned to this containment level. Biosafety Level 2 is appropriate when work is

done with any human-derived blood, body fluids, or tissues where the presence of an infectious

agent may be unknown. Laboratory personnel working with human-derived materials should refer

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to the Bloodborne Pathogen Standard for specific, required precautions and contact NAU’s Office

of Biological Safety to receive Bloodborne Pathogen training.

Primary hazards to personnel working with these agents include accidental

percutaneous or mucous membrane exposures, or ingestion of infectious

materials. Extreme precaution with contaminated needles or sharp

instruments must be emphasized. Even though organisms routinely

manipulated at BSL-2 are not known to be transmissible by the aerosol

route, procedures with aerosol or high splash potential that may increase the

risk of such personnel exposure must be conducted within a physical containment device such as a

BSC or safety centrifuge cups. Other primary barriers should be used, as appropriate, such as

splash shields, face protection, gowns, and gloves.

Secondary barriers such as hand washing and waste decontamination facilities must be available to

reduce potential environmental contamination. Laboratory doors should be self-closing and have

locks in accordance with institutional policies.

Biosafety Level 3 practices, safety equipment, and facilities are applicable to clinical, diagnostic,

teaching, research, or production facilities in which work is done with indigenous or exotic agents

with a potential for respiratory transmission, and which may cause serious and potentially lethal

infection. Hepatitis B. (propagation of high concentrations only), Mycobacterium tuberculosis,

Coccidioides spp., and Coxiella burnetii are representative of microorganisms assigned to this

level. Primary hazards to personnel working with these agents include autoinoculation, ingestion,

and exposure to infectious aerosols.

At Biosafety Level 3, greater emphasis is placed on primary and secondary barriers to protect

personnel in contiguous areas, the community, and the environment from exposure to potentially

infectious aerosols. For example, all laboratory manipulations should be performed in a BSC or

other enclosed equipment. Secondary barriers for this level include controlled access to the

laboratory and a specialized ventilation system that minimizes the release of infectious aerosols

from the laboratory.

Biosafety Level 4 practices, safety equipment, and facilities are applicable for work with

dangerous and exotic agents that pose a high individual risk of life-threatening disease, which may

be transmitted via the aerosol route, and for which there is no available vaccine or therapy.

Additionally, agents with a close or identical antigenic relationship to Biosafety Level 4 agents

should also be handled at this level. Viruses such as Marburg or Congo-Crimean hemorrhagic

fever are manipulated at Biosafety Level 4.

The primary hazards to personnel working with Biosafety Level 4 agents are respiratory exposure

to infectious aerosols, mucous membrane exposure to infectious droplets, and autoinoculation. All

manipulations of potentially infectious diagnostic materials, isolates, and naturally or

experimentally infected animals pose a high risk of exposure and infection to laboratory personnel,

the community, and the environment.

The laboratory worker's complete isolation of aerosolized infectious materials is accomplished

primarily by working in a Class III BSC or a full-body, air-supplied positive-pressure personnel

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suit. The Biosafety Level 4 facility itself is generally a separate building or completely isolated

zone with complex, specialized ventilation and waste management systems to prevent release of

viable agents to the environment.

1.1.5 Animal Facilities

Four animal biosafety levels (ABSLs) are described for activities involving infectious disease work

with experimental vertebrates. These four combinations of practices, safety equipment, and

facilities are designated Animal Biosafety Levels 1, 2, 3, and 4; and in general, the biosafety level

recommended for working with an infectious agent in vivo and in vitro is comparable.

1.1.6 Clinical Laboratories

Clinical laboratories, especially those in health care facilities, receive clinical specimens with

requests for a variety of diagnostic and clinical support services. Typically, the infectious nature of

clinical material is unknown, and specimens are often submitted with a broad request for

microbiological examination for multiple agents (e.g., sputa submitted for “routine,” acid-fast, and

fungal cultures). It is the responsibility of the laboratory director to establish standard procedures in

the laboratory that realistically address the issue of the infectious hazard of clinical specimens.

Except in extraordinary circumstances (e.g., suspected hemorrhagic fever), the initial processing of

clinical specimens and identification of isolates can be done safely at Biosafety Level 2, the

recommended level for work with bloodborne pathogens such as hepatitis B virus and HIV. The

containment elements described in Biosafety Level 2 are consistent with the Occupational

Exposure to Bloodborne Pathogens Standard from the Occupational Safety and Health

Administration (OSHA), which requires the use of specific precautions with all clinical specimens

of blood or other potentially infectious material (Universal Precautions). Additionally, other

recommendations specific for clinical laboratories may be obtained from the Clinical and

Laboratory Standards Institute (CLSI).

Biosafety Level 2 recommendations and OSHA requirements focus on the prevention of

percutaneous and mucous membrane exposures to clinical material. Primary barriers such as BSCs

(Class I or II) should be used when performing procedures that might cause splashing, spraying, or

splattering of droplets. BSCs should also be used for the initial processing of clinical specimens

when the nature of the test requested or other information is suggestive that an agent readily

transmissible by infectious aerosols is likely to be present (e.g., M. tuberculosis), or when the use

of a BSC (Class II) is indicated to protect the integrity of the specimen.

The segregation of clinical laboratory functions and limiting or restricting access to such areas is

the responsibility of the laboratory director. It is also the director's responsibility to establish

standard, written procedures that address the potential hazards and the required precautions to be

implemented.

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1.1.7 Importation and Interstate Shipment of Certain Biomedical Materials

The importation of etiologic agents and vectors of human diseases is subject to the requirements of

the Public Health Service Foreign Quarantine regulations. Companion regulations of the Public

Health Service and the Department of Transportation specify packaging, labeling, and shipping

requirements for etiologic agents and diagnostic specimens shipped in interstate commerce.

The U.S. Department of Agriculture (USDA) regulates the importation and interstate shipment of

animal pathogens and prohibits the importation, possession, or use of certain exotic animal disease

agents that pose a serious disease threat to domestic livestock and poultry. USDA/APHIS permits

are required for both importation and within USA transport of many hazardous biological agents.

1.2 Biological Safety Levels (BSLs)

The following is from Biosafety in Microbiological and Biomedical Laboratories, 2009, HHS publication No. (CDC) 21-1112,

Centers for Disease Control & Prevention/National Institutes of Health

1.2.1 Biosafety Level 1 (BSL-1)

Biosafety Level 1 is suitable for work involving well-characterized agents not known to cause

disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the

environment. The laboratory is not necessarily separated from the general traffic patterns in the

building. Work is generally conducted on open bench tops using standard microbiological

practices. Special containment equipment or facility design is not required nor generally used.

Laboratory personnel have specific training in the procedures conducted in the laboratory and are

supervised by a scientist with general training in microbiology or a related science.

The following standard and special practices, safety equipment, and facilities apply to the handling

of agents assigned to Biosafety Level 1:

A. Standard Microbiological Practices

1. Access to the laboratory is limited or restricted at the discretion of the laboratory

director when experiments or work with cultures and specimens are in progress.

2. Persons wash their hands after they handle viable materials and animals, after

removing gloves, and before leaving the laboratory.

3. Eating, drinking, use of tobacco products, handling

contact lenses, and applying cosmetics are not

permitted in the work areas where there is reasonable

likelihood of exposure to potentially infectious

materials. Persons who wear contact lenses in

laboratories should also wear goggles or a face shield. Food is stored outside the

work area in cabinets or refrigerators designated and used for that purpose only.

4. Mouth pipetting is prohibited; mechanical pipetting devices are used.

5. All procedures are performed carefully to minimize the creation of splashes or

aerosols.

6. Work surfaces are decontaminated at least once a day and after any spill of viable

material.

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7. All cultures, stocks, and regulated wastes are decontaminated before disposal by an

approved decontamination method, such as autoclaving. Materials to be

decontaminated outside of the immediate laboratory are to be placed in a durable,

leak-proof container and closed for transport from the laboratory. Materials to be

decontaminated off-site from the laboratory are packaged in accordance with

applicable local, state, and federal regulations, before removal from the facility.

8. An insect and rodent control program is in effect.

B. Special Practices: None

C. Safety Equipment (Primary Barriers)

1. Special containment devices or equipment such as a biological safety cabinet are

generally not required for manipulations of agents assigned to Biosafety Level 1.

2. It is recommended that laboratory coats, gowns, or uniforms are worn to prevent

contamination or soiling of street clothes.

3. Gloves should be worn if the skin on the hands is broken or if a rash exists.

4. Protective eyewear should be worn for anticipated splashes of microorganisms or

other hazardous materials to the face.

D. Laboratory Facilities (Secondary Barriers)

1. Each laboratory contains a sink for hand washing.

2. The laboratory is designed so that it can be easily cleaned. Walls and floors must be

constructed of water impervious materials that will stand up to hard disinfectants.

Carpeting is not allowed in laboratories.

3. Bench tops are impervious to water and resistant to acids, alkalis, organic solvents,

and moderate heat.

4. Laboratory furniture is sturdy and should not be cloth upholstered. Spaces between

benches, cabinets, and equipment are accessible for cleaning.

5. If the laboratory has windows that open, they are fitted with fly screens.


Biosafety Level 2 is similar to Level 1 and is suitable for work involving agents of moderate

potential hazard to personnel and the environment. It differs in four ways.

1. Laboratory personnel must have specific training in handling pathogenic agents and

should be directed by competent scientists.

2. Access to the laboratory must be limited when work is being conducted.

3. Extreme precautions must be taken with contaminated sharp items.

4. Certain procedures in which infectious aerosols or splashes may be created must be

conducted in biological safety cabinets or other physical containment equipment.

The following standard and special practices, safety equipment, and facilities apply to the use of

agents assigned to Biosafety Level 2:


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director when experiments are in progress.

2. Persons wash their hands after they handle viable materials and animals, after

removing gloves, and before leaving the laboratory.

3. Eating, drinking, the use of tobacco products, handling contact lenses, and applying

cosmetics are not permitted in the work areas. Persons who wear contact lenses in

laboratories should also wear goggles or a face shield. Food is stored outside the

work area in cabinets or refrigerators designated for that purpose only.


5. All procedures are performed carefully to minimize the creation of splashes or

aerosols.

6. Work surfaces are decontaminated at least once a day and after any spill of viable

material.




leak-proof container and closed for transport from the laboratory. Materials to be

decontaminated off-site from the laboratory are packaged in accordance with

applicable local, state, and federal regulations, before removal from the facility.


B. Special Practices

1. Access to the laboratory is limited or restricted by the laboratory director when

work with infectious agents is in progress. In general, persons who are at increased

risk of acquiring infection or for whom infection may be unusually hazardous are

not allowed in the laboratory or animal rooms. For example, persons who are

immunocompromised or immunosuppressed may be at risk of acquiring infections.

The laboratory director has the final responsibility for assessing each circumstance

and determining who may enter or work in the laboratory.

2. The laboratory director establishes policies and procedures whereby only persons

who have been advised of the potential hazard and meet specific entry requirements

(e.g., immunization) enter the laboratory or animal rooms.

3. When the infectious agent(s) in use in the laboratory require special provisions or

entry (e.g., immunization), a hazard warning sign incorporating the universal

biohazard symbol is posted on the access door to the laboratory work area. The

hazard warning sign identifies the infectious agent, lists the name and telephone

number of the laboratory director or other responsible person(s), and indicates the

special requirement(s) for entering the laboratory.

4. Laboratory personnel receive appropriate immunizations or tests for the agents

handled or potentially present in the laboratory (e.g., hepatitis B vaccine or TB skin

testing).

5. When appropriate, considering the agent(s) handled, baseline serum samples for

laboratory and other at-risk personnel are collected and stored. Additional serum

specimens may be collected periodically, depending on the agents handled or the

function of the facility.

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6. A Biosafety Manual is prepared or adopted. Personnel are advised of special

hazards and are required to read and to follow instructions on practices and

procedures.

7. Laboratory personnel receive appropriate training on the potential hazards

associated with the work involved, the necessary precautions to prevent exposures,

and the exposure evaluation procedures. Personnel receive annual updates, or

additional training as necessary for procedural or policy changes.

8. A high degree of precaution must always be taken with any contaminated sharp

items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels.

Needles and syringes or other sharp instruments should be restricted in the

laboratory for use only when there is no alternative, such as parenteral injection,

phlebotomy, or aspiration of fluids from laboratory animals and diaphragm bottles.

Plastic ware should be substituted for glassware whenever possible.

a. Only needle-locking syringes or disposable syringe-needle units (i.e., needle

is integral to the syringe) are used for injection or aspiration of infectious

materials. Used disposable needles must not be bent, sheared, broken,

recapped, removed from disposable syringes, or otherwise manipulated by

hand before disposal; rather, they must be carefully placed in conveniently

located puncture-resistant containers used for sharps disposal. Non-

disposable sharps must be placed in a hard-walled container for transport to

a processing area for decontamination, preferable by autoclaving.

b. Syringes that re-sheathe the needle, needle-less systems, and other safe

devices should be used when appropriate.

c. Broken glassware must not be handled directly by hand, but must be

removed by mechanical means such as a brush and dustpan, tongs, or

forceps. Containers of contaminated needles, sharp equipment, and broken

glass are decontaminated before disposal, according to any local, state, or

federal regulations.

9. Cultures, tissues, or specimens of body fluids are placed in a container that prevents

leakage during collection, handling, processing, storage, transport, or shipping.

10. Laboratory equipment and work surfaces should be decontaminated with an

appropriate disinfectant on a routine basis, after work with infectious materials is

finished, and especially after overt spills, splashes, or other contamination by

infectious materials. Contaminated equipment must be decontaminated according to

any local, state, or federal regulations before it is sent for repair or maintenance.

Equipment must also be decontaminated before removal from the facility when it

must be packaged for transport. Packaging and shipment shall be in accordance

with applicable local, state, or federal regulations.

11. Spills and accidents that result in overt exposures to infectious materials are

immediately reported to the laboratory director. Medical evaluation, surveillance,

and treatment are provided as appropriate and written records are maintained.

12. Animals not involved in work being performed aren’t permitted in the lab.


1. Properly maintained biological safety cabinets, preferably Class II, or other

appropriate personal protective equipment or physical containment devices are used

under the following conditions.

16

a. Use biological safety cabinets whenever procedures with a potential for

creating infectious aerosols or splashes are conducted. These may include

centrifuging, grinding, blending, vigorous shaking or mixing, sonic

disruption, opening containers of infectious materials whose internal

pressures may be different from ambient pressures, inoculating animals

intranasally, and harvesting infected tissues from animals or eggs.

b. Use biological safety cabinets whenever high concentrations or large

volumes of infectious agents are used. Such materials may be centrifuged in

the open laboratory if sealed rotor heads or centrifuge safety cups are used,

and if these rotors or safety cups are opened only in a biological safety

cabinet.

2. Face protection (goggles, mask, face shield or other splatter guards) is required to

prevent splashes or sprays of infectious or other hazardous materials to the face,

when the microorganisms must be manipulated outside the BSC.

3. Protective laboratory coats, gowns, smocks, or uniforms designated for lab use are

worn while in the laboratory. This protective clothing is removed and left in the

laboratory before leaving for non-laboratory areas (e.g., cafeteria, library, and

administrative offices). All protective clothing is either disposed of in the laboratory

or laundered by the institution; personnel should never take it home.

4. Gloves are required when handling infected animals and when hands may contact

infectious materials, contaminated surfaces or equipment. Wearing two pairs of

gloves may be appropriate; if a spill or splatter occurs, the hand will be protected

after the contaminated glove is removed. Gloves are disposed of when

contaminated, removed when work with infectious materials is completed, and are

never worn outside the laboratory. Disposable gloves are not washed or reused.


1. Each laboratory contains a sink for hand washing.

2. The laboratory is designed so that it can be easily cleaned. Walls and floors should

be smooth, hard and constructed of water impervious material that will stand up to

harsh disinfectants. Carpeting is not allowed in laboratory facilities.

3. Bench tops should be smooth, hard and impervious to water and resistant to acids,

alkalis, organic solvents, and moderate heat.

4. Laboratory furniture is sturdy, and spaces between benches, cabinets, and

equipment are accessible for cleaning. No cloth or fabric seating is permitted.

5. If the laboratory has windows that open, they are fitted with fly screens.

6. A method for decontamination of infectious or regulated laboratory wastes is

available (e.g., autoclave, chemical disinfection, incinerator, or other approved

decontamination system).

7. An eyewash/safety shower facility is readily available.

8. All laboratories require single pass air that is not then recirculated to any other area

of the facility. Laboratories should be negative pressure to surrounding areas to

prevent accidental spread of potentially infectious or recombinant agents.

9. Install biological safety cabinets in such a manner that fluctuations of the room

supply and exhaust air do not cause the biological safety cabinets to operate outside

their parameters for containment. Locate biological safety cabinets away from

doors, from windows that can be opened, from heavily traveled laboratory areas,

17

and from other potentially disruptive equipment so as to maintain the biological

safety cabinets’ air flow parameters for containment.


Biosafety Level 3 is applicable to clinical, diagnostic, teaching, research, or production facilities in

which work is done with indigenous or exotic agents which may cause serious or potentially lethal

disease as a result of exposure by the inhalation route. Laboratory personnel have specific training

in handling pathogenic and potentially lethal agents, and are supervised by competent scientists

who are experienced in working with these agents.

All procedures involving the manipulation of infectious materials are conducted within biological

safety cabinets or other physical containment devices, or by personnel wearing appropriate

personal protective clothing and equipment. The laboratory has special engineering and design

features.

It is recognized, however, that many existing facilities may not have all the facility safeguards

recommended for Biosafety Level 3 (e.g., access zone, sealed penetrations, directional airflow,

etc.). In these circumstances, acceptable safety may be achieved for routine or repetitive operations

(e.g., diagnostic procedures involving the propagation of an agent for identification, typing, and

susceptibility testing) in Biosafety Level 2 facilities. However, the recommended Standard

Microbiological Practices, Special Practices, and Safety Equipment for Biosafety Level 3 must be

rigorously followed. The decision to implement this modification of Biosafety Level 3

recommendations should be made only by the lab director.

The following standard and special safety practices, equipment, and facilities apply to the use of

agents assigned to Biosafety Level 3:



director when experiments are in progress.

2. Persons wash their hands after handling infectious materials and animals, after

removing gloves, and when they leave the laboratory.

3. Eating, drinking, the use of tobacco products, handling contact lenses and applying

cosmetics or lip balm are not permitted in the laboratory. Persons who wear contact

lenses in laboratories should also wear goggles or a face shield. Food is stored

outside the work area in cabinets or refrigerators designated for this purpose only.


5. Policies for the safe handling of sharps are instituted.

6. All procedures are performed carefully to minimize the creation of aerosols.

7. Work surfaces are decontaminated at least once a day and immediately after any

spill of viable material.




leak-proof container and closed for transport from the laboratory. Infectious wastes

18

from BSL-3 laboratories must be decontaminated before removal for off-site

disposal.




1. Laboratory doors are kept closed at all times.

2. The laboratory director controls access to the laboratory and restricts access to

persons whose presence is required for program or support purposes. For example,

persons who are immunocompromised or immunosuppressed, or for whom

infection may be unusually hazardous, are not allowed in the laboratory or animal

rooms. The director has the final responsibility for assessing each circumstance and

determining who may enter or work in the laboratory. No minors are allowed in

level 3 laboratories.

3. The laboratory director establishes policies and procedures whereby only persons

who have been advised of the potential biohazard, who meet any specific entry

requirements (e.g., immunizations), and who comply with all entry and exit

procedures, enter the laboratory or animal rooms.

4. When infectious materials or infected animals are present in the laboratory or

containment module, a hazard warning sign incorporating the universal biohazard

symbol, is posted on all laboratory and animal room access doors. The hazard

warning sign identifies the agent, lists the name and telephone number of the

laboratory director or other responsible person(s), and indicates any special

requirements for entering the laboratory, such as the need for immunizations,

respirators, or other personal protective measures.

5. Laboratory personnel receive the appropriate immunizations or tests for the agents

handled or potentially present in the laboratory (e.g., hepatitis B vaccine or TB skin

testing and periodic testing).

6. Baseline serum samples are collected and stored for all laboratory and other at-risk

personnel. Additional serum specimens may be collected periodically, depending on

the agents handled or the function of the laboratory.

7. A Biosafety Manual specific to the laboratory is prepared or adopted. Personnel are

advised of special hazards and are required to read and to follow instructions on

practices and procedures. An SOP manual is prepared to cover all procedures and

activities.

8. Laboratory personnel receive appropriate training on the potential hazards

associated with the work involved, the necessary precautions to prevent exposures,

and the exposure evaluation procedures. Personnel receive annual updates, or

additional training as necessary for procedural changes.

9. The laboratory director is responsible for insuring that all personnel demonstrate

proficiency in standard microbiological practices and techniques, and in the practice

and operations specific to the laboratory facility, before working with organisms at

Biosafety Level 3. This might include prior experience in handling human

pathogens or cell cultures, or a specific training program provided by the laboratory

director or other competent scientist proficient in safe microbiological practices and

techniques.

19

10. A high degree of precaution must always be taken with all contaminated sharp

items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels.

Needles and syringes or other sharp instruments should be restricted in the

laboratory for use only when there is no alternative, such as parenteral injection,

phlebotomy, or aspiration of fluids from laboratory animal and diaphragm bottles.

Plastic ware should be substituted for glassware whenever possible.

a. Only needle-locking syringes or disposable syringe needle units (i.e., needle

is integral to the syringe) are used for injection or aspiration of infectious

materials. Used disposable needles must not be bent, sheared, broken,

recapped, removed from disposable syringes, or otherwise manipulated by

hand before disposal; rather, they must be carefully placed in conveniently

located puncture-resistant containers used for sharps disposal. Non-

disposable sharps must be placed in a hard-walled container for transport to

a processing area for decontamination, preferably by autoclaving.

b. Syringes that re-sheathe the needle, needle-less systems, and other safe


c. Broken glassware must not be handled directly by hand, but must be

removed by mechanical means such as a brush and dustpan, tongs, or

forceps. Containers of contaminated needles, sharp equipment, and broken

glass should be decontaminated before disposal, in accordance with any

local, state, or federal regulations.

11. All manipulations involving infectious materials are conducted in biological safety

cabinets or other physical containment devices within the containment module. No

work in open vessels is conducted on the open bench.

12. Laboratory equipment and work surfaces should be decontaminated with an

appropriate disinfectant on a routine basis, after work with infectious materials is

finished. It must be cleaned and decontaminated after overt spills, splashes, or other

contamination with infectious materials. Contaminated equipment should also be

decontaminated before it is sent for repair or maintenance. An Equipment Release

Form, available on the EH&S website, should be used to assure that equipment has

been properly decontaminated prior to servicing. In addition, it must be packaged

for transport in accordance with applicable local, state, or federal regulations,

before removal from the facility. Plastic-backed paper toweling can be used on non-

perforated work surfaces within biological safety cabinets to facilitate clean up.

13. Cultures, tissues, or specimens of body fluids are placed in a container that prevents

leakage during collection, handling, processing, storage, transport, or shipping.

14. All potentially contaminated materials (e.g., gloves, lab coats, etc.) from

laboratories or animal rooms are decontaminated before disposal or reuse.

15. Spills of infectious materials are decontaminated, contained and cleaned up by

appropriate professional staff, or others properly trained and equipped to work with

concentrated infectious material.

16. Spills and accidents that result in overt or potential exposures to infectious materials

are immediately reported to the laboratory director. Appropriate medical evaluation,

surveillance, and treatment are provided and written records are maintained.

17. Animals and plants not related to the work being conducted are not permitted in the

laboratory.

20


1. Properly maintained biological safety cabinets are used (Class II or III) for all

manipulation of infectious materials. Biological safety cabinets (BSC) should be

located away from doors, air supplies and other heavily traveled areas. Biological

safety cabinets must be certified at least yearly.

2. Outside of a BSC, appropriate combinations of personal protective equipment are

used (e.g., special protective clothing, masks, gloves, face protection, or respirators)

in combination with physical containment devices (e.g., centrifuge safety cups,

sealed centrifuge rotors, or containment caging for animals).

3. Biological safety cabinets must be used for manipulations of cultures and clinical or

environmental materials that may be a source of infectious aerosols. The aerosol

challenge of experimental animals; harvesting of tissues or fluids from infected

animals, and embryonated eggs, and necropsy of infected animals also require the

use of BSCs.

4. Face protection (goggles and mask, or face shield) is worn for manipulations of

infectious materials outside of a biological safety cabinet.

5. Respiratory protection is worn when aerosols cannot be safety contained (e.g.,

outside of a biological safety cabinet), and in rooms containing infected animals.

6. Protective laboratory clothing such as solid-front or wrap-around gowns, scrub

suits, or coveralls must be worn in, and not worn outside, the laboratory. Reusable

laboratory clothing is to be decontaminated before being laundered. Change

protective clothing immediately if contaminated.

7. Gloves must be worn when handling infected animals and when hands may contact

infectious materials and contaminated surfaces or equipment. Disposable gloves

should be discarded when contaminated, and never washed for reuse. Always wash

hands between glove changes.


1. The laboratory is separated from areas that are open to unrestricted traffic flow

within the building. Passage through two sets of self-closing doors is the basic

requirement for entry into the laboratory from access corridors or other contiguous

areas. A clothes change room (shower optional) may be included in the

passageway.

2. Each laboratory contains a sink for hand washing. The sink is foot, elbow, or

automatically operated and is located near the laboratory exit door.

3. The interior surfaces of walls, floors, and ceilings are smooth, hard, impervious,

and water-resistant so that they can be easily cleaned and capable of disinfection.

Penetrations around these surfaces are sealed or capable of being sealed to facilitate

decontamination. Floors should be monolithic and coved to the walls.

4. Bench tops are smooth, hard and impervious to water and resistant to acids, alkalis,

organic solvents, and moderate heat.

5. Laboratory furniture is sturdy, and spaces between benches, cabinets, and

equipment are accessible for cleaning. No fabric materials are allowed.

6. Windows in the laboratory are closed and sealed.

7. A method for decontaminating all laboratory wastes is available, preferably within

the laboratory (i.e., autoclave, chemical disinfection, incineration, or other approved

decontamination method).

21

8. A ducted exhaust air ventilation system is provided. This system creates directional

airflow that draws air from “clean” areas into the laboratory toward “contaminated”

areas. The exhaust air is not recirculated to any other area of the building, and,

depending on risk assessment, discharged to the outside through a HEPA filtration

system. The outside exhaust must be dispersed away from occupied areas and air

intakes. Laboratory personnel must verify that the direction of the airflow (into the

laboratory) is proper. Visual monitoring devices, such as digital readout or

magnehelic gauges are recommended.

9. The High Efficiency Particulate Air (HEPA)-filtered exhaust air from Class II or

Class III biological safety cabinets is discharged directly to the outside or through

the building exhaust system. If the HEPA-filtered exhaust air from Class II Type A

biological safety cabinets is to be discharged to the outside through the building

exhaust air system, it is connected to this system in a manner (e.g., thimble unit

connection) that avoids any interference with the air balance of the cabinets or

building exhaust system. Exhaust air from Class II type A biological safety cabinets

may be exhausted into the laboratory if the cabinets are tested and certified at least

every twelve months. Class III biological safety cabinets must be hard ducted to the

building exhaust system.

10. Continuous flow centrifuges or other equipment that may produce aerosols are

contained in devices that exhaust air through HEPA filters before discharge into the

laboratory.

11. Vacuum lines are protected with liquid disinfectant traps or HEPA filters, or their

equivalent, which are routinely maintained and replaced as needed.

12. An eyewash/safety shower is readily available within the BSL-3 suite.

13. Biosafety Level 3 facility design and operational procedures must be documented.

The facility must be tested for verification that the design and operational

parameters have been met prior to operation. Facilities should be re-

verified/certified at least annually (HEPAs and air systems).

14. Additional environmental protection (e.g., personnel showers, containment of other

piped services and the provision for effluent decontamination) should be added

when recommended by the agent summary statement, as determined by risk

assessment, the site conditions or other applicable federal, state, or local

regulations.

15. Level 3 laboratories and animal areas are audited by The EH&S Office of

Biological Safety quarterly.

16. Openings such as around ducts, spaces between doors and frames and other

penetrations must be capable of being sealed to facilitate decontamination.

1.2.4 Animal Biosafety Level 1 (ABSL-1)

Animal Biosafety Level 1 (ABSL-1) is suitable for

work involving well characterized agents that are

not known to cause disease in healthy adult

humans, and that are of minimal potential hazard

to laboratory personnel and the environment.

22

A. Standard Practices

1. The animal facility director establishes policies, procedures, and protocols for

emergency situations. Each project is subject to pre-approval by the Institutional

Animal Care and Use Committee (IACUC) and the Institutional Biosafety

Committee (IBC). Any special practices are approved at this time.

2. Only those persons required for program or support purposes are authorized to

enter the facility. Before entering, persons are advised of the potential biohazards

and are instructed on the appropriate safeguards.

3. An appropriate medical surveillance program is in place.

4. A safety manual is prepared or adopted. Personnel are advised of special hazards,

and are required to read and follow instructions on practices and procedures.

5. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing

food for human use should only be done in designated areas and are not permitted

in animal or procedure rooms.

6. All procedures are carefully performed to minimize the creation of aerosols or

splatters.

7. Work surfaces are decontaminated after use or after any spill of viable materials.

8. All wastes from the animal room (including animal tissues, carcasses, and

contaminated bedding) are transported from the animal room in leak-proof, covered

containers for appropriate disposal in compliance with applicable institutional or

local requirements. As a means of incineration is not available on the NAU

Campus, all animal wastes must be packaged and shrouded and delivered to the

Vivarium Manager at the Biological Sciences Annex.


10. Personnel wash their hands after handling cultures and animals, after removing

gloves, and before leaving the animal facility.

11. A biohazard sign must be posted on the entrance to the animal room whenever

infectious agents are present. The hazard warning sign identifies the infectious

agent(s) in use, lists the name and telephone number of the responsible person(s),

and indicates the special requirements for entering the animal room (e.g., the need

for immunizations and respirators).


B. Special Practices: None

C. Safety Equipment (Primary Barriers):

23

1. The wearing of laboratory coats, gowns, and/or uniforms in the facility is

recommended. Laboratory coats remain in the animal room. Gowns and uniforms

are not worn outside the facility.

2. Persons having contact with non-human primates should assess their risk of mucous

membrane exposure and wear appropriate eye and face protection.

D. Facilities (Secondary Barriers)

1. The animal facility is separated from areas that are open to unrestricted personnel

traffic within the building.

2. External facility doors are self-closing and self-locking. Doors to animal rooms

open inward, are self-closing, and are kept closed when experimental animals are

present. Cubicle room inner doors may open outward or be horizontal or vertical

sliding.

3. The animal facility is designed, constructed, and maintained to facilitate cleaning

and housekeeping. The interior surfaces (walls, floors, and ceilings) are water

resistant.

4. Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, are

arranged to minimize horizontal surface areas.

5. Windows are not recommended. Any windows must be resistant to breakage.

Where possible, windows should be sealed. If the animal facility has windows that

open, they are fitted with fly screens.

6. If floor drains are provided, the traps are always filled with water and/or an

appropriate disinfectant.

7. Ventilation should be provided in accordance with the Guide for Care and Use of

Laboratory Animals, latest edition. No recirculation of exhaust air should occur. It

is recommended that animal rooms maintain negative pressure compared to

adjoining hallways.

8. The facility has a hand washing sink.

9. Cages are washed manually or in a cage washer. The mechanical cage washer

should have a final rinse temperature of at least 180ºF.

10. Illumination is adequate for all activities, avoiding reflections and glare that could

impede vision.


Animal Biosafety Level 2 involves practices for work with those

agents associated with human disease. It addresses hazards from

ingestion as well as from percutaneous and mucous membrane

exposure. ABSL-2 builds upon the practices, procedures,

containment equipment, and facility requirements of ABSL-1.

24


1. Aside from the standard policies, procedures, and protocols for emergency

situations established by the facility director, appropriate special policies and

procedures should be developed as needed and approved by the Institutional


Committee (IBC).

2. Access to the animal room is limited to the fewest number of individuals possible.

Personnel who must enter the room for program or service purposes when work is

in progress are advised of the potential hazard.

3. An appropriate medical surveillance program is in place. All personnel receive

appropriate immunizations or tests for the agents handled or potentially present

(e.g., hepatitis B vaccine, TB skin testing). When appropriate, a serum surveillance

system should be implemented.


hazards, and are required to read and follow instructions on practices and

procedures.


food for human use should only be done in designated areas and are not permitted



splatters.

7. Equipment and work surfaces in the room are routinely decontaminated with an

effective disinfectant after work with the infectious agent, and especially after overt

spills, splashes, or other contamination by infectious materials.

8. All infectious samples are collected, labeled, transported, and processed in a

manner that contains and prevents transmission of the agent(s). All wastes from the

animal room (including animal tissues, carcasses, contaminated bedding, unused

feed, sharps, and other refuse) are transported from the animal room in leak-proof,

covered containers for appropriate disposal in compliance with applicable

institutional or local requirements. The outer surface of the containers is disinfected

prior to moving the material. As a means of incineration is not available on the

NAU Campus, all animal wastes must be packaged and shrouded and delivered to

the Vivarium Manager at the Biological Sciences Annex.

9. Policies for the safe handling of sharps are instituted:

a. Needles and syringes or other sharp instruments are restricted for

use in the animal facility only when there is no alternative, such as

for parenteral injection, blood collection, or aspiration of fluids from

laboratory animals and diaphragm bottles.

b. Syringes that re-sheathe the needle, needle-less systems, and other

safe devices should be used when appropriate.

c. Plastic-ware should be substituted for glassware whenever possible.






25

and indicates the special requirements (e.g., the need for immunizations and

respirators) for entering the animal room.



1. Animal care laboratory and support personnel receive appropriate training on the

potential hazards associated with the work involved, the necessary precautions to

prevent exposures, and the exposure evaluation procedures. Personnel receive

annual updates, or additional training as necessary for procedural or policy changes.

Records of all training provided are maintained. In general, persons who may be at

increased risk of acquiring infection, or for whom infection might be unusually

hazardous, are not allowed in the animal facility unless special procedures can

eliminate the extra risk.

2. Only animals used for the experiment(s) are allowed in the room.

3. All equipment must be appropriately decontaminated prior to removal from the

room.

4. Spills and accidents which result in overt exposures to infectious materials must be

immediately reported to the facility director. Medical evaluation, surveillance, and

treatment are provided as appropriate and written records are maintained.


1. Gowns, uniforms, or laboratory coats are worn while in the animal room. The

laboratory coat is removed and left in the animal room. Gowns, uniforms, and

laboratory coats are removed before leaving the animal facility. Gloves are worn

when handling infected animals and when skin contact with infectious materials is

unavoidable.

2. Personal protective equipment is used based on risk assessment determinations.

Appropriate face/eye and respiratory protection is worn by all personnel entering

animal rooms that house nonhuman primates.

3. Biological safety cabinets, other physical containment devices, and/or personal

protective equipment (e.g., respirators, face shields) are used whenever conducting

procedures with a high potential for creating aerosols. These include necropsy of

infected animals, harvesting of tissues or fluids from infected animals or eggs, or

intranasal inoculation of animals.

4. When needed, animals are housed in primary biosafety containment equipment

appropriate for the animal species. Filter top cages are always handled in properly

designed and operating animal bio-containment cabinets recommended for rodents.




2. Access to the facility is limited by secure locked doors. External doors are self-

closing and self-locking. Doors to animal rooms open inward, are self-closing, and

are kept closed when experimental animals are present. Cubicle room inner doors

may open outward or be horizontal or vertical sliding.

26


and housekeeping. The interior surfaces (walls, floors, and ceilings) are smooth,

hard and impervious.



5. Any windows must be resistant to breakage. Where possible, windows should be

sealed. If the animal facility has windows that open, they are fitted with fly screens.

6. If floor drains are provided, the traps are always filled with an appropriate

disinfectant.

7. Exhaust air is discharged to the outside without being recirculated to other rooms.

Ventilation should be provided in accordance with criteria from Guide for Care and

Use of Laboratory Animals, latest edition. The direction of airflow in the animal

facility is inward; animal rooms should maintain negative pressure compared to

adjoining hallways.

8. Cages are washed manually or in an appropriate cage washer. The mechanical cage

washer should have a final rinse temperature of at least 180°F.

9. An autoclave is available in the animal facility to decontaminate infectious waste.

10. A hand washing sink is in the animal room where infected animals are housed, as

well as elsewhere in the facility.


impede vision.


Animal Biosafety Level 3 involves practices suitable for work with animals infected with

indigenous or exotic agents that present the potential of aerosol transmission and of causing serious

or potentially lethal disease. ABSL-3 builds upon the standard practices, procedures, containment

equipment, and facility requirements of ABSL-2.


1. Aside from the standard policies, procedures, and protocols for emergency

situations established by the facility director, appropriate special policies and

procedures should be developed as needed and approved by the Institutional


Committee (IBC).

2. The laboratory or animal facility director limits access to the animal room to the

fewest number of individuals possible. Personnel who must enter the room for

program or service purposes when work is in progress are advised of the potential

hazard(s).

3. An appropriate medical surveillance program is in place. All personnel are given

the opportunity to receive appropriate immunizations or tests for the agents handled

or potentially present (e.g., hepatitis B vaccine, TB skin testing). When appropriate,

a serum surveillance system should be implemented. In general, persons who may

be at increased risk of acquiring infection, or for whom infection might have serious

consequences, are not allowed in the animal facility unless special procedures can

27

eliminate the extra risk. Assessment should be made by the occupational health

physician.


hazards, and are required to read and follow instructions on practices and

procedures.


food for human use should be done only in designated areas and are not permitted



splatters.

7. Equipment and work surfaces in the room are routinely decontaminated with an

effective disinfectant after work with the infectious agent, and especially after overt

spills, splashes, or other contamination by infectious materials.

8. All wastes from the animal room (including animal tissues, carcasses, contaminated

bedding, unused feed, sharps, and other refuse animal tissues) are transported from

the animal room in leak-proof, covered containers for appropriate disposal in

compliance with applicable institutional or local requirements. The outer surface of

the containers is disinfected prior to moving the material. As a means of

incineration is not available on the NAU Campus, all animal wastes must be

packaged, shrouded and delivered to the Vivarium Manager at the Biological

Sciences Annex. (see Special Practices #3 below).

9. Policies for the safe handling of sharps are instituted:

a. Needles and syringes or other sharp instruments are restricted in the animal

facility for use only when there is no alternative, such as for parenteral

injection, blood collection, or aspiration of fluids from laboratory animals

and diaphragm bottles.

b. Syringes that re-sheathe the needle, needle-less systems, and other safety


c. Plastic ware should be substituted for glassware whenever possible.






and indicates the special requirements for entering the animal room (e.g., the need

for immunizations, respiratory protection, etc.).

12. All infectious samples are collected, labeled, transported, and processed in a

manner that contains and prevents transmission of the agent(s).

13. Laboratory and support personnel receive appropriate training on the potential

hazards associated with the work involved, the necessary precautions to prevent

exposures, and the exposure evaluation procedures. As necessary, personnel

receive updates and/or additional training on procedural or policy changes. Records

of all training provided are maintained.



28

1. Cages are autoclaved or thoroughly decontaminated before bedding is removed and

before they are cleaned and washed. Equipment must be decontaminated according

to any local, state, or federal regulations before being packaged for transport or

removal from the facility for repair or maintenance.

2. A spill procedure is developed and posted. Only personnel properly trained and

equipped to work with infectious materials are to clean up spills. Spills and

accidents that result in overt exposures to infectious materials must be immediately

reported to the facility director. Medical evaluation, surveillance, and treatment are

provided as appropriate and written records are maintained.

3. Materials not related to the experiment (e.g., plants, animals) are not permitted in

the animal room.


1. Uniforms or scrub suits are worn by personnel entering the

animal room. Coveralls or solid-front gowns should be

worn over this clothing. Front-button laboratory coats are

unsuitable. The gown must be removed and left in the

animal room. Before leaving the animal facility, scrub suits

and uniforms are removed and appropriately contained and

decontaminated prior to laundering or disposal.

2. Personal protective equipment used is based on risk

assessment determinations.

a. Personal protective equipment is used for all

activities involving manipulations of infectious

material or infected animals.

b. Personnel wear gloves when handling infected

animals. Gloves are removed aseptically and

autoclaved with other animal room wastes before

disposal.

c. Appropriate face/eye and respiratory protection

(e.g., PAPR) is worn by all personnel entering animal rooms.

d. Boots, shoe covers, or other protective footwear are available and used

where indicated.

3. The risk of infectious aerosols from infected animals or their bedding can also be

reduced if animals are housed in containment caging systems, such as open cages

placed in inward flow ventilated enclosures (e.g., laminar flow cabinets), solid wall

and bottom cages covered with filter bonnets, or other equivalent primary

containment systems.

4. Biological safety cabinets and other physical containment devices are used

whenever conducting procedures with a potential for creating aerosols. These

include necropsy of infected animals, harvesting of tissues or fluids from infected

animals or eggs, or intranasal inoculation of animals. At ABSL-3, all work should

be done within a primary barrier; otherwise, respirators should be worn by all

personnel in the room.


29



2. Access to the facility is limited by a self-closing and self-locking door. This exterior

entry door may be controlled by a key lock, card key, biometrics, or proximity

reader. Entry into the animal room is via a double-door entry which includes a

change room and shower(s). An additional double-door access (air-lock) or double-

doored autoclave may be provided for movement of supplies and wastes into and

out of the facility, respectively. Doors to animal rooms open inward and are self-

closing. Doors to cubicles inside an animal room may open outward or slide

horizontally or vertically.


and housekeeping. The interior surfaces (walls, floors, and ceilings) are smooth,

hard, impervious and capable of harsh disinfection. Penetrations in floors, walls and

ceiling surfaces are sealed and openings around ducts and the spaces between doors

and frames are capable of being sealed to facilitate decontamination.

4. A hands-free or automatically operated hand washing sink is provided in each

animal room near the exit door. The sink trap is filled with an appropriate

disinfectant after each use.



6. Windows are not recommended. Any windows must be resistant to breakage and

must be sealed. Direct viewing of animals is prohibited.

7. If floor drains are provided, they are always filled with an appropriate disinfectant.

8. Ventilation should be provided in accordance with criteria from the Guide for Care

and Use of Laboratory Animals, latest edition. A ducted exhaust air ventilation

system is provided. This system creates directional airflow which draws air into the

laboratory from "clean" areas and toward "contaminated" areas. The exhaust air is

not recirculated to any other area of the building. Filtration and other treatments of

the exhaust air may not be required, but should be considered based on site

requirements, and specific agent manipulations and use conditions. The exhaust

must be dispersed away from occupied areas and air intakes, or the exhaust must be

HEPA-filtered. Personnel must verify that the direction of the airflow (into the

animal areas) is proper. It is recommended that a visual monitoring device that

indicates and confirms directional inward airflow be provided at the animal room

entry. Consideration should be given to installing an HVAC control system to

prevent sustained positive pressurization of the animal spaces. Audible alarms

should be considered to notify personnel of HVAC system failure.

9. HEPA-filtered exhaust air from a Class II type A biological safety cabinet can be

exhausted into the animal room if the cabinet is tested and certified at least

annually. When exhaust air from Class II type A safety cabinets is to be discharged

to the outside through the building exhaust air system, the cabinets must be

connected in a manner that avoids any interference with the air balance of the

cabinets or the building exhaust system (e.g., a thimble unit, or an air gap between

the cabinet exhaust and the exhaust duct). When Class III biological safety cabinets

are used, they should be directly connected (hard ducted) to the exhaust system

10. Cages are washed in a cage washer. The mechanical cage washer has a final rinse

temperature of at least 180°F.

30

11. An autoclave is available which is convenient to (preferably within) the animal

rooms where the biohazard is contained. The autoclave is utilized to decontaminate

infectious waste before moving it to other areas of the facility. A pass-through

autoclave is recommended.

12. If vacuum service (i.e., central or local) is provided, each service connection should

be fitted with liquid disinfectant traps and an in-line HEPA filter, placed as near as

practicable to each use point or service cock. Filters are installed to permit in-place

decontamination and replacement.


impede vision.

14. The completed Biosafety Level 3 facility design and operational procedures must be

documented. The facility must be tested for verification that the design and

operational parameters have been met prior to operation. Facilities should be re-

verified at least annually against these procedures as modified by operational

experience.

15. Additional environmental protection (e.g., personnel showers, HEPA filtration of

exhaust air, containment of other piped services, and the provision of effluent

decontamination) should be considered if recommended by the agent summary

statement, as determined by risk assessment of the site conditions, or other

applicable federal, state, or local regulations.

31

Table 1: Summary of Recommended Biosafety Levels for Infectious Agents

BSL Agents

Practices Safety Equipment

(Primary Barriers)

Facilities

(Secondary Barriers)

1 Not known to consistently

cause disease in healthy

adults

Standard Microbiological Practices None required Open bench top

Sink for hand washing

2 Associated with human

disease, hazard =

percutaneous injury,

ingestion, mucous

membrane exposure

BSL-1 practice plus:

• Limited access

• Biohazard warning signs

• “Sharps” precautions

• Biosafety manual defining any

needed waste decontamination

or medical surveillance policies

Primary barriers = Class I or II

BSCs or other physical containment

devices used for all manipulations of

agents that cause splashes or

aerosols of infectious materials;

PPE; laboratory coats; gloves; face

protection as needed

BSL-1 plus:

• Autoclave available

• Single pass air with no

recirculation

• Hard, smooth, impervious

floor, walls, and

countertops

3 Indigenous or exotic agents

with potential for aerosol

transmission; disease may

have serious or lethal

consequences


• Controlled access

• Decontamination of all waste

• Decontamination of lab

clothing before laundering

• Baseline serum

• Decontamination of all effluent

Primary barriers = Class I or II

BSCs or other physical containment

devices for all open manipulations of

agents; PPE; protective lab clothing;

gloves; respiratory protection as

needed

BSL-2 plus:

• Physical separation from

access corridors

• Self-closing, double-door

access

• Exhausted air not re-

circulated

• Negative airflow into

laboratory (single pass)

• No floor drains

• Hard, smooth, impervious

ceilings

4 Dangerous/exotic agents

which pose high risk of

life-threatening disease,

aerosol-transmitted lab

infections; or related agents

with unknown risk of

transmission


• Clothing change before entering

• Shower on exit

• All material decontaminated on

exit from facility

Primary barriers = All procedures

conducted in Class III BSCs or Class

I or II BSCs in combination with

full-body air-supplied, positive

pressure personnel suit

BSL-3 plus:

• Separate building or

isolated zone

• Dedicated supply and

exhaust, vacuum, and

decontamination systems

• Other requirements

outlined in the text

32

Table 2: Summary of Recommended Biosafety Levels for Activities in Which Experimentally or Naturally Infected

Vertebrate Animals Are Used

ABSL Agents

Practices Safety Equipment

(Primary Barriers)

Facilities

(Secondary Barriers)

1 Not known to

consistently cause

disease in healthy

adults

Standard animal care and management

practices, including appropriate medical

surveillance programs

As required for normal care of each

species

Standard animal facility

• No recirculation of exhaust air

• Directional air flow recommended

• Hand washing sink recommended

2 Associated with

human disease,

hazard =

percutaneous injury,

ingestion, mucous

membrane exposure

ABSL-1 practice plus:

• Limited access

• Biohazard warning signs

• “Sharps” precautions

• Decontamination of all

infectious wastes and of animal

cages prior to washing

ABSL-1 equipment plus:

• Containment equipment

appropriate for animal species;

PPE; laboratory coats, gloves,

face and respiratory protection

as needed

ABSL-1 facility plus:

• Autoclave available

• Hand washing sink available in

animal room

• Mechanical cage washer used

• Hard, smooth, impervious walls,

floors, ceilings and countertops

3 Indigenous or exotic

agents with potential

for aerosol

transmission; disease

may have serious or

lethal consequences


• Controlled access

• Decontamination of all waste

• Cages decontaminated before

bedding removed

• Disinfectant foot bath as needed

• Decontamination of all effluent


• Containment equipment for

housing animals and cage

dumping activities

• Class I or II BSCs available for

manipulative procedures (e.g.,

inoculation, necropsy) that may

create infectious aerosols. PPE:

appropriate respiratory

protection


• Physical separation from access

corridors

• Self-closing, double-door access

• Sealed penetrations

• Sealed windows

• Autoclave available in facility

4 Dangerous/exotic

agents which pose

high risk of life-

threatening disease,

aerosol-transmitted

lab infections; or

related agents with

unknown risk of

transmission


• Entrance through change room

where personal clothing is

removed and laboratory

clothing is put on; shower on

exit

• All wastes are decontaminated

before removal from facility


• Maximum containment

equipment (i.e., Class III BSC

or partial containment

equipment in combination with

full body, air-supplied positive-

pressure personnel suit) used

for all procedures and activities


• Separate building or isolated zone

• Dedicated air supply and exhaust,

vacuum and decontamination

systems

• Other requirements outlined in the

text

33

1.3 Agents List

The following agents have been listed according to the most appropriate Biological Safety Level to be

used. The lists presented on the following pages are based upon the most recent information available

in the “Risk Group Classifications for Infectious Agents” section of the American Biological Safety

Association (ABSA) Website. Every attempt to provide accurate classification for the agents listed

has been made. As this is not a complete list of infectious agents, and these classifications are subject

to change, please refer to the following website: http://www.phac-aspc.gc.ca/lab-bio/res/psds-

ftss/index-eng.php, for classification of other agents, and the most up to date information for those

listed.

Please note that Biological Safety Levels are not inherent to an agent but are performance

recommendations and should be chosen after a risk assessment is completed. A proper risk

assessment takes into account the characteristics of the agent involved, the activities to be performed,

and the environment in which the work will be completed. Therefore, certain agents may be used at

different Biological Safety Levels depending upon the circumstances. For instance, human clinical

samples from HIV-positive patients may be safely handled at BSL-2. Growth of HIV in culture

should be performed under BSL-3 containment. Biological Safety Levels may be higher or lower than

what is given below for a particular agent depending upon the circumstances of its use.

The EH&S Biological Safety Officer (BSO) and the University’s Institutional Biosafety Committee

(IBC) review all projects involving recombinant DNA, infectious disease agents, and agents of

concern to livestock and agriculture and will assist you in the risk assessment process. Once the IBC

and/or the EH&S BSO assigns a Biological Safety Level, it must be adhered to unless new

information to warrant a change, in most cases from peer-reviewed literature, is provided. The IBC

and/or BSO will review the literature and make an adjustment, if warranted.

1.3.1 Risk Group 1 (RG1) Agents

Risk Group 1 agents are not associated with disease in healthy adult humans, are of minimal potential

hazard to laboratory personnel, and of minimal potential hazard to the environment; most RG1 agents

may be handled at BSL-1. This group includes Lactobacillus spp., asporogenic Bacillus subtilis or

Bacillus licheniformis, Escherichia coli-K12 (cloning strains), Baculoviruses, and adeno-associated

virus types 1 through 4 in low concentrations (<109 IP/ml)

Those agents not listed under Biological Safety Levels 2, 3 and 4 are not automatically or implicitly

classified as BSL-1; a risk assessment must be conducted based on the known and potential properties

of the agents and their relationship to agents that are listed.


Risk Group 2 agents are associated with human disease which is rarely serious and for which

preventive or therapeutic interventions are often available. They are of moderate potential hazard to

laboratory personnel and/or the environment.

http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php

http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php

34

RG2 - Bacterial Agents Including Chlamydia

• Acinetobacter baumannii (formerly Acinetobacter calcoaceticus)

• Actinobacillus

• Actinomyces pyogenes (formerly Corynebacterium pyogenes)

• Aeromonas hydrophila

• Amycolata autotrophica

• Archanobacterium haemolyticum (formerly Corynebacterium haemolyticum)

• Arizona hinshawii - all serotypes

• Bartonella henselae, B. quintana, B. vinsonii

• Bordetella including B. pertussis

• Borrelia recurrentis, B. burgdorferi

• Burkholderia (formerly Pseudomonas species) except those listed under BSL-3

• Campylobacter coli, C. fetus, C. jejuni

• Chlamydia psittaci (non-avian strains), trachomatis, C. pneumoniae

• Clostridium haemolyticum, Cl. histolyticum, Cl. novyi, Cl. septicum, Cl. tetani

• Corynebacterium diphtheriae, C. pseudotuberculosis, C. renale

• Dermatophilus congolensis

• Edwardsiella tarda

• Erysipelothrix rhusiopathiae

• Escherichia coli - all enteropathogenic, enterotoxigenic, enteroinvasive and strains bearing K1

antigen, including E. coli O157:H7

• Haemophilus ducreyi, H. influenzae

• Helicobacter pylori

• Klebsiella

• Legionella including L. pneumophila

• Leptospira interrogans - all serotypes

• Listeria

• Moraxella

• Mycobacterium (except those listed under BSL-3) including M. avium complex, M. asiaticum, M.

bovis BCG vaccine strain, M. chelonei, M. fortuitum, M. kansasii, M. malmoense, M. marinum,

M. paratuberculosis, M. scrofulaceum, M. simiae, M. szulgai, M. ulcerans, M. xenopi

• Neisseria gonorrhoea, N. meningitidis

• Nocardia asteroides, N. brasiliensis, N. otitidiscaviarum, N. transvalensis

• Rhodococcus equi

• Salmonella including S. arizonae, S. cholerasuis, S. enteritidis, S. gallinarum-pullorum, S.

meleagridis, S. paratyphi, A, B, C, S. typhi, S. typhimurium

• Shigella including S. boydii, S. dysenteriae, type 1, S. flexneri, S. sonnei

• Sphaerophorus necrophorus

• Staphylococcus aureus

• Streptobacillus moniliformis

• Streptococcus including S. pneumoniae, S. pyogenes

• Treponema pallidum, T. carateum

• Vibrio cholerae, V. parahemolyticus, V. vulnificus

• Yersinia enterocolitica

35

RG2 - Fungal Agents

• Cryptococcus neoformans

• Dactylaria galopava (Ochroconis gallopavum)

• Epidermophyton

• Exophiala (Wangiella) dermatitidis

• Fonsecaea pedrosoi

• Microsporum

• Penicillium marneffei

• Sporothrix schenckii

• Trichophyton

RG2 - Parasitic Agents

• Ancylostoma human hookworms including A. duodenale, A. ceylanicum

• Ascaris including Ascaris lumbricoides suum

• Babesia including B. divergens, B. microti

• Brugia filaria worms including B. malayi, B. timori

• Coccidia

• Cryptosporidium including C. parvum

• Cysticercus cellulosae (hydatid cyst, larva of T. solium)

• Entamoeba histolytica

• Enterobius

• Fasciola including F. gigantica, F. hepatica

• Giardia including G. lamblia

• Heterophyes

• Hymenolepis including H. diminuta, H. nana

• Isospora

• Leishmania ethiopia, L. major, L. mexicana, L. peruvania, L. tropica

• Loa filaria worms

• Microsporidium

• Necator human hookworms including N. americanus

• Onchoerca filaria worms including, O. volvulus

• Plasmodium including simian species, P. cynomologi, P. falciparum, P. malariae, P. ovale, P.

vivax

• Sarcocystis including S. sui hominis

• Schistosoma including S. haematobium, S. intercalatum, S. japonicum, S. mansoni, S. mekongi

• Strongyloides including S. stercoralis

• Toxocara including T. canis

• Toxoplasma including T. gondii

• Trichinella spiralis

• Trypanosoma including T. brucei, T. brucei gambiense, T. brucei rhodesiense, T. cruzi

• Wuchereria bancrofti filaria worms

RG2 - Viruses

Adenoviruses, human - all types except African Swine Fever virus

Arenaviruses

36

• Lymphocytic choriomeningitis virus (non-neurotropic strains)

• Other viruses as listed by ABSA/CDC

Bunyaviruses

• Bunyamwera virus

• Rift Valley fever virus vaccine strain MP-12 only


Calciviruses

• Norovirus

• Sapovirus

Coronaviruses except SARS

Flaviviruses (Togaviruses) - Group B Arboviruses

• Yellow fever virus vaccine strain 17D


Hepatitis A, B, C, D, and E viruses

Herpesviruses - except Herpesvirus simiae (Monkey B virus), BSL-4

• Cytomegalovirus

• Epstein Barr virus

• Herpesvirus ateles

• Herpesvirus saimiri

• Herpes simplex types 1 and 2

• Herpes zoster

• Human herpesvirus types 6 and 7

Marek's disease virus

Murine cytomegalovirus

Pseudorabies virus

Orthomyxoviruses

• Influenza viruses types A, B, and C

• Other tick-borne orthomyxoviruses as listed by ABSA/CDC

Papovaviruses

• All human papilloma viruses

• Bovine papilloma virus

• Polyoma virus

• Shope papilloma virus

• Simian virus 40 (SV40)

Paramyxoviruses

• Newcastle disease virus (non-exotic, non-virulent strains)

• Measles virus

• Mumps virus

• Parainfluenza viruses types 1, 2, 3, and 4

37

• Respiratory syncytial virus

Parvoviruses

• Human parvovirus (B19)

• Canine Parvovirus (CPV)

Picornaviruses

• Coxsackie viruses types A and B

• Echoviruses - all types

• Polioviruses - all types, wild and attenuated

• Rhinoviruses - all types

Poxviruses

• Vaccinia - all types except Monkeypox virus (BSL-3) and restricted poxviruses including

Alastrim, Camelpox, Goatpox, Smallpox, and Whitepox (restricted to the CDC, Atlanta, GA)

Reoviruses - Coltivirus, human Rotavirus, and Orbivirus (Colorado tick fever virus)

Retroviruses

• Bovine leukemia virus

• Clinical samples from HIV-positive patients

• Feline immunodeficiency virus

• Feline leukemia virus

• Feline sarcoma virus

• Gibbon leukemia virus

• Mason-Pfizer monkey virus

• Mouse mammary tumor virus

• Murine leukemia virus

• Murine sarcoma virus

• Rat leukemia virus

NOTE: Containment requirements for research involving rDNA

Experiments involving recombinant DNA lend themselves the application of highly specific biological

barriers. The containment levels required for research involving recombinant DNA associated with

plants or animals is based on classification of experiments in the NIH Guidelines Section III

(“Experiments Covered by the NIH Guidelines”), which can be found at

http://osp.od.nih.gov/sites/default/files/resources/NIH_Guidelines.pdf. Consult this document for

containment requirements specific to your research.

Rhabdoviruses

• Mokola Virus

• Vertebrate and Invertebrate lyssaviruses

Togaviruses (see Alphaviruses and Flaviviruses)

• Rubivirus (rubella)

• Ross River Virus

http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm#_blank

http://osp.od.nih.gov/sites/default/files/resources/NIH_Guidelines.pdf

38


Risk Group 3 agents are associated with serious or lethal human disease, for which preventive or

therapeutic interventions may be available.

RG3 - Bacterial Agents Including Rickettsia

• Bacillus anthracis

• Bartonella

• Brucella including B. abortus, B. canis, B. suis

• Burkholderia (Pseudomonas) mallei, B. pseudomallei

• Clostridium botulinum, Cl. chauvoei

• Coxiella burnetii

• Francisella tularensis (Types A and B)

• Mycobacterium bovis (except BCG strain, BSL-2), M. tuberculosis, M. leprae

• Mycoplasma, except M. mycoides and M. agalactiae which are restricted animal pathogens

• Pasteurella multocida type B - “buffalo” and other virulent strains

• Rickettsia akari, R. australis, R. canada, R. conorii, R. prowazekii, R. rickettsii, R, siberica, R.

tsutsugamushi, R. typhi (R. mooseri)

• Yersinia pestis

RG3 - Fungal Agents

• Blastomyces dermatitidis

• Cladosporium bantianum, C. (Xylohypha) trichoides

• Coccidioides immitis (sporulating cultures; contaminated soil)

• Histoplasma capsulatum, H. capsulatum var. duboisii

• Paracoccidioides braziliensis


• Echinococcus including E. granulosis, E. multilocularis, E. vogeli

• Leishmania including L. braziliensis, L. donovani

• Naegleria fowleri

• Plasmodium falciparum

• Taenia solium

RG3 - Viruses and Prions

Alphaviruses (Togaviruses) - Group A Arboviruses

• Eastern equine encephalomyelitis virus

• Venezuelan equine encephalomyelitis vaccine strain TC-83

• Western equine encephalomyelitis virus

• Semliki Forest virus

• Venezuelan equine encephalomyelitis virus (except the vaccine strain TC-83 is BSL-2)


Arenaviruses

• Lymphocytic choriomeningitis virus (LCM) (neurotropic strains)

• Flexal

39

Bunyaviruses

• Hantaviruses including Hantaan virus

• Rift Valley fever virus


Flaviviruses (Togaviruses) - Group B Arboviruses

• Dengue virus serotypes 1, 2, 3, and 4

• Japanese encephalitis virus

• St. Louis encephalitis virus

• Yellow fever virus – wild type


Poxviruses

• Monkeypox virus

Prions

• Transmissible spongioform encephalopathies (TME) agents, Creutzfeldt-Jacob disease and kuru

agents (see BMBL for specific containment instruction)

Retroviruses

• Avian leukosis virus

• Avian sarcoma virus

• Human immunodeficiency virus (HIV) types 1 and 2

• Human T cell lymphotropic virus (HTLV) types 1 and 2

• Simian immunodeficiency virus (SIV)

Rhabdoviruses

• Rabies virus

• Vesicular stomatitis virus - laboratory adapted strains ONLY including VSV-Indiana, San Juan,

and Glasgow


Risk Group 4 agents are likely to cause serious or lethal human disease for which preventive or

therapeutic interventions are not usually available.

RG4 - Bacterial Agents

None

RG4 - Fungal Agents

None


None

40

RG4 - Viral Agents

Arenaviruses (Togaviruses) - Group A Arboviruses

• Guanarito virus

• Lassa virus

• Junin virus

• Machupo virus

• Sabia virus

Bunyaviruses (Nairovirus)

• Crimean-Congo hemorrhagic fever virus

Filoviruses

• Ebola virus

• Marburg virus

Flaviruses (Togaviruses) - Group B Arboviruses

• Tick-borne encephalitis virus complex including Absetterov, Central European encephalitis,

Hanzalova, Hypr, Kumlinge, Kyasanur Forest disease, Omsk hemorrhagic fever, and Russian

spring-summer encephalitis viruses

Herpesviruses (alpha)

• Herpesvirus simiae (Herpes B or Monkey B virus)

Paramyxiviruses

• Equine morbillivirus

Hemorrhagic fever agents and viruses as yet undefined.

2.0 INFORMATION FOR RESEARCHERS

2.1 Project Registration

Northern Arizona University Institutional Biosafety Committee

Many research projects involve work with potentially hazardous biological agents, known infectious

disease agents, or biological materials regulated by the federal or state government. Granting agencies

require that the university monitor the use of biological hazards, infectious disease agents, and

recombinant DNA in order for them to release funds to investigators. Therefore, we have developed a

registration system to ensure that all biological materials are handled properly and disposed of

appropriately. The EH&S Office of Biological Safety administers three registration programs for

research, teaching, and clinical laboratories.

The NAU Institutional Biosafety Committee (IBC) reviews all registrations. Approval by the NAU

IBC, in writing, is required before ordering or working with any agents. Please contact the EH&S

BSO for more information.

Fillable registration forms are available on the EH&S website at:

http://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Biological-Safety/

http://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Biological-Safety/

41

2.1.1 Biological Agent (BA) Registration

Use of the following materials requires that the Principal Investigator completes and submits the

biological agent registration document for approval by the EH&S Biological Safety Officer. Agents,

such as plant pathogens or exotic microorganisms, that are regulated by federal or state agencies

(CDC, HHS, DOC, USDA/APHIS, EPA, FDA, DPI, etc.) shall be registered with the EH&S Office

of Biological Safety by submission of a biological agent registration form. All permits for transport,

transfer, import or export of these regulated agents are the responsibility of the Principal Investigator.

The NAU Responsible Official (RO) will be responsible for all Select Agent permits. Please allow a

minimum of 6 weeks to obtain new federal permits.

Agents characterized at Biosafety Level 1 require registration. These will be expedited and

administratively approved barring any complications.

Agents to be used at Biosafety Level 2 (BSL-2) or Biosafety Level 3 (BSL-3):

1. All human, animal, or plant pathogens that require BSL-2 or BSL-3 containment and handling

(see previous section: “Agents List”) must be registered. Please note that BSL-4 agents may

not be stored or used at NAU.

2. Unknown human and animal pathogens must be registered. These are considered BSL-2 until

identified.

3. Cell lines or cultures that

1) have been immortalized with a virus (such as EBV or a retrovirus),

2) are known to be tumorigenic in primates (including humans), or

3) are primary human tumor cells.

These are considered BSL-2 (or higher in many cases).

4. Human blood or other tissues, when used in research, must be registered.

5. Human blood or other tissue known to be HIV positive or known to contain any human

disease causing agent may require higher containment depending upon the IBC evaluation.

2.1.2 Recombinant DNA (RD) Registration

All recombinant-DNA (rDNA) projects that involve a recombinant organism (this excludes projects

that involve DNA only, i.e., PCR products) require registration with the EH&S Office of Biological

Safety. All rDNA projects require review and approval from the Institutional Biosafety Committee

(IBC), which oversees all research projects and issues involving rDNA at NAU. Use of the following

requires that the principal investigator complete and submit an rDNA registration document.

1. All rDNA projects, including the growth of recombinant bacteria for probe isolation (plasmid

or phage preparations) require registration. Projects must be registered regardless of where the

material came from or who originally constructed it.

2. Projects that are exempt from the NIH Guidelines must also be registered.

3. The development or use of transgenic animals and plants requires registration.

rDNA projects are performed at BSL-1, BSL-2, BSL-3 or the corresponding levels for whole plant

(BSL-1P, BSL-2P, BSL-3P) or whole animal (BSL-1N, BSL-2N, BSL-3N) work. The EH&S

Biological Safety Officer, in conjunction with the IBC, will make the final determination.

42

2.1.3 Acute Toxins (AT) Registration

The use and storage of toxins with a mammalian LD50 of < 100 g/kg require registration. For a

partial list, see the Toxins Table that follows. Acute toxins may only be ordered following written

approval by the BSO or IBC.

2.1.4 Project Amendments

Changes to an existing registration can be done on an amendment unless said changes result in a

dramatic change to the overall project or the containment level.

Typical changes include:

• Addition or deletion of new personnel

• Addition or deletion of new agents (same containment level)

• Minor modifications to the protocol or procedures (final decision to be made by the IBC)

43

Table 3: Toxin Table

Toxins with a mammalian LD50 of < 100 g/kg must be registered with the EH&S Office of

Biological Safety. Therefore, use of the following toxins requires registration. If a toxin is not on the

list, it still may require registration, depending upon the LD50. For more information, please contact

the EH&S Office of Biological Safety.

Toxicity

LD50 (g/kg)*

Abrin 0.7

Aerolysin 7.0

Botulinin toxin A 0.0012

Botulinin toxin B 0.0012

Botulinin toxin C1 0.0011

Botulinin toxin C2 0.0012

Botulinin toxin D 0.0004

Botulinin toxin E 0.0011

Botulinin toxin F 0.0025

-bungarotoxin 14.0

Caeruleotoxin 53

Cereolysin 40-80

Cholera toxin 250

Clostridium difficile enterotoxin A 0.5

Clostridium difficile cytotoxin B 220

Clostridium perfringens lecithinase 3

Clostridium perfringens kappa toxin 1500

Clostridium perfringens perfringolysin O 13-16

Clostridium perfringens enterotoxin 81

Clostridium perfringens beta toxin 400

Clostridium perfringens delta toxin 5

Clostridium perfringens epsilon toxin 0.1

Conotoxin 12-30

Crotoxin 82

Diphtheria toxin 0.1

Listeriolysin 3-12

Leucocidin 50

Modeccin 1-10

Nematocyst toxins 33-70

Notexin 25

Pertussis toxin 15

44

Toxicity

LD50 (g/kg)*

Pneumolysin 1.5

Pseudomonas aeruginosa toxin A 3

Ricin 2.7

Saxitoxin 8

Shiga toxin 0.250

Shigella dysenteriae neurotoxin 1.3

Streptolysin O 8

Staphylococcus enterotoxin B 25

Staphylococcus enterotoxin F 2-10

Streptolysin S 25

Taipoxin 2

Tetanus toxin 0.001

Tetrodotoxin 8

Viscumin 2.4-80

Volkensin 1.4

Yersinia pestis murine toxin 10

*Please note that the LD50 values are from a number of sources (see below). For specifics on route of

application (i.v., i.p., s.c.), animal used, and variations on the listed toxins, please go to the references

listed below.

Reference:

1. Gill, D. Michael; 1982; Bacterial toxins: a table of lethal amounts; Microbiological Reviews;

46: 86-94

2. Stirpe, F.; Luigi Barbieri; Maria Giulia Battelli, Marco Soria and Douglas A. Lappi; 1992;

Ribosome-inactivating proteins from plants: present status and future prospects;

Biotechnology; 10: 405-412 3. Registry of toxic effects of chemical substances (RTECS): comprehensive guide to the

RTECS. 1997. Doris V. Sweet, ed., U.S. Dept. of Health and Human Services, Public Health

Service, Centers for Disease Control and Prevention, National Institute for Occupational

Safety and Health; Cincinnati, Ohio

2.1.5 Select Agents

The following lists of agents and toxins are classified by the Federal government as Select Agents.

Any possession, use, transfer or shipment of these materials is strictly controlled by regulation.

Researchers considering work with any of these materials must first contact the NAU Responsible

Official for the approvals, permits, clearances and other necessary paperwork. Be aware that

government clearance can take as many as 6 months to complete and should be sought far in advance

of any project’s proposed start date.

Failure to comply with these Federal Regulations is punishable by both fines and imprisonment.

45

HHS and USDA Select Agents and Toxins

7CFR Part 331, 9 CFR Part 121, and 42 CFR Part 73

HHS SELECT AGENTS AND TOXINS

Abrin

Bacillus cereus Biovar anthracis*

Botulinum neurotoxins*

Botulinum neurotoxin producing species

of Clostridium*

Conotoxins (Short, paralytic alpha conotoxins

containing the following amino acid sequence

X1CCX2PACGX3X4X5X6CX7)1

Coxiella burnetii

Crimean-Congo haemorrhagic fever virus

Diacetoxyscirpenol

Eastern Equine Encephalitis virus3

Ebola virus*

Francisella tularensis*

Lassa fever virus

Lujo virus

Marburg virus*

Monkeypox virus3

Reconstructed replication competent forms of the

1918 pandemic influenza virus containing any

portion of the coding regions of all eight gene

segments (Reconstructed 1918 Influenza virus)

Ricin

Rickettsia prowazekii

SARS-associated coronavirus (SARS-CoV)

Saxitoxin

South American Haemorrhagic Fever viruses:

Chapare

Guanarito

Junin

Machupo

Sabia

Staphylococcal enterotoxins A,B,C,D,E subtypes

T-2 toxin

Tetrodotoxin

Tick-borne encephalitis complex (flavi) viruses:

Far Eastern subtype

Siberian subtype

Kyasanur Forest disease virus

Omsk hemorrhagic fever virus

Variola major virus (Smallpox virus)*

Variola minor virus (Alastrim)*

Yersinia pestis*

OVERLAP SELECT AGENTS AND

TOXINS

Bacillus anthracis*

Bacillus anthracis Pasteur strain

Brucella abortus

Brucella melitensis

Brucella suis

Burkholderia mallei*

Burkholderia pseudomallei*

Hendra virus

Nipah virus

Rift Valley fever virus

Venezuelan equine encephalitis virus3

USDA SELECT AGENTS AND TOXINS

African horse sickness virus

African swine fever virus

Avian influenza virus3

Classical swine fever virus

Foot-and-mouth disease virus*

Goat pox virus

Lumpy skin disease virus

Mycoplasma capricolum3

Mycoplasma mycoides3

Newcastle disease virus2,3

Peste des petits ruminants virus

Rinderpest virus*

Sheep pox virus

Swine vesicular disease virus

USDA PLANT PROTECTION AND

QUARANTINE (PPQ)

SELECT AGENTS AND TOXINS

Peronosclerospora philippinensis

(Peronosclerospora sacchari)

Phoma glycinicola (formerly Pyrenochaeta

glycines)

Ralstonia solanacearum

Rathayibacter toxicus

Sclerophthora rayssiae

Synchytrium endobioticum

Xanthomonas oryzae

46

*Denotes Tier 1 Select Agent

1 C = Cysteine residues are all present as disulfides, with the 1st and 3rd Cysteine, and the 2nd and

4th Cysteine forming specific disulfide bridges; The consensus sequence includes known toxins α-MI

and α-GI (shown above) as well as α-GIA, Ac1.1a, α-CnIA, α-CnIB; X1 = any amino acid(s) or Des-

X; X2 = Asparagine or Histidine; P = Proline; A = Alanine; G = Glycine; X3 = Arginine or Lysine;

X4 = Asparagine, Histidine, Lysine, Arginine, Tyrosine, Phenylalanine or Tryptophan; X5 = Tyrosine,

Phenylalanine, or Tryptophan; X6 = Serine, Threonine, Glutamate, Aspartate, Glutamine, or

Asparagine; X7 = Any amino acid(s) or Des X and; “Des X” = “an amino acid does not have to be

present at this position.” For example if a peptide sequence were XCCHPA then the related peptide

CCHPA would be designated as Des-X. 2 A virulent Newcastle disease virus (avian paramyxovirus serotype 1) has an intracerebral

pathogenicity index in day-old chicks (Gallus gallus) of 0.7 or greater or has an amino acid sequence

at the fusion (F) protein cleavage site that is consistent with virulent strains of Newcastle disease

virus. A failure to detect a cleavage site that is consistent with virulent strains does not confirm the

absence of a virulent virus. 3 Select agents that meet any of the following criteria are excluded from the requirements of this part:

Any low pathogenic strains of avian influenza virus, South American genotype of eastern equine

encephalitis virus , west African clade of Monkeypox viruses, any strain of Newcastle disease virus

which does not meet the criteria for virulent Newcastle disease virus, all subspecies Mycoplasma

capricolum except subspecies capripneumoniae (contagious caprine pleuropneumonia), all subspecies

Mycoplasma mycoides except subspecies mycoides small colony (Mmm SC) (contagious bovine

pleuropneumonia), and any subtypes of Venezuelan equine encephalitis virus except for Subtypes

IAB or IC, provided that the individual or entity can verify that the agent is within the exclusion

category. 9/10/13

2.2 Minors in Research Laboratories or Animal Facilities

Unless enrolled as a Northern Arizona University student, minors are not allowed to work or conduct

research in Northern Arizona University research laboratories, greenhouses or animal facilities except

as identified specifically below. In addition, minors are prohibited from operating farm machinery or

state vehicles and from working in machine shops. All applicable forms can be found on the EH&S

Forms webpage at http://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Forms/.

1. All Minors are prohibited from working or conducting research in the following:

a. Any laboratory or facility designated as BSL-3, ABSL-3 or higher for recombinant or

infectious organisms.

b. Any laboratory where Select Agents or explosives are used or stored.

c. Any Animal Care Services (ACS) housing or procedure area/lab/facility.

2. Minors are prohibited from working with any of the following materials.

a. Radioactive materials or radiation (X-rays)

b. Acute Toxins

3. Minors are allowed to work or conduct research in laboratories (not listed in #1 or #2 above) if

the following requirements are met in full:

a. Northern Arizona University EH&S Office of Biological Safety Policy titled; Minors

In Research Laboratories Or Animal Facilities (as been read and understood.

b. A Minor’s Research Proposal Registration Form is submitted to and approved by

http://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Forms/

47

the Northern Arizona University Institutional Biosafety Committee which includes:

i. A description of the project and all materials

ii. A parental consent form

iii. A sponsor consent form

c. Hazard specific safety training is completed by the Principal Investigator/Sponsor with

the minor as approved by EH&S BSO.

d. Personal protective equipment, specific to the hazard, is provided to the minor with

instructions for use and disposal.

e. The minor is supervised at all times while in the laboratory.

f. Hours of work comply with Federal Regulation 29 CFR 570.35.

The laboratory is in full compliance with all applicable Northern Arizona University

safety programs and regulations.

2.3 Principle Investigator’s Responsibilities During the Conduct of Research

The Principal Investigator shall:

1. Supervise the safety performance of the laboratory staff to ensure that the required safety

practices and techniques are employed.

2. Report any significant incident, violation of the NIH Guidelines, or any significant, research-

related accidents and illnesses immediately by contacting the Biological Safety Officer.

Examples of incidents and violations include:

a. Overt exposures (exposures that result in direct personnel exposure to biohazards

such as injection, spills, splashes or aerosol inhalation)

b. Potential exposures (exposures that have a high risk of exposing personnel to

biohazards such as spills, containment failure while working with the agent or

equipment failure that may produce aerosols)

c. Any exposure (overt or potential) in a BSL-3 laboratory

d. Overt exposure in BSL-1 or BSL-2 laboratories

e. Any illness that may be caused by the agents used in the laboratory

f. Incidents involving the improper disposal of biohazards

3. Ensure the integrity of the physical containment (e.g., biological safety cabinets) and the

biological containment (e.g., purity and genotypic and phenotypic characteristics) and correct

procedures or conditions that might result in release of or exposure to biohazards.

4. Limit or restrict access to the laboratory when work with biohazards is in progress; this

includes making a determination of who may be at increased risk.

5. Establish policies and procedures to limit access exclusively to those individuals who have

been advised of the potential hazards and meet specific entry requirements.

6. Ensure that laboratory personnel are offered, at no cost, appropriate immunizations or tests for

the infectious agents handled or potentially present in the laboratory (e.g., hepatitis B vaccine,

tuberculosis skin testing).

2.3.1 Reportable Incidents and Violations

Incidents/problems involving biohazards must be immediately reported to the Office of Biological

Safety. Examples of reportable significant incidents include but are not limited to:

48

1. Any overt exposure, such as a needle stick, splash, and contamination due to equipment

failure.

2. Any potential exposure in a BSL-3 facility.

A significant event may also occur from a containment breach, which may be subsequently

determined to pose either an overt or potential exposure to individuals or the environment. It should be

noted that waste from recombinant or synthetic nucleic acid research is also considered biohazardous

and incidents involving improper disposal of recombinant or synthetic nucleic acids must also be

reported. Questions regarding reportable incidents should be directed to the Office of Biological

Safety.

Failure by research personnel to follow federal and institutional regulations, guidelines, policies and/or

procedures may also require reporting to the appropriate institutional, local, state and/or federal

agencies. Violations may include but are not limited to conduct of new or ongoing research without

appropriate federal or institutional registration, review, approval or oversight.

2.3.2 Institutional Reporting Responsibilities

The Institutional Biosafety Committee is required, by the NIH Guidelines, to report to the appropriate

University official and to the NIH/OBA within thirty days any significant incidents, violations of the

NIH Guidelines, or any significant findings of research-related accidents and illnesses. The IBC will

be responsible to determine what actions, if any, are necessary. For example, the IBC may determine

the need to make changes to the frequency of laboratory inspections or biosafety containment level of

the research, based on results of the incident.

Other IBC reporting requirements (to OBA and other agencies) include but are not limited to:

1. Research involving biohazards conducted without prior IBC approval

2. Lax security, unsafe procedures used in a laboratory setting, improper disposal of recombinant

or synthetic nucleic acid waste

3. Changes to research risk that have been initiated without prior approval by IBC

Some incidents must be reported to OBA on an expedited basis. Spills or accidents in BSL-2

laboratories involving recombinant or synthetic nucleic acids that result in an overt exposure must be

immediately reported to OBA. In addition, spills or accidents involving recombinant or synthetic

nucleic acids occurring in high containment (BSL-3 or higher) laboratories resulting in an overt or

potential exposure must be immediately reported to OBA. The IBC working through the IBC Chair

and the BSO will report to the Institutional Official, who, in turn will oversee the report to OBA, any

of the above-described incidents.

Institutional violations that will also be reported to the appropriate College or department head may

include but are not limited to:

1. Lapses in disclosure approval

2. Failure to comply with institutional and federal regulations, guidelines, and policies

3. Unsafe work practices

2.4 Biological Waste Disposal Policy

This policy is intended to provide guidance and insure compliance with the NIH/CDC guidelines, the

State of Arizona Administrative Code, and restrictions of the Coconino County Landfill.

49

2.4.1 Categories

Infectious/potentially infectious/R-DNA

a) human pathogens

b) animal pathogens

c) plant pathogens

d) recombinant DNA

e) human and primate blood, blood products and other body fluids

f) human and primate tissue

g) any material containing or contaminated with any of the above (test tubes, needles*, syringes,

tubing, culture dishes, flasks, etc.)

This waste must be inactivated prior to disposal. The preferred method is steam sterilization

(autoclaving), although chemical inactivation or incineration may be appropriate in some cases. Per

NAU policy, even autoclaved or otherwise inactivated wastes must be disposed of as biohazardous

waste. Storage of non-inactivated waste is restricted to within the generating laboratory. The material

may not be stored longer than 24 hours prior to inactivation.

Non-infectious waste

This category includes waste that is not contaminated with any of the biological materials listed in

category 1. It includes solid waste and sharps generated in clinical or laboratory settings. Sterile or

unopened biomedical materials that require disposal are also considered biological waste, and must be

treated as such.

IV packs test tubes Petri dishes

needles* razor blades* tissue culture flasks

syringes culture dishes

scalpels* flasks

broken glass and plastic ware** pipettes

This material does not require sterilization prior to disposal.

*must be packaged in plastic sharps boxes.

**must be within a box or other puncture proof container before adding to waste.

Mixed radioactive/biohazardous waste

The biohazardous component of mixed radioactive/biohazardous waste shall be inactivated prior to its

release to Radiation Safety for disposal as radioactive waste. Steam sterilization or chemical

inactivation shall be employed as above. Although some radioactive materials can be autoclaved

safely, please check with the EH&S Radiation Safety Officer regarding the best method to employ

with any given radionuclide.

Mixed chemical/biohazardous waste

The biohazardous component of mixed chemical/biohazardous waste shall be inactivated prior to its

release for chemical disposal. Precautions should be taken to prevent the generation and release of

toxic chemicals during the inactivation process. In general, autoclaving is not recommended because

50

flammable or reactive compounds should not be autoclaved due to the explosion hazard. Please check

with the NAU Chemical Hygiene Officer for guidance regarding particular chemicals.

Animal carcasses and materials

As there are no tissue digesters or incinerators present on the NAU

Campus, all animal materials must be packaged, shrouded and delivered to

the Vivarium Manager. No animal carcasses or material shall be disposed

of as regular trash or through the biomedical waste receptacle.

Human remains

Please contact the State Anatomical Review Board for information

regarding the final disposition of human remains and body parts.

2.4.2 Packaging

Biohazard bags

These are used for the initial collection of certain biological wastes. All biohazard bags must meet

impact resistance (165 grams), tearing resistance (480 grams), and heavy metal concentration (<100

PPM total of lead, mercury, chromium and cadmium) requirements. Written documentation (a test

report) from the manufacturer regarding these requirements must be on file. Individual departments

are responsible for obtaining the appropriate biohazard bags. These bags must be placed in bulk

biohazard containers as described within this section.

Sharps

Needles, scalpels and razor blades must be containerized in red plastic sharps

containers. NAU departments are responsible for obtaining their own sharps

containers. All other sharps (broken glass and plastic ware, pipettes, etc.) must be

containerized in puncture-resistant cardboard boxes (see #3, below).

Containers

All biological waste must be containerized in rigid, leak proof, puncture resistant boxes or plastic

tubs/barrels as the terminal receptacle. NAU Departments are responsible for obtaining the

appropriate containers from the EH&S Hazardous Waste Coordinator.

2.4.3 Labeling

Only manufactured containers with the preprinted universal biohazard symbol and the words

“biomedical”, “biohazardous”, or “infectious” shall be used.

2.4.4 Transport

The transport of biohazardous waste outside of the laboratory (i.e., to an autoclave or incinerator)

must be in a closed, leak-proof container that is labeled “biohazard”. Only trained personnel may

transport biohazardous waste. Labeling may be accomplished by use of a red biohazard bag with the

51

universal biohazard symbol. Only biohazard bags and red plastic sharps containers may be used to

transport biological waste to the biohazard waste receptacle. Waste receptacle personnel are

instructed not to accept any other type of containers.

Transportation of red-bagged waste must be in closed, leak-proof containers, properly labeled as

“biohazards.” Movement of regulated/biological waste through public corridors, along carpeted

hallways, and on public elevators should be avoided whenever possible. Any leakage/spills from these

containers must be immediately reported to the NAU Office of Biological Safety. Signs must be

displayed to prevent tracking of the spills to other areas.

2.4.5 Training

All employees who handle biological waste shall be trained regarding its proper handling before they

are allowed to handle biological waste.

Training may be accomplished through the NAU Basic Biosafety Training Program (administered by

NAU EH&S Office of Biological Safety), informally in the lab setting, or through formal training

programs set up by individual departments or divisions. For assistance, please call the NAU

Biological Safety Officer or Hazardous Waste Coordinator.

According to Arizona Statute (Ch. 64E-16 F.A.C.), records of the training session shall be maintained

for each employee, along with an outline of the training program. Training records shall be retained

for a period of three (3) years.

2.5 Biological Waste Disposal Containers

The following waste disposal containers for biohazardous or biomedical waste are to be used for

teaching and research purposes at Northern Arizona University:

ALL BIOHAZARDOUS WASTE MUST BE TREATED BEFORE DISPOSAL.

2.5.1 Bulk Biohazard Receptacles

The EH&S currently provides departments who generate biohazardous and biomedical waste with

large receptacles for the disposal of full biohazard bags and sharps containers. Individual departments

can contact the EH&S Hazardous Waste Coordinator for pickup when those containers are full

(https://nau.edu/research/compliance/environmental-health-and-safety/hazardous-waste-

management/).

PLEASE NOTE: PER NAU POLICY, EVEN AUTOCLAVED WASTES MUST BE TREATED

AS BIOHAZARDOUS WASTES.

Disposal of biomedical waste in the regular trash or dumpster is prohibited. The Coconino County

Landfill does not accept any items that resemble “hazardous materials” and will reject the entire load.

In some cases, the individual principal investigator has been made responsible for the costs incurred

for sorting and disposal of improperly disposed waste.

https://nau.edu/research/compliance/environmental-health-and-safety/hazardous-waste-management/

https://nau.edu/research/compliance/environmental-health-and-safety/hazardous-waste-management/

52

2.5.2 Sharps Boxes

NAU is subject to the requirements of Arizona Administrative Code Rule

18-13-1419, Medical Sharps. This regulation requires that a generator who

treats biohazardous medical waste on site shall place medical sharps in a

sharps container after rendering them incapable of creating a stick hazard

by using an encapsulation agent or any other process that prevents a stick

hazard.

Red plastic sharps boxes are used for disposal of needles, razor blades,

scalpels, and small Pasteur pipettes. Sharps boxes that contain infectious

materials must be inactivated by autoclaving the closed box. Only trained

staff/research staff shall handle the full sharps boxes. Full sharps containers

can be disposed of in the bulk biohazard receptacles described above.

2.5.3 Biohazard Bags

Red plastic biohazard bags shall be used for collection of biohazardous tissue culture items, petri

dishes, and other non-sharp items. The biohazardous waste items must be inactivated by autoclaving

in biohazard autoclave bags or treatment with bleach within 24 hours of accumulation. After

treatment, the bags shall be placed in the bulk containers described above. They may only be

transported to the waste receptacle by trained laboratory personnel or by trained staff. Biohazard bags

should never be placed in an area with unrestricted access. Double bagging may be required for waste

bags containing significant liquid volumes, however, sufficient absorbent material must also be added

to any bag with liquid waste to further help prevent leakage. Full, untreated biohazard bags shall be

stored only in the lab where accumulation occurred. Full biohazard bags must be treated within 24

hours. As previously noted, even autoclaved wastes must be disposed of as biohazardous waste.

2.6 Autoclave Use and Testing

Standard operating procedures for autoclave operations and verification can be found in the Northern Arizona University Autoclave Manual on the EH&S website at https://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Standard-Operating-Procedures/. For additional questions, please contact the Office of Biological Safety.

2.7 Disinfectants

“Disinfectant” refers to an agent that is applied to treat (usually) inanimate objects to eliminate some,

but not necessarily all, microorganisms on the object to make it safer for handling. In contrast, the

term “sterilant” refers to an agent that renders items free of all microorganisms. The two are not the

same and should not be confused.

Disinfectants are used in laboratory and chemical settings to 1) treat a surface or an item before or

after routine use, or 2) treat a surface or an item after a spill or “contaminating event”.

Because disinfectants are antimicrobial, they may, by their nature, also have a toxic effect to the user.

Therefore, Material Safety Data Sheets and other manufacturer’s product information should be

available and thoroughly reviewed before using these products.

https://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Standard-Operating-Procedures/

https://nau.edu/Research/Compliance/Environmental-Health-and-Safety/Standard-Operating-Procedures/

53

There are many different types and formulations of disinfectants. The researcher or clinician should

ensure that the proper product, one that is effective against the specific microorganism being studied,

is used.

The FDA regulates those products that are marketed as sterilants or sanitizing agents for medical

devices, and has published a list of FDA-Cleared Sterilants and Disinfectants at

https://www.fda.gov/medicaldevices/deviceregulationandguidance/reprocessingofreusablemedicaldevi

ces/ucm437347.htm.

The EPA regulates moderate and low level disinfectants under their pesticide regulations. They have

published a list of Selected EPA-registered Disinfectants at https://www.epa.gov/pesticide-

registration/selected-epa-registered-disinfectants.

Please contact the Office of Biological Safety for information about any of these lists. Be aware that

most disinfectants assume pre-cleaning to remove gross organic/protein prior to use.

Whenever a disinfectant or sterilant is used, proper safety precautions must be followed. Appropriate

personal protective equipment (PPE) (e.g., gloves, safety goggles, aprons) must be worn. In addition,

these compounds must be used in well-ventilated areas.

Following is a discussion of general categories of disinfectants. Please note that there are several

different products and different formulations in each category.

2.7.1 Liquids

Alcohols

The most commonly used alcohols, ethanol and isopropanol, are most effective at concentrations of

70% in water. Both higher and lower concentrations are less effective. Alcohols are active against

vegetative bacteria, fungi, and lipid viruses but not against spores. They are only moderately effective

against nonlipid viruses. Alcohols are difficult to use as contact disinfectants because they evaporate

rapidly and do not penetrate organic matter well. When using alcohols, it is best to clean an object,

then submerge it in alcohol for the appropriate time. Alcohols are often used in concert with other

disinfectants such as formaldehyde (but see toxicity warning below) or chlorine (2000 ppm chlorine in

alcohol). Alcohol is NOT a registered tuberculocidal or HIV listed disinfectant.

Chlorine compounds

The most commonly used and generally effective disinfectant is sodium hypochlorite (common

household bleach). However, it is a strong oxidizing agent and therefore can be corrosive to metal. A

1:50 dilution, supplying 1000 ppm available chlorine, of the common household product (e.g.,

Chlorox®) is very effective as a general laboratory disinfectant and a 1:10 dilution supplying 5000

ppm available chlorine is effective against spills involving blood or other organic material. Please note

that the presence of high concentrations of protein can inactivate the action of chlorine products.

Dilute hypochlorite solution must be prepared daily to be maximally effective. There are more

concentrated sodium hypochlorite solutions available for industrial use, so please read the product

information carefully to determine the proper dilution.

Table 4: Dilutions of Household Bleach

https://www.fda.gov/medicaldevices/deviceregulationandguidance/reprocessingofreusablemedicaldevices/ucm437347.htm

https://www.fda.gov/medicaldevices/deviceregulationandguidance/reprocessingofreusablemedicaldevices/ucm437347.htm

https://www.epa.gov/pesticide-registration/selected-epa-registered-disinfectants

https://www.epa.gov/pesticide-registration/selected-epa-registered-disinfectants

54

Volume Volume Dilution % Sodium Available

Of Bleach of Water Ratio Hypochlorite Chlorine

(mg/L)

Undiluted 0 1:1 5.25 50,000

1 9 1:10 0.5 5,000

1 99 1:100 0.05 500

Formaldehyde

Formaldehyde is a gas that is available either dissolved in water and methanol as a 37% formaldehyde

solution, known as formalin, or as a solid, paraformaldehyde, that may be melted to release the gas.

Formaldehyde gas is very active against a variety of microorganisms and is used for space

decontamination and to decontaminate biological safety cabinets. Formaldehyde dissolved in water is

active at 1-8% solutions and can be used to decontaminate hard surfaces. However, because

formaldehyde is very irritating at low concentrations (0.1 to 5 ppm) and a known carcinogen, its use

as a hard surface disinfectant is limited to situations in which it is particularly needed. Due to its toxic

effects, there are no EPA-registered disinfectants that contain formaldehyde.

Glutaraldehyde

Glutaraldehyde is usually supplied as a 20% solution and requires activation by the addition of an

alkaline agent prior to use. The activated product may be kept for about two weeks and should be

discarded when turbid. Glutaraldehyde is active against almost all microorganisms, but is toxic,

irritating, and mutagenic and should be used only when necessary. Please follow the manufacturer’s

guidance when using glutaraldehyde-based products, as there are many different formulations that

have been designed for specific uses.

Hydrogen peroxide

Hydrogen peroxide is usually available as a 30% solution. It may be diluted 1:5 for use as a

disinfectant. It is active against a wide array of microorganisms. However, it is an oxidizing agent and

should not be used on aluminum, copper, zinc, or brass. Hydrogen peroxide is unstable at high

temperatures and in light.

Iodine and Iodophors

Iodine and iodophors, compounds in which iodine is combined with a solubilizing or carrier agent, are

general, all-purpose disinfectants with an action similar to that of chlorine products. The appropriate

concentration for iodine-containing products is 75 ppm available iodine for disinfecting work surfaces.

Concentrations may be much higher for other purposes. Like chlorine compounds, the effectiveness of

iodine compounds may be diminished in the presence of protein/organic material. Iodophor

compounds that are used for antisepsis (germicide applied to tissue or skin) are not appropriate for

use as hard surface disinfectants and vice versa. Please read the product material for appropriate

dilutions and applications.

Phenol and phenolic compounds

Phenolic compounds are active at 0.2 to 3% concentrations against all forms of vegetative

microorganisms but not against spores. They have only limited effectiveness against nonlipid viruses.

There are many common phenol-based disinfectants and they should be used according to the

55

manufacturer’s recommendations. Phenol is a hazardous chemical and requires hazard assessment as

well as proper PPE selection and use.

Quatenary ammonium compounds

Compounds in this class are active at concentrations of 0.1 - 2%. They are active against vegetative

bacteria, lipid viruses, but not against bacterial spores, non-lipid viruses, or tubercle bacilli. These

compounds should be used only when a low-level disinfectant is required.

2.7.2 Gasses

Ethylene Oxide (Not currently allowed at NAU)

Inactivates micro-organisms, cellular disruption

Temp. 50-6 °C, Humidity 30-40%

Conc. 400-500 mg/liter

Time 2 hrs at 60°C or 24 hrs at 20°C

Aeration required after cycle for 8 hrs

Biological indicators needed to confirm kill

Suspected carcinogen with explosive properties

Vapor Phase Hydrogen Peroxide

Temp. 4-60°C,> temp. results in > activity

Conc. 30%, less than 10 mg/liter

Non-toxic end products of water and oxygen

Limited to surfaces, no penetration

Corrosive to some materials

Degrades natural rubber and nylon

Chlorine Dioxide gas

Dilute chlorine gas and sodium chlorite, less than 25 mg/liter

Temp. 25-30 °C, pre-humidification required

Limited to surfaces, no penetration

Corrosive to some materials

Mucous membrane irritant

Ozone

Oxidizing properties, inhibits bacterial growth, reacts with amino acids, RNA/DNA

Temp. 25°C

Conc. 2-5 mg/liter

Systems convert oxygen to ozone

Limited penetration

Formaldehyde Gas (from heating paraformaldehyde)

Temp. 20-22°C, humidity 60-85%

56

Conc. 0.3 gm/cu ft. of volume

Time 6-8 hours

Toxic irritant and suspected carcinogen

Limited penetration, primarily surface action

Requires aeration and time for formaldehyde to off-gas, usually 8 hours

2.7.3 Irradiation

Ultraviolet, UV radiation

Virucidal action correlates with shorter wavelengths of the UV spectrum,

250-260 nm.

Mechanism of UV radiation injury attributed to absorption by and resultant damage to nucleic

acids

Due to low energy, the power of penetration is poor.

Dust and thin layers of proteins on surfaces reduce the virucidal activity

Do not rely on biological safety cabinet UV lights to disinfect surfaces.

Ionizing radiation

Penetrates products and micro-organisms

Releases high energy, disrupts cells, DNA & RNA

Damage done to cell membrane and other cellular structures

Cobalt 60 and Cesium 137

Electron Beam

Limited penetration

Accelerator generates high energy electrons

Microwaves

Thermal and non-thermal activity

Still under study for the most part

2.7.4 Disinfectants Bibliography

The following materials were consulted or used in this chapter: Disinfection, Sterilization, and Preservation. Fourth edition. 1991. Seymour S. Block ed., Lea & Febiger, Philadelphia

Laboratory Biosafety Manual. Second edition. 1993. World Health Organization, Geneva.

Laboratory Safety: Principles and practices. Second edition. 1995. Diane O. Fleming et al. eds. ASM Press, Washington

DC

Manual of Clinical Microbiology. Fifth edition. 1991. Albert Balows ed., ASM, Washington DC

Prudent Practices in the Laboratory: Handling and disposal of chemicals. 1995 National Research Council. National

Academy Press, Washington

57

Table 5: Summary and Comparison of Liquid Disinfectants

Commonly used disinfectants, recommended when appropriate.

Class Recommended Use How They Work Advantages Disadvantages Comments/Hazards Examples

70% Isopropyl

alcohol

solution

• Cleaning some

instruments

• Cleaning skin

• Changes protein

structure of

microorganism

• Presence of water

assists with killing action

• Fairly inexpensive • <50% solution not very

effective

• Not active when organic

matter present

• Not active against certain

types of viruses

• Evaporates quickly - contact

time not sufficient for killing

• Flammable

• Eye Irritant

• Toxic

Chlorine

compounds • Spills of human body

fluids

• Bactericidal - Good

• Fungicidal - Good

• Sporicidal - Good at

>1000 ppm Sodium

Hypochlorite

• Free available chlorine

combines with contents

within microorganism -

reaction byproducts cause

its death

• Need 500 to 5000 ppm

• Produce chemical

combination with cell

substances

• Depend upon release of

hypochlorous acid

• Kill hardy viruses

(e.g., hepatitis)

• Kill a wide range of

organisms

• Inexpensive

• Penetrates well

• Relatively quick

microbial kill

• May be used on food

prep surfaces

• Corrode metals such as

stainless, aluminum

• Organics may reduce activity

• Increase in alkalinity

decreases bactericidal property

• Unpleasant taste and odor

• Tuberculocidal - with

extended contact time

• Follow spill procedure

and dilution instructions

• Make fresh solutions

before use

• Eye, skin, and

respiratory irritant

• Corrosive

• Toxic

• Bleach

solutions

(sodium

hypochlorite)

• Clorox

• Cryosan

• Purex

Gluteraldehyde • Bactericidal - Good


• Tuberculocidal -

Excellent

• Virucidal - Good

• Sporicidal - Good

• Coagulates cellular

proteins

• Non-staining,

relatively non-corrosive

• Useable as a sterilant

on plastics, rubber,

lenses, stainless steel,

and other items that

can’t be autoclaved

• Not stable in solution

• Has to be in alkaline solution

• Inactivated by organic

material

• Eye, skin and

respiratory irritant.

• Sensitizer

• Toxic

• Calgocide

14

• Cidex

• Vespore

58

Table 5: Summary and Comparison of Liquid Disinfectants

Class Recommended

Use

How They Work Advantages Disadvantages Comments/Hazards Examples

Iodophors

(iodine with

carrier)

• Disinfecting some

semi-critical medical

equipment

• Bactericidal - Very

Good

• Fungicidal - Excellent

• Virucidal - Excellent

• Free iodine enters

microorganism and binds

with its cellular

components

• Carrier helps penetrate

soil/fat

• Need 30 to 50 ppm

• Probably by disorder of

protein synthesis due to

hindrance and/or

blocking of hydrogen

bonding

• Kill broad range of

organisms

• Highly reactive

• Low tissue toxicity

• Kill immediately rather than

by prolonged period of stasis

• Not affected by hard water

• May be used on food prep

surfaces

• May stain plastics or

corrode metal

• May stain skin/laundry

• Stains most materials

• Odor

• Some organic and

inorganic substances

neutralize effect

• Tuberculocidal - with

extended contact time

• Sporicidal - some

• Dilution critical - follow

directions!

• Use only EPA-registered

hard surface iodophor

disinfectants

• Don’t confuse skin

antiseptic iodophors for

disinfectants

• Skin and eye irritant

• Corrosive

• Toxic

• Bactergent

• Hy-Sine

• Ioprep

• Providone-

iodine;

betadine

•

Wescodyne

Phenolic

Compounds • Bactericidal -

Excellent

• Fungicidal - Excellent

• Tuberculocidal -

Excellent

• Virucidal - Excellent

• Gross protoplasmic

poison

• Disrupts cell walls

• Precipitates cell

proteins

• Low concentrations

inactivate essential

enzyme systems

• Nonspecific concerning

bactericidal and fungicidal

action

• When boiling water would

cause rusting, the presence of

phenolic substances produces

an anti-rusting effect

• Unpleasant odor

• Some areas have

disposal restrictions

• Effectiveness reduced

by alkaline pH, natural

soap, or organic material

• Sporicidal - NO


• Sensitizer

• Corrosive

• Toxic

• Hil-Phene

• Lph

• Metar

• Vesphene

Quaternary

ammonium

compounds

(QUATS)

• Ordinary

housekeeping

(e.g., floors,

furniture, walls)

• Bactericidal -

Excellent


• Virucidal - Good (not

as effective as phenols)

• Affect proteins and cell

membrane of

microorganism

• Release nitrogen and

phosphorous from cells

• Contain a detergent to help

loosen soil

• Rapid action

• Colorless, odorless

• Non-toxic, less corrosive

• Highly stable

• May be used on food prep

surfaces

• Do not eliminate

spores, TB bacteria,

some viruses

• Effectiveness

influenced by hard water

• Layer of soap

interferes with action

• Select from EPA list of

hospital disinfectants


• Toxic

• Coverage

258

• End-Bac

• Hi Tor

Barbara Fox Nellis (8-23-96)

59

Table 6: Summary of Practical Disinfectants

Quatenary Ammonium

Compounds

Phenolic

Compounds

Chlorine

Compounds Iodophor Ethyl Alcohol

Isopropyl

Alcohol Formaldehyde Glutaraldehyde

Inactivates

Vegetative Bacteria + + + + + + + +

Lipoviruses + +a + + +a + + +

Nonlipid Viruses - - + + - - + +

Bacterial Spores - - + + - - + +

Treatment Requirements

Use Dilution 0.1-2.0% 1.0-5.0% 500ppmb 25-1600ppmb 70-85% 70-85% 0.2-0.8% 2%

Contact Time-minutes

Lipovirus 10 10 10 10 10 10 10 10

Broad Spectrum NE NE 30 30 NE NE 30 30

Important Characteristics

Effective Shelf Life

> 1 weekc

+ + - + + + + +

Corrosive - + + + - - - -

Flammable - - - - + + - -

Explosion Potential - - - - - - - -

Inactivated by organic

matter

+ - + + - - - -

Skin Irritant + + + + - - + +

Eye Irritant + + + + + + + +

Respiratory Irritant - - + - - - + +

Toxicd + + + + + + + +

Applicability

Waste Liquids - - + - - - - -

Dirty Glassware + + + + + + + +

Equipment, Surface

Decontamination

+ + + + + + + +

Proprietary Productse A-33, CDQ, End-Bac,

Hl-Tor, Mikro-Quat

Hil-Phene,

Metar, Mikro-

Bac, O-Syl

Chloramine

T, Clorox,

Purex

Hy-Sine, Ioprep,

Mikroklene,

Wescodyne

Sterac Cidex

Source: Adapted from Laboratory Safety Monograph, A Supplement to the NIH Guidelines for Recombinant DNA Research. National Institutes of Health, Office of

Research Safety, National Cancer Institute, and the Special Committee of Safety and Health Experts, Bethesda, Maryland. January 1989: 104-105

+ Yes a Variable results depending on virus d By skin or mouth or both

- No b Available Halogen e Space limitations preclude listing all products available. Individual listings (or omissions)

NE Not Effective c Protected from light and air do not imply endorsement (or rejection) of any product by the National Institutes of Health.

60

Table 7: Reprocessing Methods for Equipment Used in the Health Care Setting

Sterilization: Destroys: All forms of microbial life including high numbers of bacterial spores.

Methods:

Steam under pressure (autoclave), gas (ethylene oxide), dry heat or immersion in an approved chemical “sterilant” (e.g., US Environmental Protection

Agency-approved) for prolonged period of time, e.g., 6-10 hours or according to manufacturer’s instructions. Note: Liquid chemical “sterilants” should

be used only on those instruments that are impossible to sterilize or disinfect with heat.

Use:

For those instruments or devices that penetrate skin or contact normally sterile areas of the body, e.g., scalpels, needles, etc. Disposable invasive

equipment eliminates the need to reprocess these types of items. When indicated, however, arrangements should be made with a health-care facility for

reprocessing of reusable invasive instruments.

High-Level

Disinfection:

Destroys: All forms of microbial life except high numbers of bacterial spores.

Methods: Hot water pasteurization (80-100°C, 30 minutes) or exposure to an approved (e.g., US EPA-approved) “sterilant” chemical as above, except for a short

exposure time (10-45 minutes or as directed by the manufacturer).

Use: For reusable instruments or devices that come into contact with mucous membranes (e.g., laryngoscope blades, endotracheal tubes, etc.).

Intermediate-

Level

Disinfection:

Destroys: Mycobacterium tuberculosis, vegetative bacteria, most viruses and most fungi, but does not kill bacterial spores.

Methods:

Approved (e.g., US EPA-approved) “hospital disinfectant” chemical germicides that have a label claim for tuberculocidal activity; commercially

available hard-surface germicides or solutions containing at least 500 ppm free available chlorine (a 1:10 dilution of common household bleach- about ¼

cup bleach per gallon of tap water).

Use: For those surfaces that come into contact only with intact skin, e.g., stethoscopes, blood pressure cuffs, splints, etc., and have been visibly contaminated

with blood or bloody body fluids. Surfaces must be pre-cleaned of visible material before the germicidal chemical is applied for disinfection.

Low-Level

Disinfection:

Destroys: Most bacteria, some viruses, some fungi, but no Mycobacterium tuberculosis or bacterial spores.

Methods: Approved (e.g., US EPA-approved) “hospital disinfectants” (no label claim for tuberculocidal activity).

Use: Use these excellent cleaning agents for routine housekeeping or removal of soiling in the absence of visible blood contamination.

Environmental

Disinfection: Environmental surfaces that have become soiled should be cleaned and disinfected using any cleaner or disinfectant that is intended for environmental

use. Such surfaces include floors, woodwork, ambulance seats, countertops, etc.

Important: To assure effectiveness of any sterilization or disinfection process, equipment and instruments must first be thoroughly cleaned of all visible soil.

61

2.8 Shipment of Biological Materials

The following regulations apply to the packaging and shipment of biological materials:

U.S. Department of Transportation, 49 CFR Parts 171-180 and amendments

U.S. Public Health Service, 42 CFR Part 72, Interstate Shipment of Etiologic Agents

U.S. Department of Labor, Occupational Safety and Health Administration, 29 CFR Part 1910.1030,

Bloodborne Pathogens

International Air Transport Association (IATA), Dangerous Goods Regulations

U.S. Postal Service, 39 CFR Part 111, Mailability of Etiologic Agents, Mailability of Sharps and Other

Medical Devices, and Publication 52, Acceptance of Hazardous, Restricted or Perishable Matter

International Civil Aviation Organization, Technical Instructions for the Safe Transport of

Dangerous Goods by Air

United Nations, Recommendations of the Committee of Experts on the Transportation of Dangerous

Goods

All North American airlines and FedEx, the largest shipper of infectious materials, use the IATA

regulation (also referred to as the Dangerous Goods Regulation or DGR) as their standard. Meeting the

conditions of this standard will ensure meeting the provisions of the other US regulations.

General information is provided below, but please contact the EH&S Office of Biological Safety for

specific information. Note that for any biological materials for which a state or federal permit or license is

required, registration with the EH&S Office of Biological Safety is also required.

Any NAU faculty/staff, regardless of job or task, must be trained to package, ship or receive packages of

materials/samples classified by regulation as “dangerous goods”. This training is available from EH&S.

Regulation requires retraining every three years OR whenever the regulations change.

2.9 Laboratory Spills

A spill of biological materials that takes place in the open laboratory may result in the exposure of

laboratory personnel to infectious materials and is a serious problem. Every effort should be taken to

prevent spilling materials.

A spill poses less of a problem if it happens inside a biological safety cabinet, provided splattering to the

outside of the cabinet does not occur. Direct application of concentrated liquid disinfectant and a

thorough wipe down of the internal surfaces of the cabinet will usually be effective for decontaminating

the work zone, but gaseous sterilants will be required to disinfect the interior sections of the cabinet and

HEPA filter(s) if they become contaminated.

2.9.1 Spill in the Open Laboratory

Advance preparation for management of a spill is essential. A “spill kit” may include the following:

• Leak-proof containers

• Forceps

• Paper towels

• Disinfectant

• Respirators, if necessary

• Rubber gloves and other personal protective equipment

• Absorbent powder or other material

62

• Red biohazard bag

If potentially hazardous biological material is spilled in the laboratory, avoid inhaling any aerosolized

material by holding your breath and leaving the laboratory. Warn others in the area by posting signs and

go directly to wash or change room area. If clothing is known (or suspected) to be contaminated, remove

the clothing with care, folding the contaminated area inward. Discard the clothing into a bag or place the

clothing directly in an autoclave. Wash all potentially contaminated body areas as well as the arms, face

and hands. Shower if facilities are available. Re-entry into the laboratory should be delayed for a period

of at least thirty minutes to allow reduction of the aerosol generated by the spill.

Protective clothing should be worn when entering the laboratory to clean the spill area. Wear rubber

gloves, autoclavable or disposable footwear, an outer garment, and a respirator equipped with a HEPA

filter, as deemed appropriate by the size and nature of the spill. If the spill was on the floor, do not use a

surgical gown that may trail on the floor when bending down. Take the “spill kit” into the laboratory

room, place a waste container near the spill, and transfer large fragments of material into it. Cover the

spill with paper towels or another appropriate disinfectant. Using a freshly prepared hypochlorite

containing 5000 ppm (10% by volume) available chlorine, iodophor solution containing 1600 ppm

iodine, or other appropriate EPA registered disinfectant, carefully pour the disinfectant around and onto

the visible spill. Avoid splashing and work from the outside toward the center. Allow 30 minutes' contact

time. Use paper or cloth towels to wipe up the disinfectant and spill, working toward the center of the

spill. Discard all towels and other clean up materials into a waste container as they are used. Wipe the

outside of the discard containers, especially the bottom, with a towel soaked in a disinfectant. Place the

spill waste and other materials in an autoclave and sterilize. Alternately, place all materials in the

appropriate biomedical waste system for incineration. Remove shoes or shoe covers, outer clothing,

respirator, and gloves and sterilize by autoclaving. Wash hands, arms and face, or if possible, shower.

If gaseous decontamination of the room is required, contact the Office of Biological Safety.

2.9.2 Spill in a Biological Safety Cabinet

A spill that is confined to the interior of the biological safety cabinet should present little or no hazard to

personnel in the area. However, chemical disinfection procedures should be initiated at once while the

cabinet ventilation system continues to operate to prevent escape of contaminants from the cabinet.

While wearing gloves and any other appropriate PPE, cover the spill with paper towels or other

absorbent material. Pour appropriate disinfectant over the spill, starting from the outside of the spill and

working inward. Wait the appropriate contact time. Use paper or cloth towels to wipe up the disinfectant

and spill, working toward the center of the spill. Discard all towels and other clean up materials into a

waste container as they are used. Spray or wipe any potentially contaminated walls, work surfaces, and

equipment with a disinfectant. A disinfectant with a detergent has the advantage of detergent activity that

will help clean the surfaces by removing both dirt and microorganisms. A suitable disinfectant is a 3%

solution of an iodophor such as Wescondyne or a 1:100 dilution of household bleach (e.g., Chlorox) with

0.7% nonionic detergent. If the spill penetrated the front grille, use sufficient disinfectant solution to

ensure that the drain pans and catch basins below the work surface contain the disinfectant. Lift the front

exhaust grill and tray and wipe all surfaces. Wipe the catch basin and drain the disinfectant into a

container. The disinfectant, gloves, wiping cloth and sponges should be discarded into an autoclave pan

and autoclaved.

The above procedure will not disinfect the filters, blower, air ducts, or other interior parts of the cabinet.

If the entire interior of the cabinet needs to be sterilized, contact the EH&S Office of Biological Safety

for the name of the current contractor.

63

2.10 Biological Safety Cabinets

The following is excerpted from Primary Containment for Biohazards: Selection, Installation and Use of Biological Safety

Cabinets, 1995, Centers for Disease Control & Prevention/National Institutes of Health

This chapter presents information on the selection,

function, and use of biological safety cabinets (BSCs),

which are the primary means of containment

developed for working safely with infectious

microorganisms. BSCs are designed to provide

personnel, environment, and product protection when

appropriate practices and procedures are followed.

Three kinds of biological safety cabinets, designated as

Class I, II, and III, have been developed to meet

varying research and clinical needs.

Biological safety cabinets use high efficiency

particulate air (HEPA) filters in their exhaust and/or

supply systems.

2.10.1 Biological Safety Cabinets (BSCs)

The similarities and differences in protection offered by the various classes of biological safety cabinets

are reflected in Table 1.

The Class I BSC

This type of cabinet is not for aseptic or sterile technique. The Class I BSC provides personnel and

environmental protection, but no product protection. It is similar in air movement to a chemical fume

hood, but has a HEPA filter in the exhaust system to protect the environment.

The Class II BSC

The Class II (Types A and B) biological safety cabinets provide personnel, environmental, and product

protection. Air flow is drawn around the operator into the front grille of the cabinet, which provides

personnel protection. In addition, the downward laminar flow of HEPA-filtered air provides product

protection by minimizing the chance of cross-contamination along the work surface of the cabinet.

Because cabinet air has passed through the exhaust HEPA filter, it is contaminant-free (environmental

protection), and may be recirculated back into the laboratory (Type A BSC) or ducted out of the building

(Type B BSC).

The Class II, Type A BSC

An unducted Class II Type A BSC is not to be used for work involving volatile or toxic chemicals. The

buildup of chemical vapors in the cabinet (by recirculated air) and in the laboratory (from exhaust air)

could create health and safety hazards. Generally, BSCs are not for use with chemicals. Small quantities

of chemicals and chemotherapeutic agents may be used in ducted, 100% BSCs.

It is possible to duct the exhaust from a Type A cabinet out of the building. However, it must be done in

a manner that does not alter the balance of the cabinet exhaust system, thereby disturbing the internal

cabinet air flow. The typical method of ducting a Type A cabinet is to use a “thimble”, or canopy unit,

which maintains a small opening (usually 1 inch) around the cabinet exhaust filter housing. The

64

volume of the exhaust must be sufficient to maintain the flow of room air into the space between the

thimble unit and the filter housing (contact manufacturers for any additional specifications). The thimble

must be removable or be designed to allow for operational testing of the cabinet. The performance of a

cabinet with this exhaust configuration is unaffected by fluctuations in the building exhaust system.

“Hard-ducting” (i.e., direct connection) of Class II Type A cabinets to the building exhaust system is not

recommended. The building exhaust system must be precisely matched to the airflow from the cabinet in

both volume and static pressure. However, fluctuations in air volume and pressure that are common to all

building exhaust systems make it difficult, if not impossible, to match the airflow requirements of the

cabinet.

The Class II, Type B1 BSC

Some biomedical research requires the use of small quantities of certain hazardous chemicals, such as

carcinogens. The powdered form of these carcinogens should be weighed or manipulated in a chemical

fume hood or a static-air glove box. Carcinogens used in cell culture or microbial systems require both

biological and chemical containment.

Type B1 cabinets must be hard-ducted to their own dedicated exhaust system. As indicated earlier,

blowers on laboratory exhaust systems should be located at the terminal end of the ductwork. A failure in

the building exhaust system may not be apparent to the user, as the supply blowers in the cabinet will

continue to operate. A pressure-independent monitor should be installed to sound an alarm and shut off

the BSC supply fan, should failure in exhaust airflow occur. Since all cabinet manufacturers do not

supply this feature, it is prudent to install a sensor in the exhaust system as necessary. To maintain critical

operations, laboratories using Type B BSCs should connect the exhaust blower to the emergency power

supply.


This BSC is a total-exhaust cabinet; no air is recirculated within it. This cabinet provides simultaneous

primary biological and chemical containment.

Should the building or cabinet exhaust fail, the cabinet will be pressurized, resulting in a flow of air from

the work area back into the laboratory. Cabinets built since the early 1980's usually have an interlock

system installed by the manufacturer to prevent the supply blower from operating whenever the exhaust

flow is insufficient. Presence of such an interlock system should be verified; systems can be retrofitted if

necessary. A pressure-independent device should monitor exhaust air movement.


This biological safety cabinet is a ducted Type A cabinet. All positive pressure contaminated plenums

within the cabinet are surrounded by a negative air pressure plenum. Thus, leakage in a contaminated

plenum will be into the cabinet and not into the environment.

The Class III BSC

The Class III biological safety cabinet was designed for work with biosafety level 4 microbiological

agents, and provides maximum protection to the environment and the worker. It is a gas-tight enclosure

with a non-opening view window.

Long, heavy-duty rubber gloves are attached in a gas-tight manner to ports in the cabinet and allow for

manipulation of the materials isolated inside. Although these gloves restrict movement, they prevent the

65

user's direct contact with the hazardous materials. The trade-off is clearly on the side of maximizing

personal safety. Depending on the design of the cabinet, the supply HEPA filter provides particulate-free,

albeit somewhat turbulent, airflow within the work environment.

2.10.2 Horizontal Laminar Flow “Clean Bench”

Horizontal laminar flow clean air benches are not BSCs. They discharge HEPA-filtered air across the

work surface and toward the user. These devices only provide product protection. They can be used for

certain clean activities, such as the dust-free assembly of sterile equipment or electronic devices. These

benches should never be used when handling cell culture materials or drug formulations, or when

manipulating potentially infectious materials. The worker can be exposed to materials (including

proteinaceous antigens) being manipulated on the clean bench, which may cause hypersensitivity.

Horizontal clean air benches should never be used as a substitute for a biological safety cabinet in

research, biomedical or veterinary laboratories and/or applications.

2.10.3 Vertical Laminar Flow “Clean Bench”

Vertical laminar flow clean benches also are not BSCs. They may be useful, for example, in hospital

pharmacies when a clean area is needed for preparation of intravenous drugs. While these units generally

have a sash, the air is usually discharged into the room under the sash, resulting in the same potential

problems as the horizontal laminar flow clean benches.

2.10.4 Operations within a Class II BSC

Laboratory Hazards

Many common procedures conducted in BSCs may create splatter or aerosols. Good microbiological

techniques should always be used when working in a biological safety cabinet. For example, techniques

to reduce splatter and aerosol generation will minimize the potential for exposure to personnel from

infectious materials manipulated within the cabinet. Class II cabinets are designed so that horizontally

nebulized spores will be captured by the downward flowing cabinet air within fourteen inches of travel.

Therefore, as a general rule of thumb, keeping clean materials at least one foot away from aerosol-

generating activities will minimize the potential for cross-contamination.

The general workflow should be from “clean to contaminated (dirty)”. Materials and supplies should be

placed in such a way as to limit the movement of “dirty” items over “clean” ones.

Several measures can be taken to reduce the chance for cross-contamination when working in a BSC.

Work at least 10” back from the front edge and never cover the front grille. Opened tubes or bottles

should not be held in a vertical position. Investigators working with Petri dishes and tissue culture plates

should hold the lid above the open sterile surface to minimize direct impact of downward air. Bottle or

tube caps should not be placed on the toweling. Items should be recapped or covered as soon as possible.

Open flames are not permitted in the near microbe-free environment of a BSC. On an open bench,

flaming the neck of a culture vessel will create an upward air current that prevents microorganisms from

falling into the tube or flask. An open flame in a BSC, however, creates turbulence that disrupts the

pattern of air supplied to the work surface. When deemed absolutely necessary, touch-plate microburners

equipped with a pilot light to provide a flame on demand may be used. Internal cabinet air disturbance

and heat buildup will be minimized. The burner must be turned off when work is completed. Small

electric “furnaces” are available for decontaminating bacteriological loops and needles and are preferable

66

to an open flame inside the BSC. Disposable sterile loops should be used to eliminate the need for heat or

flame.

Aspirator bottles or suction flasks should be connected to an overflow collection flask containing

appropriate disinfectant, and to an in-line HEPA or equivalent filter. This combination will provide

protection to the central building vacuum system or vacuum pump, as well as to the personnel who

service this equipment. Inactivation of aspirated materials can be accomplished by placing sufficient

chemical decontamination solution into the flask to kill the microorganisms as they are collected. Once

inactivation occurs, liquid materials can be disposed of appropriately as noninfectious waste.

Investigators must determine the appropriate method of decontaminating materials that will be removed

from the BSC at the conclusion of the work. When chemical means are appropriate, suitable liquid

disinfectant should be placed into the discard pan before work begins. Items should be introduced into

the pan with minimum splatter, and allowed appropriate contact time as per manufacturer's instructions.

Alternatively, liquids can be autoclaved prior to disposal. Contaminated items should be placed into a

biohazard bag or discard tray inside the BSC. Water should be added to the bag or tray prior to

autoclaving.

When a steam autoclave is to be used, contaminated materials should be placed into a biohazard bag or

discard pan containing enough water to ensure steam generation during the autoclave cycle. The bag

should be taped shut or the discard pan should be covered in the BSC prior to removal to the autoclave.

The bag should be transported and autoclaved in a leak-proof tray or pan.

2.10.5 Decontamination

Surface Decontamination

All containers and equipment should be surface decontaminated and removed from the cabinet when

work is completed. At the end of the workday, the final surface decontamination of the cabinet should

include a wipe-down of the work surface, the cabinet's sides and back, and the interior of the glass. If

necessary, the cabinet should also be monitored for radioactivity and decontaminated when necessary.

Investigators should remove their gloves and gowns and wash their hands as the final step in safe

microbiological practices.

Small spills within the BSC can be handled immediately by removing the contaminated absorbent paper

toweling and placing it into the biohazard bag. Any splatter onto items within the cabinet, as well as the

cabinet interior, should be immediately wiped with a towel dampened with decontaminating solution.

Gloves should be changed after the work surface is decontaminated and before placing clean absorbent

toweling in the cabinet. Hands should be washed whenever gloves are changed or removed.

Spills large enough to result in liquids flowing through the front or rear grilles require more extensive

decontamination. All items within the cabinet should be surface decontaminated and removed. After

ensuring that the drain valve is closed, decontaminating solution can be poured onto the work surface and

through the grille(s) into the drain pan.

Thirty minutes is generally considered an appropriate contact time for decontamination, but this varies

with the disinfectant and the microbiological agent. Manufacturer's directions should be followed. The

spilled fluid and disinfectant solution on the work surface should be absorbed with paper towels and

discarded into a biohazard bag. The drain pan should be emptied into a collection vessel containing

disinfectant. A flexible tube should be attached to the drain valve and be of sufficient length to allow the

67

open end to be submerged in the disinfectant within the collection vessel. This procedure serves to

minimize aerosol generation. The drain pan should be flushed with water and the drain tube removed.

Should the spilled liquid contain radioactive material, a similar procedure can be followed. The EH&S

Radiation Safety Officer should be contacted for specific instructions. See contact information in the front

of this manual.

Gas Decontamination

BSCs that have been used for work involving infectious materials must be decontaminated before HEPA

filters are changed, internal repair work is done or before a BSC is relocated. The most common

decontamination method uses formaldehyde gas, although more recently, hydrogen peroxide vapor has

been used successfully. This environmentally benign vapor is useful in decontaminating HEPA filters,

isolation chambers, and centrifuge enclosures. Call the EH&S Office of Biological Safety for the current

BSC vendor who does the gas decontaminations and certifications. All BSCs must be re-certified

following any gas decontamination, maintenance or relocation.

2.10.6 Engineering Requirements

Ultraviolet Lamps

Ultraviolet (UV) lamps are not required in BSCs. If installed, UV lamps must be cleaned weekly to

remove any dust and dirt that may block the germicidal effectiveness of the ultraviolet light. The lamps

should be checked periodically with a meter to ensure that the appropriate intensity of UV light is being

emitted. UV lamps must be turned off when the room is occupied to protect eyes and skin from UV

exposure, which can burn the cornea and cause skin cancer. Do not depend on UV lamps to disinfect the

area.

BSC Placement

The ideal location for the biological safety cabinet is remote from the entry (e.g., the rear of the laboratory

away from traffic), since people walking parallel to the face of a BSC can disrupt the air curtain. The air

curtain created at the front of the cabinet is quite fragile, amounting to a nominal inward and downward

velocity of 1 mph. Open windows, air supply registers, or laboratory equipment that creates air

movement (e.g., centrifuges, and vacuum pumps) should not be located near the BSC. Similarly,

chemical fume hoods must not be located close to BSCs.

HEPA Filters

HEPA filters, whether part of a building exhaust system or part of a cabinet, will require replacement

when they become so loaded that sufficient air flow can no longer be maintained. Filters must be

decontaminated before removal.

Certification of BSCs

All BSCs must be certified (according to a National Sanitation Foundation standard) annually according

to NAU policy. Please contact the EH&S Office of Biological Safety for the name and phone number of

the current contractor performing this service. Prices and quality vary widely, so only BSO approved

contractors may be used.

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3.0 PROGRAMS

3.1 Occupational Health and Safety (Occupational Health Program)

Occupational health and safety is of utmost importance to the NAU IACUC. According to the Public

Health Service, an occupational health program is required for institutions that employ personnel who

have animal contact. The NAU Occupational Health Program is overseen by the NAU Institutional

Animal Care and Use Committee (IACUC). Regulations require that the IACUC monitor personnel

occupational health programs and training related to animal use at NAU. Occupational health programs

must include training in basic safety techniques and information about hazards which will be encountered

in each particular job. Hazardous materials must be stored safely and in compliance with MSDS and

OSHA regulations. Posted signs must warn of hazards present. The NAU program is designed to protect

all personnel from occupational exposure to conditions that may result in animal related illnesses.

Responsibilities Under the Program

All individuals who engage in animal-related work will participate in the university occupational health

program. As the initial step in this program, individuals will complete the "Occupational Health

Questionnaire" found at the IACUC website http://www.nau.edu/research/compliance/animal-

care/Certification-Training/. The University Veterinarian will review the questionnaire, make any

comments which may highlight animal work related concerns, and forward the form to health care

providers at Campus Health Services. At the discretion of Campus Health personnel, the individual will

be scheduled for a brief interview or examination, where potential health-risks will be discussed relative

to the individual's health status and job requirements. All individuals who work with animals will be

current on their tetanus vaccine. Serology and/or vaccination for rabies, Hepatitis B, toxoplasmosis,

measles, and skin testing for tuberculosis may be required for work with certain species or protocols.

Vaccination for rabies will be required to work with wild or random-source bats, skunks, and carnivores.

Tuberculosis testing will be required for work with all non-human primates. This program is provided at

no cost to the employee.

Principle Investigators are responsible for ensuring that all personnel (including employees, students,

colleagues, collaborators, and volunteers) involved with their IACUC-approved project are given this

program information and that they receive the appropriate immunizations and tests, etc. Investigators

who do not respond to requests for registration may have their animal use approval rescinded by the

IACUC.

Supervisors are responsible for providing information to animal care technicians, veterinarians, and

associated animal workers (e.g., workers in Animal Care Services) who work with animals but are not on

a specific research project.

The University Veterinarian and/or health care providers at Campus Health Services will conduct an

educational component of the occupational health program as appropriate. Topics to be discussed include

zoonoses relative to the protocol, allergy risk, possible risks to immunocompromised individuals, and use

of protective equipment and supplies such as gloves, eye protection, masks, lab coats, scrubs, respirators,

head covers, boots, and heat resistant gloves chemical resistant gloves.

An animal exposure case involves mucous membrane contact with animal fluids or excreta, ingestion of

animal fluids or excreta, bites, scratches, etc. Emergency medical care for minor animal exposure cases

is available at Campus Health Services during business hours. For more serious cases or during non-

business hours, care is available at the Flagstaff Medical Center Emergency Room. In cases where

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individuals may be exposed to non-routine or exotic diseases, it is necessary to have emergency care

sheets to provide basic information for medical personnel. Such information may include MSDS's, sheets

copied from the "Control of Communicable Diseases Manual" (https://www.apha.org/ccdm), or

monographs written by disease experts. Such sheets should be readily available so affected individuals

may easily take them when seeking emergency care. MSDS's for chemicals used in laboratories must

also be readily available.

3.2 Bloodborne Pathogen Program

In December 1991, OSHA published the final rule covering

occupational exposure to bloodborne pathogens. This was adopted

by the state of Arizona and written into the Arizona Administrative

Code in January 1993. NAU instituted its program in the spring of

1993.

The rule requires that those who handle human blood or other

potentially infectious (human) materials as part of their job duties

participate in an employer-generated program. This program shall

include development and annual review of a site specific Exposure Control Plan, annual training

regarding exposures, offer of hepatitis B vaccinations free-of-charge, and post-exposure health care

services.

Northern Arizona University manages this program through the EH&S Office of Biological Safety. The

BBP program materials and information is available on our website at

https://nau.edu/research/compliance/environmental-health-and-safety/training/.

In addition to the program materials, the EH&S Office of Biological Safety provides training sessions

upon request.

Hepatitis B vaccinations are given by the Campus Health Services. Official program medical records are

kept by Campus Health Services, and documentation of vaccination acceptance or declination is

maintained by the EH&S Office of Biological Safety. There is a requirement that Flagstaff Medical

Center staff or other providers send vaccination and post-exposure records to Campus Health Services

for record-keeping purposes.

The EH&S Office of Biological Safety monitors for NAU compliance by requiring training and

vaccination documentation and by confirming BBP participation during the annual laboratory safety

inspection, which is conducted in each laboratory annually.

For further information, please call the EH&S Office of Biological Safety.

4.0 MEDICAL SURVEILLANCE

4.1 Immunoprophylaxis

Northern Arizona University follows the recommendations of the Centers for Disease Control and

Prevention (CDC) and the Public Health Service Advisory Committee for Immunization Practices

(ACIP) for vaccination of at-risk personnel.

Currently, NAU has written policies regarding the need for hepatitis B, anthrax, plague, tetanus, rabies,

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70

and vaccinia vaccination for certain at-risk personnel. Other vaccinations may be required or

recommended, as needed. Particular attention is given to individuals who are or may become

immunocompromised, as recommendations for vaccine administration may be different than for

immunologically competent adults. Various other factors may be taken into account such as pregnancy,

history of allergy, or HIV status.

When considering the need for immunization, a risk assessment will be conducted by the health care

provider in conjunction with information regarding the experimental agent provided by the EH&S Office

of Biological Safety.

4.1.1 Prophylactic Recommendations for Working with Infectious Agents

Please note, the following is derived from the CDC publication, Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, 2007 and the CDC publication, “Update on Adult Immunization: Recommendations of the Immunization Advisory Committee (ACIP),” 1991, MMWR vol. 40.

Bacterial agents

1. Bacillus anthracis

A licensed vaccine is available through the CDC. However, immunization of laboratory personnel is

not recommended unless frequent work with clinical specimens or diagnostic cultures is anticipated

and in settings in which production quantities, high concentrations, or procedures with a high

potential for aerosol (all BSL-3 work) are encountered. In these settings, vaccination is

recommended for all persons working with the agent, all persons working in the laboratory room

where the cultures are handled, and all persons working with infected animals.

2. Clostridium botulinum

A pentavalent (ABCDE) botulism toxoid is available through the CDC as an investigational new

drug (IND). This toxoid is recommended for personnel working with cultures of C. botulinum or its

toxins.

3. Clostridium tetani

The adult diphtheria-tetanus toxoid is highly recommended for laboratory personnel who manipulate

cultures or toxins and is required by the NAU Animal Care Program for all NAU personnel who

work with animals. This immunization should be given every 10 years.

4. Corynebacterium diphtheria

The adult diphtheria-tetanus toxoid every 10 years is recommended for laboratory personnel who

work with this agent.

5. Francisella tularensis

Vaccination for F. tularensis is available and should be considered for personnel working with

infectious material or infected rodents. It is recommended for persons working in or entering

laboratories or animal rooms where cultures or animals are maintained.

6. Neisseria meningitidis

Vaccines for N. meningitidis are available and should be considered for personnel regularly working

with infectious materials. The reader is advised to consult the current recommendations of the

Advisory Committee on Immunization Practices (ACIP) published in the CDC Morbidity and

Mortality Weekly Report (MMWR) for recommendations for vaccination against N. meningitidis.

7. Salmonella typhi

Vaccines for S. typhi are available and should be considered for personnel regularly working with

potentially infectious materials. The reader is advised to consult the current recommendations of the

Advisory Committee on Immunization Practices (ACIP) published in the CDC Morbidity and

Mortality Weekly Report (MMWR) for recommendations for vaccination against S. typhi.

8. Yersinia pestis

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Vaccination for Y. pestis is available and should be considered for personnel working with

infectious materials or infected rodents. For example: laboratory personnel who routinely perform

procedures that involve viable Y. pestis; and persons (e.g., mammologists, ecologists, and other field

workers) who have regular contact with wild rodents or their fleas in areas in which plague is

enzootic or epizootic. The reader is advised to consult the current recommendations of the Advisory

Committee on Immunization Practices (ACIP) published in the CDC Morbidity and Mortality

Weekly Report (MMWR) for information on vaccination against Y. pestis.

Rickettsial agents

1. Coxiella burnetii

An investigational Phase I, Q fever vaccine (IND) is available on a limited basis from the Special

Immunizations Program (301-619-4653) of the USAMRIID, Fort Detrick, Maryland, for at-risk

personnel under a cooperative agreement with the individual's requesting institution. The use of this

vaccine should be restricted to those who are at high risk of exposure and who have no

demonstrated sensitivity to Q fever antigen.

Viral agents

1. Hepatitis A virus

A vaccine is available and recommended for those who work with hepatitis A virus.

2. Hepatitis B virus

A vaccine is available and highly recommended for those who work with hepatitis B virus. The

hepatitis B vaccine is recommended for those who have occupational exposure to human blood or

other potentially infectious human materials, according to the OSHA rule and the NAU

Bloodborne Pathogen Program.

3. Poliovirus

All laboratory personnel who work directly with the agent are required to have documented polio

vaccination or demonstrated serologic evidence of immunity to all three poliovirus types.

4. Poxviruses: vaccinia, cow pox, monkey pox

Vaccination is required for those who work with or around orthopox viruses.

5. Rabies virus

Immunization is recommended for all individuals prior to work with rabies virus or infected

animals, or engaging in diagnostic, production, or research activities with rabies virus.

Immunization is also recommended for all individuals entering or working in the same room

where rabies virus or infected animals are used. The NAU Animal Care Program requires

immunization for all employees (e.g., veterinarians and animal control personnel) who come into

contact with unvaccinated carnivores.

6. Arboviruses

For the following arthropod-borne viruses there is a vaccine available and it is recommended for

those who work with the virus.

Eastern equine encephalitis Junin

Western equine encephalitis Mucambo

Cabassue Rift Valley Fever

Chikungunya Venezualan equine encephalitis

Everglades Yellow Fever

7. Measles, mumps, and German measles virus (rubeola, paramyxovirus, and rubella virus)

Immunizations are usually given in childhood. Vaccines are highly recommended for all

unvaccinated or serologically negative adults.

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4.1.2 Health Surveillance for Personnel Working with Infectious Agents

A health surveillance program is available for laboratory personnel who use agents that require it.

Laboratory personnel should receive immunizations (see chapter entitled “Immunoprophylaxis”), such as

hepatitis B vaccination, and medical tests, such as tuberculosis skin testing, when appropriate. The

Animal Care and Bloodborne Pathogen Programs (see appropriate chapters) provide for health

assessments, risk assessments, medical tests, and immunizations for certain at-risk personnel.

4.1.3 Blood Serum Sampling

For those laboratory personnel working with certain agents under BSL-2 or BSL-3, baseline serum

samples may be appropriate (see chapter entitled “Biological Safety Levels”). The collection and

maintenance of baseline serum samples provide a tool for monitoring serological changes that may result

from the employee’s work. For example, staff who work with non-human primates may be required to

bank a blood sample.

The EH&S Office of Biological Safety, with advice from the appropriate health care professional, will

provide information regarding the need for, and the frequency of, blood serum sampling on a case-by-

case basis. The need for serum sampling will be dependent upon the agent used and the type of research

in which an individual is engaged. For example, researchers who work with large quantities of HIV are

required to have serum sampling monitored annually and upon accidental exposure.

4.1.4 Health Assessments

The NAU Occupational Health Program provides for pre-placement and other health assessments depending upon

the type of work in which an employee will be engaged.

Laboratory personnel who work with agents that are transmitted by aerosol or with certain chemicals or

acute toxins may require a respirator at some stage of the research project. The EH&S Office of

Biological Safety Respiratory Protection Program provides for health assessments that will determine an

employee’s fitness for respirator use.

The EH&S Office of Biological Safety and Occupational Health Program also includes the Hearing

Conservation Program, the Asbestos Monitoring Program, as well as the Animal Care Program,

discussed in a separate section. Occasionally, researchers who work with infectious agents will require a

health assessment based on the requirements of one of these other programs.

4.1.5 Exposure to Mycobacterium Tuberculosis

All personnel who can be occupationally exposed to either TB research or patients with active TB should

have a yearly TB test for health surveillance.

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