FinalNTP_ReproSpecsMay2011.pdf

SPECIFICATIONS FOR THE CONDUCT OF STUDIES

TO EVALUATE THE REPRODUCTIVE AND DEVELOPMENTAL

TOXICITY OF CHEMICAL, BIOLOGICAL AND PHYSICAL AGENTS

IN LABORATORY ANIMALS

FOR THE NATIONAL TOXICOLOGY PROGRAM (NTP)

May, 2011

For test-article or study-specific questions in regard to these specifications, contact the Contract Officer’s Technical Representative i

TABLE OF CONTENTS

I. PERSONNEL ...................................................................................................................................... I-1 A. KEY PERSONNEL ......................................................................................................................... I-1 B. DISCIPLINE LEADERS ................................................................................................................. I-1 C. OTHER CRITICAL STAFF ............................................................................................................. I-4 D. SUPPORT STAFF ......................................................................................................................... I-5

II. FACILITY ........................................................................................................................................... II-1 A. FLOOR PLAN ............................................................................................................................... II-1 B. EMERGENCY FACILITY SUPPORT ............................................................................................ II-1 C. ANIMAL FACILITIES ..................................................................................................................... II-1 D. SUPPORTING FACILITIES .......................................................................................................... II-2

III. HEALTH AND SAFETY .................................................................................................................... III-1 A. ADMINISTRATIVE CONTROLS .................................................................................................. III-1 B. TEST ARTICLE/POSITIVE CONTROL HANDLING AND SAFETY POLICIES .......................... III-4 C. ENGINEERING CONTROLS ....................................................................................................... III-6 D. PERSONAL PROTECTIVE EQUIPMENT SELECTION ............................................................. III-9 E. FIRE SAFETY ............................................................................................................................ III-11 F. EMERGENCY PROCEDURE .................................................................................................... III-12 G. WASTE DISPOSAL/TEST ARTICLE SHIPMENT ..................................................................... III-12

IV. CHEMISTRY .................................................................................................................................... IV-1 A. GENERAL REQUIREMENTS ......................................................................................................IV-1 B. BULK TEST ARTICLE CHEMISTRY ...........................................................................................IV-1 C. DOSE FORMULATION ................................................................................................................IV-2 D. FORMULATION ANALYSIS ........................................................................................................IV-3 E. FORMULATION ANALYSIS REQUIREMENTS ..........................................................................IV-4 F. ANALYSIS OF DOSING VEHICLES ...........................................................................................IV-8 G. PRESTART CHEMISTRY REPORT ............................................................................................IV-8 H. BIOLOGICAL SAMPLE ANALYSIS .............................................................................................IV-8 I. DISPOSITION OF SURPLUS/RESIDUAL TEST ARTICLE ......................................................IV-10

V. LABORATORY ANIMAL MANAGEMENT AND TOXICOLOGY ...................................................... V-1 A. ANIMAL FACILITY OPERATIONAL REQUIREMENTS ...............................................................V-1 B. ANIMAL ROOM ENVIRONMENT .................................................................................................V-4 C. DIET AND WATER .......................................................................................................................V-5 D. CAGING, RACKS, BEDDING AND FILTERS...............................................................................V-7 E. ANIMALS ......................................................................................................................................V-9 F. ANIMAL DISEASE SCREENING PROGRAM ............................................................................V-11 G. SPECIAL REQUIREMENTS FOR GAVAGE ADMINISTRATION ..............................................V-14 H. REQUIREMENTS FOR STUDIES WITH A MATING COMPONENT ........................................V-14

VI. CLINICAL PATHOLOGY ................................................................................................................. VI-1 A. CLINICAL PATHOLOGY ASSESSMENTS .................................................................................VI-1 B. CLINICAL PATHOLOGY LABORATORY REQUIREMENTS ......................................................VI-3 C. REPORTING REQUIREMENTS ..................................................................................................VI-4

VII. PATHOLOGY .................................................................................................................................. VII-1 A. CAPABILITY ...............................................................................................................................VII-1 B. NECROPSY ................................................................................................................................VII-1 C. TISSUE FIXATION, TRIMMING AND STAINING.......................................................................VII-9 D. HISTOLOGY .............................................................................................................................VII-14 E. HISTOPATHOLOGIC EVALUATION ........................................................................................VII-16 F. RECORDING OF HISTOPATHOLOGY RESULTS ..................................................................VII-17 G. QUALITY CONTROL OF PATHOLOGY ACTIVITIES AND DATA ..........................................VII-18 H. SUBMISSION OF PATHOLOGY DATA ...................................................................................VII-19 ii

I. SUBMISSION OF HISTOPATHOLOGY MATERIALS .............................................................VII-20 J. RELEASE OF SLIDES ..............................................................................................................VII-22 K. REFERENCES ..........................................................................................................................VII-22

VIII. CAESAREAN-SECTION AND COLLECTION OF FETAL DATA .................................................. VIII-1 IX. QUALITY ASSURANCE .................................................................................................................. IX-1

A. GOOD LABORATORY PRACTICE (GLP) REQUIREMENTS ....................................................IX-1 B. THE QAU .....................................................................................................................................IX-1 C. AUDITS AND INSPECTIONS ......................................................................................................IX-2

X. GENERAL STUDY PROTOCOL OUTLINES ................................................................................... X-1 A. PROTOCOL OUTLINE FOR RACB STUDY ................................................................................X-2 B. PROTOCOL OUTLINE FOR PRENATAL DEVELOPMENTAL TOXICITY STUDY ...................X-11 C. PROTOCOL OUTLINE FOR THE NTP MODIFIED ONE GENERATION STUDY ....................X-15

XI. DATA COLLECTION AND SUBMISSION ....................................................................................... XI-1 A. DATA COLLECTION ....................................................................................................................XI-1 B. SUBMISSION OF REPORTS AND DATA ...................................................................................XI-1

XII. REPORT FORMATS ...................................................................................................................... XII-1 A. FORMAT AND GUIDELINES FOR MONTHLY PROGRESS REPORT ....................................XII-1 B. GENERIC STUDY REPORT FORMAT ......................................................................................XII-9

XIII. APPENDICES

APPENDIX 1 - Dose Analysis and Method Performance Evaluation APPENDIX 2 - Protocols for the Analysis of Dosing Vehicles Used in NTP Toxicity

Studies APPENDIX 3 - Laboratory Animal Management Appendix APPENDIX 4 - Individual Animal Necropsy Record APPENDIX 5 - Procedures for Sperm Motility & Counts and Vaginal Cytology

Evaluation (SMCVCE) APPENDIX 6 - Pathology Section A - Procedures for Collecting Non-Reproductive Organs Section B - Methods for Trimming Non-Reproductive Organs and Tissues Section C - Procedure for Ovarian Follicle Counts Section D - Tissue Collection Procedures for RNA, DNA and Protein Assays Section E - Preparation of Mammary Gland Whole Mounts in Mice and Rats APPENDIX 7 - General Information for Neurobehavioral Studies

I. PERSONNEL I-1

I. PERSONNEL

The Principal Investigator, Discipline Leaders for Toxicology, Chemistry, Pathology, Laboratory Animal Management, Health and Safety, Quality Assurance, as well as Study Directors, shall be employees of the testing laboratory (e.g., not consultants or subcontractors). The daily interaction and constant coordination of efforts needed amongst these discipline areas throughout the in-life portion of the studies makes it critical that they be physically and organizationally together.

A. KEY PERSONNEL

Principal Investigator - The Principal Investigator, as key personnel, shall be considered essential to the contract and shall be responsible for the daily management of the contract, point of contact for communication with the NTP Contract Officer’s Technical Representative (COTR, also referred to as Project Officer or PO), and in the communication between the NTP personnel and contract personnel. He/she shall be available to meet with NTP personnel during site visits.

Site visits by the NTP COTR involving NTP discipline leaders, and quality assurance audits of studies in progress, are routinely conducted at laboratories with ongoing studies. The Principal Investigator must be knowledgeable and up to date in all aspects of the Program/contract, including the status of toxicity studies, record keeping, reporting, and detailed analysis of cost for any or all functions which make up the total operation. General qualifications expected of a Principal Investigator include:

• A Doctorate, or the equivalent relevant experience in a key scientific discipline for toxicology testing (e.g., toxicology, physiology, pathology, veterinary medicine, biochemistry, developmental biology)

• Significant experience in managing large-scale rodent toxicology programs requiring coordination of all disciplines contributing to reproductive and developmental toxicology study programs.

• Demonstrated capability for effective communication, written and oral.

B. DISCIPLINE LEADERS

Individual(s) assigned to the following discipline roles are considered to be "critical staff" and must be capable of meeting the indicated requirements. The Discipline Leaders (DLs) are responsible for establishing scientific guidelines and procedures, training and supervision of professional and technical staff, and evaluation of results and performance within their discipline area relative to NTP requirements.

1. Discipline Leader for Reproductive and Developmental Toxicology

A Reproductive and Developmental Toxicologist shall be available to supervise and/or work with each Study Director as well as professional and technical toxicology staff. Experience in studying the mechanisms of action of toxic agents is desirable since this individual, in addition to contributing to the implementation of basic experimental designs, must be familiar with any supplementary protocols to be included in study of a particular agent. General qualifications of a DL for Reproductive and Developmental Toxicology include:

• A Ph.D. or an equivalent in the Biological Sciences with relevant formal course work in disciplines such as toxicology, embryology/developmental biology, anatomy, neurotoxicology, endocrinology, pharmacology, physiology, biochemistry, and related areas. A record of publications in peer reviewed journals, book chapters and/or demonstrated experience in preparation of relevant laboratory study reports. Board certification (American Board of Toxicology) is desirable.

• Experience in the conduct of developmental toxicity studies as well as multi-generational reproductive toxicity studies in laboratory animals.

I. PERSONNEL I-2

• Experience in the management of professional and technical toxicology staff.

2. Discipline Leader for General Toxicology

A General Toxicologist shall be available to supervise and/or work with each Study Director as well as professional and technical toxicology staff. Experience in studying the mechanisms of action of toxic agents is desirable since this individual, in addition to contributing to the implementation of basic experimental designs, must be familiar with any supplementary protocols to be included in study of a particular agent. General qualifications of a DL for General Toxicology include:

• A Ph.D. or an equivalent in the Biological Sciences with relevant formal course work in disciplines such as toxicology, pharmacology, physiology, biochemistry, and related areas. A record of publications in peer reviewed journals, book chapters and/or demonstrated experience in the preparation of relevant laboratory study reports. Board certification (American Board of Toxicology) is desirable.

• Experience in the conduct of a variety of toxicity studies including short and long-term toxicology studies and in the management of professional and technical toxicology staff.

• Experience with in vivo evaluation of toxicants in rodents of various ages (e.g., whole animal testing, treatment response) preferably in toxicology studies of both short and long-term duration.

3. Discipline Leader for Chemistry

A Chemist shall be available for supervision of, and/or work with, personnel performing chemical analyses, dose preparations, and dose analyses. General qualifications of a DL for Chemistry include:

• Ph.D. in chemistry with two years relevant experience or M.S. in chemistry with five years relevant experience.

• Experience in chemical analyses for purity and identity as well as dose mixing and dose analysis relative to those required in this Specifications plus experience in the use of analytical instrumentation including gas-liquid and high performance liquid chromatography, spectroscopy, instrumental and wet chemistry analyses.

4. Discipline Leader for Pathology

A veterinary or medical Pathologist with experience in laboratory animal rodent pathology shall be responsible for oversight of professional and technical pathology staff and all pathology procedures, histopathologic evaluations, and reporting. General qualifications of a Pathologist include:

• Formal training in a medical specialty, e.g., veterinary medicine or medicine.

• Experience in rodent pathology; particularly reproductive, developmental and toxicologic pathology.

• Experience in management/supervision of pathology projects and/or staff.

• Post-doctoral training and/or Board Certification in pathology.

5. Discipline Leader for Laboratory Animal Management A veterinarian, with experience in laboratory animal medicine, must closely monitor the health and maintenance of experimental animals, provide oversight for the vivarium, and interact

I. PERSONNEL I-3

with the Principal Investigator and the NTP personnel concerning all phases of the animal experiments. General qualifications of the DL for Laboratory Animal Management include:

• Graduate of Veterinary College recognized by the American Veterinary Medical

Association.

• Diplomate of the American College of Laboratory Animal Medicine (preferred) or eligible by experience and education to take the examination.

• Previous experience in managing large colonies of laboratory animals, in particular, rodents in a toxicology setting.

• Previous experience that required multidisciplinary interactions such as toxicology studies.

• Previous experience in laboratory animal management involving the conduct of studies using breeding, pregnant and lactating animals.

6. Health and Safety Officer A qualified Health and Safety Officer shall be designated to monitor worker health and safety conditions during all phases of the work. In his/her role as Health and Safety Officer, he/she shall be responsible to someone other than the Principal Investigator (PI) and the PI's subordinates, and shall have the authority to bring unsafe conditions to the attention of higher management. The Health and Safety Officer may have other responsibilities within the Testing Laboratory’s organization; however, the amount of time devoted explicitly to health and safety is to be commensurate with the scale of the Testing Laboratory's operations.

General qualifications of the Health and Safety Officer include:

• Bachelor's degree (at a minimum) majoring in industrial hygiene, chemistry, biology, safety engineering or a closely related science or engineering field.

• At least two years experience in occupational health and safety, along with completion of courses in general occupational health and hazard control indicating the acquisition of successively greater levels of knowledge regarding industrial hygiene. This experience shall have taken place within the last 4 years. (A Master's degree in industrial hygiene, safety engineering or a Bachelor's degree in industrial hygiene, safety engineering, with one year of experience, is an acceptable substitute for this experience.)

• Training shall have been completed within the last eighteen months, and will be refreshed with additional training at an interval not exceeding eighteen months.

• Recent experience in working with specific requirements of local, state, and federal statutes relating to occupational health and safety, environmental protection, and chemical monitoring.

• Demonstrated ability to deal effectively with the scientific and managerial staffs in responsibly implementing the health and safety program (including the identification of problem areas and the execution of corrective actions as required).

7. Quality Assurance Unit Officer

This individual shall be responsible for monitoring each study to assure management that the facilities, equipment, personnel, methods, practices, records, and controls are in conformance with the regulations in Part 58, "Good Laboratory Practices for Non-clinical Laboratory Studies (Federal Register, Friday December 22, 1978, Part II and any later interpretations published by the FDA). The Quality Assurance Unit Officer shall have

I. PERSONNEL I-4

management support and organizational independence from the personnel engaged in the direction and conduct of the study. General qualifications for a Quality Assurance Officer include:

• A minimum of a Bachelor's degree, majoring in a biological science or chemistry.

• Appropriate scientific experience to ensure understanding of the tasks or reports being inspected or audited in toxicology, chemistry, and histopathology.

• The QAU Officer should have supervisory experience when the unit consists of more than one person.

C. OTHER CRITICAL STAFF

1. Study Directors Individual(s) assigned as the GLP Study Director must have formal training in and experience with conducting studies according to regulations defined in Part 58, “Good Laboratory Practices for Non-clinical Laboratory Studies (Federal Register, Friday, December 22, 1978, Part 11 (and subsequent updates/interpretations published by the FDA). The Study Director is expected to have day-to-day interactions with technical staff, observe key in-life/necropsy phases, and must be available to observe all toxicological animal room activities. The Study Director must be capable of meeting the following requirements:

• A Ph.D. or an equivalent in the Biological Sciences with relevant formal course work (or equivalent) in disciplines such as toxicology, pharmacology, developmental biology, endocrinology, physiology, biochemistry, and related areas. Board certification (American Board of Toxicology) is desirable.

• Experience with in vivo evaluation of toxicants in studies with laboratory animals of all ages including reproductive and developmental toxicology studies (e.g. rodent reproductive, developmental and/or multigenerational studies) by various routes of exposure. Additional experience with general toxicity studies is desirable.

• Demonstrated capability for effective communication, especially written.

2. Pathologists Veterinary or medical Pathologists with experience in laboratory animal rodent pathology are responsible for the necropsy/histopathologic evaluation and reporting of a study. They must be available to readily respond to necropsy, histology, and other related issues. General qualifications of a Pathologist include:

• Formal training in a medical specialty, e.g., veterinary medicine or medicine.

• Experience in rodent pathology including gross and microscopic pathology of the reproductive system, and toxicologic pathology.

• Post-doctoral training and/or Board certification (American College of Veterinary

Pathologists).

3. Breeding Specialist Breeding Specialist with experience in the breeding of laboratory animals, particularly rodents such as rats and mice. General qualifications include:

• Knowledge of reproductive biology gained through education and experience.

I. PERSONNEL I-5

• Understanding of mating and maternal behavior, as it applies to the production of offspring for research purposes including toxicology studies.

4. Data Management Coordinator

Individual responsible for data management activities involving data collection, management, reporting, and archiving.

• Individual shall have a documented record of experience and training in this area, specifically as it applies to reproductive and developmental toxicity studies.

D. SUPPORT STAFF

1. Dose Formulation and Analysis Staff A dose formulation supervisor with experience related to the testing program. Appropriately trained technical staff to perform dose formulation, and bulk test article and dose formulation analysis.

2. Animal Care/Toxicology Staff Animal care/toxicology technicians with training and experience in the administration of test articles via the required routes of exposure (including direct dosing of neonates), in providing appropriate animal care, in performing standard reproductive assessments (including observations and activities related to littering parameters, sexing neonates and measurement of ano-genital distance), assessment of development markers of sexual maturation (i.e.

vaginal opening and preputial separation), performing neurobehavior assessments, sperm motility and vaginal cytology evaluations, and in evaluating clinical signs of toxicity in pregnant, neonatal and adult animals.

3. Necropsy and Histology Staff Prosectors trained and experienced in the anatomy and dissection of rodents (including detailed dissection of reproductive tissues), experienced in the conduct of fetal examinations (external, visceral and skeletal), able to recognize abnormal anatomic organs of neonatal and adult animals and to describe gross abnormalities, including those of involving the reproductive tract. ASCP (HT) or ASCP (MT) registered technologist to supervise the histology operations. Appropriately trained histology technicians.

4. Clinical Pathology Staff Registered technologist to supervise clinical laboratory studies. Appropriately trained personnel with experience conducting clinical laboratory tests including hormone analyses.

5. Quality Assurance Staff Appropriately trained staff responsible for conducting audits and inspections of study activities.

6. Data Management Staff Individual(s) responsible for data management activities involving data collection, management, reporting, and archiving aspects.

I. PERSONNEL I-6

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II. FACILITY II-1

II. FACILITY

A. FLOOR PLAN

Laboratories conducting studies for the NTP shall provide floor plans showing locations where all study activities are conducted.

1. Animal Facility

Floor plan indicating quarantine rooms, animal rooms, showers/change areas and rest rooms; storage areas for feed, bedding and general storage; cage and rack washers;

emergency power source and those areas in which emergency power operates. This floor plan is to indicate traffic flow for personnel, animals, test article, feed/bedding, supplies, and equipment through the facility.

A separate floor plan of the animal facility is to indicate room airflow directionality and indicate the location of all safety equipment, such as eyewash stations, safety showers, fire control equipment, etc.

A floor plan indicating ventilation equipment and ductwork, including interior and exterior exhausts, shall be supplied. A floor plan for the roof(s) shall indicate the location of each of the building(s) general air intakes and exhausts and the location of the exhaust for each hood or vented enclosure.

2. Chemistry Indicate storage areas for bulk chemical and dose formulations, bulk chemical and dose formulation analysis, dose formulation, and supporting equipment and exhaust hoods.

3. Clinical pathology Indicate laboratory space and location where specimens are collected and analyzed.

4. Fetal examinations Indicate laboratory areas where fetal examinations and processing take place.

5. Pathology

Indicate areas for necropsy, histology, pathology, and storage.

6. Other areas Indicate QA offices and filing space; data archives; waste storage; and special study facilities.

B. EMERGENCY FACILITY SUPPORT

Facilities shall have a tested back-up power source with automatic changeover equipment that is sufficient to preserve the integrity of the testing experiment. Emergency power must handle those areas critical to the study such as animal rooms, HVAC, storage freezers/refrigerators, waste storage, autotechnicons, etc. Essential mechanical equipment must be guarded or alarmed. Provisions for prompt maintenance response must be provided.

C. ANIMAL FACILITIES

All facilities for the Testing Program must be approved by the NTP and will be evaluated with respect to criteria outlined in the "Guidelines for Carcinogen Bioassay in Small Rodents" (DHHS Publication No. (NIH) 76-801), "Long Term Holding of Laboratory Rodents" (ILAR News, XIX, #4, 1976), the "Guide for the Care and Use of Laboratory Animals for Research Involving Chemical Carcinogens" (DHHS Publication No. (NIH) 76-900), the "Guide for the Care and Use of Laboratory Animals NRC, 2010), and any other additions and exceptions thereof. Research is to

II. FACILITY II-2

be conducted in accordance with the Public Health Services Policy on Humane Care and Use of Laboratory Animals, Office of Laboratory Animal Welfare (OLAW). Accreditation of toxicology research and testing facilities by an external peer review organization such as AAALAC (Association for Assessment and Accreditation of Laboratory Animal Care International) or CCAC (Canadian Council on Animal Care) is required. It is the responsibility of the testing laboratory to establish policy and procedures to address entry of approved staff and visitors into the animal facility. Entry shall be prohibited to those individuals that have been in another animal facility within the last 48 hours regardless of disease status of that facility. The animal facilities shall be designed and managed to prevent contamination of animals with pathogenic organisms and to prevent contamination of personnel and the environment with test articles and also to prevent cross-contamination of animals with other test articles. A two-corridor system with intervening animal rooms is the preferred way to fulfill this requirement, given that:

1. All materials coming in contact with animals are sanitized to a clean state suitable for introduction to the supply (clean) corridor and the animal rooms.

2. Following use, these materials are removed from animal rooms by a return (dirty) corridor for disposal, destruction, or reprocessing.

3. The air pressure is adjusted so that the animal rooms are positive to the return corridor and negative to the "clean" one.

4. In cases where animals must be moved from one room to another, such as in multigeneration studies or conduct of neurobehavior tests, precautions must be followed so that the barrier system is not compromised.

D. SUPPORTING FACILITIES

1. Animal Facility

a. Facilities for sanitization of cages, racks, water bottles, and feeders must be available and properly located in relation to the study rooms.

b. Clean, well-ventilated, vermin-free storage space must be provided for clean supplies and equipment, cage filters, and feed/bedding awaiting use.

c. Ability to autoclave feed and bedding is desirable.

2. Pathology

a. The necropsy facility must be equipped with adequate working surfaces, dissection boards, running water with drains, dissection microscopes, perfusion equipment, adequate lighting, ventilation, and exhaust hoods. Necropsy and microscopic photography capabilities are required. The location of pathologists’ offices shall allow them to be readily available to address necropsy issues and attend necropsies.

b. Refrigeration shall be available for holding dead animals until necropsy. Dead animals shall not be frozen prior to necropsy.

c. The histology laboratory should be separated from the necropsy area and equipped with automatic tissue processor(s), microtomes, embedding and staining equipment, and with supplies and appropriate ventilation adequate for the expected volume.

d. Acceptable storage space must be available for storage of residual archival and histologic materials that will be retained by the Testing Laboratory prior to shipment to the NTP-designated recipient. These areas must have secured, limited access.

II. FACILITY II-3

3. Chemistry and Hazardous Waste Storage

1. The contractor must have the analytical instrumentation required for dose analysis and chemical purity checks. The specific types of equipment required will depend on the test article(s), but might include gas and/or liquid chromatographs, infrared spectrophotometers, UV-visible spectrophotometers, and other similar instruments and apparatus.

2. Secured, controlled access bulk storage facilities must be available for retention of the test article at 5º C ± 3º C or at ambient temperature as specified by the NTP. Also, cold storage at –20º C ± 5º C must be available for analytical reference standards.

3. Suitable V-blenders with intensifier bars must be used for diet mixing.

4. Designated areas shall be provided and described for hazardous waste storage prior to disposal.

4. Data and Quality Assurance Archives

A secured, limited access area shall be provided for maintenance of all study records. In addition, Quality Assurance records must also be stored in a secured, limited access area.

II. FACILITY II-4

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III. HEALTH AND SAFETY III-1

III. HEALTH AND SAFETY

A. ADMINISTRATIVE CONTROLS

1. Regulations and Guidelines The NTP and/or its representatives may inspect, photograph, sample and monitor the laboratory and associated facilities used for its studies at any time to ensure that NTP minimum requirements and applicable regulations and guidelines (described below) are being followed. Any deviations to these requirements shall be approved by the NTP.

All work shall conform to applicable local, state, and federal statutes including the following federal regulations and updates:

OSHA:

• Standards for General Industry, 29 CFR 1910

• Hazard Communication, 29 CFR 1910.1200

• Respiratory Protection, 29 CFR 1910.134

• Occupational Exposure to Hazardous Chemicals in Laboratories, 29 CFR 1910.1450

• Occupational Exposure to Blood-borne Pathogens, 29 CFR 1910.1030

• Formaldehyde, 29 CFR 1910.1048 (applicable to the use of formaldehyde in histology, pathology, and anatomy laboratories)

DOJ:

• Americans for Disability Act, Accessibility, Design Guidelines, 28 CFR, Title III, Part 36.

EPA:

• Clean Air Act, 40 CFR 50-80

• Clean Water Act, 40 CFR 100-140 and 400-470

• Resource Conservation and Recovery Act (RCRA) 40 CFR 240-271

• Comprehensive Environmental Response, Compensation and Liability Act (CERCLA, Superfund, SARA) 40 CFR 300.

DOT:

• General Information, Regulations, and Definitions, 49 CFR 171

• Hazardous Material Table, Special Provisions, Hazardous Materials Communication

Requirements and Emergency Response Information Requirements, 49 CFR 172

• Shippers, General Requirements for Shipments and Packaging, 49 CFR 173

• Carriage by Public Highway, 49 CFR 177

NRC:

• Standards for Protection against Radiation, 10 CFR 20

• Notices, Instruction, and Reports to Workers; Inspections, 10 CFR 19

• Recommendations described in the most recent version of the NIH Radiation Safety

Guide

DEA:

• Federal Requirements for Controlled Substance, 21 CFR 1300

For contract work involving infectious agents, the Centers for Disease Control Guidelines, Biosafety in Microbiological and Biomedical Laboratories (HHS Publication No. (NIH) 93- 8395, 2009) and the NIH Guidelines for Research Involving Recombinant DNA Molecules (66 Federal Register 57970, 2001 and updates) shall be followed.

III. HEALTH AND SAFETY III-2

Where not superseded by this document, the American National Standard for Laboratory Ventilation, Z 9.5, published by the American National Standards Institute (ANSI) shall be followed.

Other consensus standards and publications may include: the current edition of the Threshold Limit Values for Chemical Substances and Physical Agents & Biological Exposure Indices published by the American Conference of Governmental Industrial Hygienists (ACGIH), Criteria Documents for various substances and the Recommended Exposure Limits published by the National Institute for Occupational Safety and Health (NIOSH), and the Workplace Environmental Exposure Levels (WEEL) published by the American Industrial Hygiene Association. Where there may be conflict in the acceptable exposure levels as compared to the OSHA Permissible Exposure Limit, the most stringent standard shall be used for worker’s protection.

2. Health and Safety Plan (Chemical Hygiene Plan)

a) Health and Safety Plan Scope

The scope of each Health and Safety Plan shall address the organization's health and safety policies, Occupational Medical Surveillance Program, as well as potential chemical, physical, biological and ergonomic hazards (e.g., acquisition of study materials, storage, and handling through ultimate disposal of contaminated wastes).

No contract laboratory will participate in studies without a Health and Safety Plan that has been approved by the NTP. An updated Plan shall be submitted every 2 years to the NTP for review. In addition, the NTP shall be informed of any updates to the Plan during the course of the contract. If approval of the Plan is not granted at the time of award, the laboratory must submit a revised Plan for review within 30 days of the receipt of award notification. Revisions to the Plan shall be clearly indicated to facilitate reviewer approval.

For all contract laboratories, a Chemical Hygiene Plan as required under the OSHA "Laboratory Standard" may be used in place of a Health and Safety Plan provided it meets or exceeds ALL of the requirements outlined in this document.

b) Health and Safety Plan Content

Written Policies. In addition to the SOPs outlined below, the Health and Safety Plan shall address (but not be limited to):

• health and safety responsibilities, policies and organization

• record keeping and archiving

• initial and periodic employee training

• engineering controls

• personal and environmental monitoring

• medical surveillance and biological monitoring

• respiratory protection program

• personal protective clothing and equipment

• general housekeeping

• eating and smoking policies and areas

• precautionary signs and labels

• chemical and biological storage

• fire protection and prevention

• emergency and evacuation contingencies

III. HEALTH AND SAFETY III-3

• locations (with schematic diagrams) of fire control equipment, and plumbed eyewash stations and emergency showers

• laboratory safety inspection

• waste management and disposal

• other pertinent personnel, operational, and administrative practices, and engineering controls necessary for the containment and safe handling of chemical, physical, biological, and radiological hazards.

• entry and exit to restricted areas

• visitors

3. Standard Operating Procedures

The laboratory shall be required to have Standard Operating Procedures (SOPs) that have been reviewed and approved by NTP for at least the following activities:

• visitors access to test areas

• employee training

• medical surveillance and biological monitoring

• respiratory protection, mask-fit, cleaning/maintenance, and inspection

• eye and face protection

• personal protective clothing and equipment

• general housekeeping practices

• ventilation system maintenance

• storage, receipt, transport, and shipping of study materials

• hazardous material handling (e.g., in analytical chemistry labs)

• dose preparation (if applicable)

• entry and exit from the limited access areas (including traffic patterns of dose prep facility and animal handling and testing room) (if applicable)

• spill clean-up, accident, emergency response and evacuation (including natural disasters) and fires/explosions

• use of radio-labeled material, infectious agents, and/or controlled substances (if applicable)

• waste management and disposal

4. Exposure Evaluation and Control

a) Permissible Exposure Limits/OSHA-Regulated Substances

All laboratories shall ensure that employees' exposures to hazardous substances do not exceed the permissible exposure limits (PELs) specified by OSHA in 29 CFR 1910, subpart Z. In addition, initial monitoring of employees' exposure to any substance regulated by a standard (29 CFR 1910.1001-1101) which requires monitoring shall be conducted if there is reason to believe that exposure levels for that substance routinely exceed the action level (or in the absence of an action level, the PEL). If this initial monitoring reveals that an employees’ exposure exceeds the action level or the PEL, the exposure monitoring provisions of the relevant standard shall be complied with.

If a PEL has not yet been established for a study material, alternative acceptable exposure standards (e.g., TLV®, REL, WEEL) shall be used (Refer to Section III.A.1.). In situations where there is no known exposure standard for a proposed test article, a suitable interim exposure standard based on current toxicology and industrial hygiene literature shall be established when feasible.

b) Formaldehyde Monitoring in Histology, Pathology, and Anatomy Laboratories

III. HEALTH AND SAFETY III-4

Histology, pathology, and anatomy laboratories must comply with the formaldehyde OSHA standard, 29 CFR 1910.1048. The use of formaldehyde in all other laboratories shall be carried out in accordance with the OSHA formaldehyde standard.

Histology, necropsy, tissue storage, and tissue trimming operations shall be conducted in a manner that employs engineering controls to ensure that airborne concentrations of formaldehyde do not exceed 0.75 ppm as an 8-hour TWA or 2 ppm as a 15-minute STEL. Monitoring shall be performed to evaluate exposure levels (both TWA and STEL) of workers potentially exposed to formaldehyde hazards. If the results of initial monitoring indicate exposure levels exceeding either 0.5 ppm as an 8-hour TWA (action level) or 2 ppm as a 15-minute STEL, additional monitoring must be performed. If the TWA exceeds the action level, sampling must be repeated every 6 months. If the STEL exceeds 2 ppm, sampling must be repeated annually. The repeat sampling may be discontinued when 2 consecutive sampling rounds are below the STEL and action level as described in the OSHA formaldehyde regulation, 29 CFR 1910.1048. In addition, all other provisions of the OSHA formaldehyde standard must be adhered to.

c) Test article/Positive Control Monitoring

Exposure monitoring shall be routinely conducted where both test article and controls are handled and when the test article/positive control has an established exposure standard such as the PEL, TLV®, REL, or WEEL of 10 ppm or less, or 0.1 mg/m3 or less. This exposure monitoring shall be performed at least once during initial dose preparation, once during initial dose administration, and at the midpoint of the study or every six months. Where there is no known exposure standard for a proposed test article, the contractor shall perform exposure monitoring at the same frequency stated above.

Determination of exposure and adoption of controls shall be based on a pre-determined interim exposure standard, when feasible.

5. Occupational Medical Surveillance

An occupational medical surveillance program is to be implemented to cover personnel who will be working with study test article or animals. The frequency of the surveillance as well as the scope of the medical examination shall be specified in the laboratory’s Health and Safety Plan. Persons who are required to wear respirators must obtain written medical clearance from an occupational health service provider for use of this equipment.

6. Injury and Incident Reports

A record shall be kept of all injuries or illnesses, including animal bites. In addition any record of an OSHA recordable incident shall include a full description of the incident, the test article/positive control involved, the medical attention required, any remedial actions taken, and planned follow-up to minimize the likelihood, or eliminate the potential for, reoccurrence (if pertinent). Copies of such incident reports shall be forwarded to NTP.

The Project Officer shall be notified IMMEDIATELY if a serious (as defined by OSHA) accident or incident occurs.

All occupational injuries and illnesses shall be recorded and reported according to the OSHA recording system.

B. TEST ARTICLE/POSITIVE CONTROL HANDLING AND SAFETY POLICIES

1. Receipt/Handling/Storage

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A log shall be maintained which will include the date of test article receipt and a continuous balance of the remaining amount of test article.

Weighing of the test article and/or positive control shall be done using the smallest quantity needed. An analytical balance shall be used whenever possible to preclude the need for handling large amounts of chemical. This balance shall be placed at all times in an effective laboratory hood or a vented enclosure exhausted to the outside (see Section III.C.3.).

Protocols shall be designed to use the minimum possible quantities of "neat" chemical in preparing solutions.

A non-breakable, secured secondary container shall be used for transfer of any test article and/or positive control.

Volatile test articles shall be handled properly (e.g., keeping lids on container when not in use, segregating from unintended contact with heat or high pressure, etc.) and stored in an area with adequate ventilation that is directly vented to the outside. All other test articles shall be stored in a secured, designated storage area(s). However, flammable liquids must be stored in a non-vented flammable liquid storage cabinet (see Section III.E.1.).

2. Hazard Communication

a) Training

Personnel who handle (receive, store, weigh, dilute, transport, package, or administer) hazardous agents shall be provided with written material and trained on the associated hazards of these agents including the contents of the Material Safety Data Sheet (MSDS). This training shall be conducted by the HSO or a program approved by the HSO and shall be properly documented. Training shall include the recommendations for handling carcinogens. In addition, training in accordance with the requirements of applicable regulations shall be conducted.

b) Labeling

Warning signs and labels shall be used wherever test articles are used or stored (e.g., on primary and secondary containers, affixed to entrances to work areas, refrigerators, and on containers holding hazardous waste). These signs and labels shall be conspicuous (especially for containers to minimize handling) and shall indicate the presence of suspected carcinogenic, mutagenic, and other hazards, as required by OSHA.

c) Health and Safety Documents

The contractor must have available for each study agent and positive control health and safety documentation that includes, but is not limited to, the supplier’s MSDS*, which includes information on the material's hazards, properties and appropriate control measures. All employees handling the study material and/or positive control must be trained regarding the contents of the agent-specific health and safety data document.

MSDS shall be accessible at all times at designated locations known by the employees.

• If a chemical is produced for a user outside of the laboratory then the laboratory is required to develop an MSDS.

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C. ENGINEERING CONTROLS

1. General Facility Requirements

Safety showers, drench hoses, and eyewash stations shall be located throughout the facility as required by local, state, and federal regulations and must be located in close proximity to where potentially hazardous chemicals are stored or used. Only plumbed eyewashes are permitted.

2. Isolation and Access Restriction

a) General Requirements for Access Restriction

An isolated, posted, restricted access laboratory (or laboratories) separate from other laboratory facilities shall be designated for unpacking, storing, weighing, and diluting of test articles and/or positive controls; and where necropsy, tissue trimming, tissue processing, embedding, microtoming and staining are performed.

Administration of test article and positive controls shall be performed in a limited access area with its air supply under negative pressure with respect to connecting laboratories and hallways. This shall be a separate laboratory from the area described above for areas dedicated to unpacking, storing, weighing, and diluting.

Each lab shall have a room inspection program providing monthly checks of the air flow directionality. Relative pressures of laboratory areas shall be checked monthly with smoke tubes to verify that air flows from relatively clean to relatively dirty areas. Monthly inspections shall be documented.

A record shall be kept of all personnel entering/exiting any limited access area(s).

b) Requirements for Barrier Systems

The dose preparation area shall be isolated from general traffic. This may be accomplished by locating the dose preparation area within the animal facility limited access barrier system, or by establishing a separate limited access area for dose preparation. If the latter approach is used, all areas into which laboratory workers may bring used protective equipment (including gloves, shoes, head covers, and clothing), respirators, and/or containers of dosed feed or water shall be behind the barrier. Also, any hallways used by workers for reaching the shower facility shall be considered to be behind the barrier (e.g., limited access area).

Personnel who enter the dose preparation area, or an area requiring a complete set of clean protective clothing and equipment (e.g., a disposable laboratory suit, safety goggles, disposable gloves with permeation-resistant properties specific to the test article, disposable boots, disposable shoe covers or sneakers or rubber boots, and disposable head covering), must shower out prior to leaving the barrier facility at the end of the day.

Within the shower facility, the "clean" and "dirty" sides must be physically separated by the shower or by another physical barrier. The facility design and procedures shall be arranged so that it is not necessary to require entering the clean side prior to showering and to prevent returning to the dirty side after showering (e.g., to store or retrieve items such as shoes, towels, respirators, etc.).

Each laboratory shall have a room inspection program providing monthly checks and documentation of the air flow directionality. Relative pressures of laboratory areas shall

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be checked monthly with smoke tubes to verify that air flows from relatively clean to relatively dirty areas.

c) Facility Design for Barrier Systems

Air exhausted from dose preparation areas involving the particulate form of the test materials shall be passed through HEPA filters. If volatile chemicals are handled, charcoal filters shall also be used. These filtration systems shall be periodically monitored and maintained and personnel performing maintenance shall wear the protective clothing described for neat test article handling. (See Section III.D.1.a.)

The relative location of external air intakes and exhausts for both local and general ventilation systems must be arranged to minimize the risk of re-entrainment of exhaust air. Documentation (e.g., schematic diagram) shall be provided to NTP indicating the location of intakes and exhausts, stack height, discharge velocities, as well as the direction of prevailing winds. No weather caps or other obstructions shall be in the path of vertical discharge.