Biological Safety - Laboratory Safety Training Sessions

Introduction

Proper care and use of laboratory animals is not only the humane thing to do, it is the law. Together, Duke’s Institutional Animal Care and Use Committee (IACUC) and Division of Laboratory Animal Resources (DLAR), maintain a program which assures compliance with the Animal Welfare Regulations and the Public Health Service Policy on Humane Care and Use of Laboratory Animals. For information about regulatory compliance or to request a copy of the Duke University Manual for Animal Research, contact the DLAR at 684-6745.

Employees working with animals in a research environment may be exposed to a variety of occupational risks including animal bites and scratches, needle sticks, back injuries from lifting large items, exposure to zoonotic agents, allergens, or hazardous chemicals, etc. In order to prevent accidents or exposures from occurring, all employees must understand the risks involved with such work, and must also be familiar with basic safety precautions for work with animals.

All proposed work involving animals must first be reviewed and approved by Duke’s IACUC before work can begin. The IACUC's homepage is found at: http://vetmed.duhs.duke.edu/

All orders for vertebrate animals must be placed by the DLAR. Purchase orders will be accepted only if accompanied by an approved Animal Use Protocol number.

All employees who work with animals are required to complete the Placement Health Review for Animal Handlers. This form will be reviewed by Employee Occupational Health and Wellness, which administers all applicable medical surveillance. For more information, contact EOHW at 684-3136.

Duke’s IACUC provides animal care and use training sessions which are mandatory for all animals handlers. The two required sessions are available online. These trainings provide detailed instruction on both regulatory compliance and basic husbandry. Because hazards vary from lab to lab, it is also necessary for each principal investigator (PI) to provide site-specific instructions for conducting animal work safely. This would include information on any etiologic agents, chemical hazards, radiation hazards, behavioral concerns of a particular animal species, etc. In addition to identifying the risks, the PI is also responsible for providing appropriate personal protective equipment such as lab coats, disposable gloves, goggles and face shields.

Husbandry

Animals shall be provided nutritionally adequate food, potable, non-contaminated water and a sanitary environment in which the animal’s health shall not be affected.

All sharps shall be disposed of immediately after use in an approved sharps container. Needles shall not be broken, bent, or recapped before disposal. A one-handed recapping procedure may be incorporated only after approval by the OESO-Biological Safety Division. Physical restraint procedures should be developed and practiced to prevent accidental autoinoculation while at the same time reducing stress on the animal.

Animal Restraint

Proper restraint and handling techniques are essential for reducing stress to laboratory animals, while at the same time allow animal care workers to perform their work with less chance of being scratched, bitten, kicked, etc. Animals can be restrained either manually or with restraint devices. It is the responsibility of the PI to train their staff on proper restraint for each species used.

Precautions should be taken, while changing animal bedding, to minimize or eliminate the aerosolization
of hazardous agents which may have been shed by the animal. Many allergens can also be aerosolized during bedding changing. Some options include the use of a cage changing hood, or decontaminating the soiled bedding before disposal by placing the whole cage in an biohazard bag and then autoclaving it.

One of the most common conditions that affects individuals who work with laboratory animals is allergies. Typically, allergies to animals are IgE mediated reactions, and result from exposure to an animal’s dander, urine, saliva, serum, or other body tissues. Symptoms can range from mild (i.e. itchy or runny nose and eyes) to severe (i.e. shortness of breath or red, itchy wheals on skin).

Levels of airborne allergens tend to rise significantly with certain activities such as changing or cleaning animal cages. The use of ventilated hoods for cage changing, dust-free bedding, or filtered caging systems, are all good ways of reducing the level of airborne allergens. If these options aren’t available or feasible for a particular situation, then personal respiratory protection may be warranted. Contact the OESO-Biological Safety Division at 684-8822 for more information.

All animal carcasses, which are non-radioactive, are to be collected by the DLAR staff and incinerated. For more information, contact the DLAR at 684-5567. Radioactive carcasses shall be segregated by half-life categories and labeled using the following color-coding scheme: (Green: half-life <30 days, i.e. 32P, 33P, 131I, and 51Cr; Yellow: half-life between 30 and 115 days, i.e. 125I, 35S, and 113Sn; Blue: half-life >115 days, i.e. 3H, 14C, 45Ca, and 211At) Radioactive carcasses and their associated lab waste (i.e. bedding, excreta, sharps, etc.) are to be bagged and sealed in 3 mil plastic bags. Make sure that all sharps are contained in a puncture resistant container before placing in plastic bags! Once properly tagged and labeled, the bags are refrigerated in a lined 30 gallon plastic drum. All packaging and labeling materials are provided the OESO. For more information contact the OESO- Environmental Programs Division at 684-2794.

Working with Hazardous Materials

A standard operating procedure must be developed and approved by OESO for any work which involves the the use of hazardous materials in animals. Such procedures shall detail the safe handling of the animal throughout the duration of exposure.

Work involving the exposure of animals with biological materials must be conducted at the appropriate containment level to ensure adequate protection of personnel and the environment. The following table summarizes the Center for Disease Control and Prevention's four animal biosafety levels.

ABSL	Agents	Practices	Safety Equipment (Primary Barriers)	Facilities (Secondary Barriers)
1	Not known to cause disease in healthy adults	Standard animal care and management practices, including appropriate medical surveillance programs	As required for care of each species	Standard animal facility non-recirculation of exhaust air directional air flow recommended
2	Associated with human disease. Hazard: autoinoculation, ingestion, mucous membrane exposure	ABL-1 practices plus: limited access biohazard warning signs sharp precautions biosafety manual decontamination of all infectious wastes and of animal cages prior to washing	ABL-1 equipment primary barriers: containment equipment appropriate for species: PPE: laboratory coats, gloves, face and respiratory protection as needed	ABL-1 facility plus: autoclave available handwashing sink available in the animal room
3	Indigenous or exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences	ABL-2 practices plus: controlled access decontamination of clothing before laundering cages decontaminated before bedding removed disinfectant foot bath as needed	ABL-2 equipment plus: containment equipment for housing animals and cage dumping activities Class I or II biosafety cabinets available for manipulative procedures (inoculation, necropsy) that may create infectious aerosols. PPE: appropriate respiratory protection	ABL-2 facility plus: physical separation from access corridors self-closing, double door access sealed penetrators 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	ABL-3 practices plus: entrances through change room where personal clothing is removed and laboratory clothing is put on; shower on exiting all wastes are decontaminated before removal from facility	ABL-3 equipment plus: maximun containment equipment (i.e.Class III biosafety cabinet or partial containment equipment in combination with full body, air supplied positive-pressure personnel suit) used for all procedures and activities	ABL-3 facility plus: separate building or isolated zone dedicated supply/exhaust, vacuum and decontamination systems other requirements outlined in the complete description of ABL-4
Summarized from Biosafety in Microbiological and Biomedical Laboratories, 4th Edition, 1999. http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm

Many anesthetic gases are used at Duke University for performing animal surgeries. These gases can present a risk for potential exposure to the lab personnel performing the surgeries. Anesthetics of concern include ether, nitrous oxide, and halogenated agents (i.e. halothane, isoflurane, methoxyflurane). Some of these halogenated anesthetics have been linked to adverse health effects in exposed workers, such as reproductive and neurological effects.

Emphasis must be placed on protecting personnel from exposure by adequately “capturing” the waste gas being generated. This may be accomplished by several methods depending on the method of delivery of the gas.

· perform work in a fume hood so when lid is removed, gases as captured by hood

· filter canisters must be maintained according to manufacturer’s specifications

· choose appropriate sized face-piece to ensure most efficient waste gas recovery

Lab personnel that are concerned with possible exposure to anesthetic gases may contact the Hygiene & Safety division of OESO at 684-5996 to request an exposure risk evaluation.

References
Guide for the Care and Use of Laboratory Animals; NRC, 1996
Guidelines for Research Involving Recombinant DNA; NIH, 2000
Biosafety in Microbiological and Biomedical Laboratories, 4th Ed.; CDC/NIH, 1999