Animal BioSafety

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Animal Research

The use of live animals in research presents many additional risks to workers.

Institutional Oversight

Institutional Animal Care and Use Committee (IACUC)

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Institutional Animal Care and Use Committee (IACUC)

Comprised of researchers, veterinarian, biosafety experts, statisticians, community members, and other UVM administrative staff. The IACUC reviews all vertebrate animal use protocols at the University to ensure the humane care and use of animals for research and education under optimum conditions that also comply with animal welfare regulations. IACUC approval is needed before any animal work can begin. The IACUC also conducts semi-annual inspections of all University areas where research animals are housed or used.

University Veterinarian

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The first point of contact for the IACUC review process. Provides specialized guidance on proper methods, housing, anesthesia, and drugs for animal procedures and care.

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Institutional Biosafety Committee (IBC)

Comprised of researchers, biosafety officer, community members, and other UVM administrative staff. IBC approval is required when using biohazardous materials in animals. The IBC reviews all work with recombinant or synthetic nucleic acids, infectious agents, viral vectors, biotoxins, and other potential biohazards.

EHS and Risk Management

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Risk Management

Environmental Health and Safety

A risk assessment is used to identify the potential hazards of the research project, as well as the appropriate controls to minimize the risk. The risk assessment is conducted by Safety staff and serves as an important tool along with input from the PI, facility manager, veterinarian, IACUC, and IBC if applicable.

Facility Manager

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Responsible for oversight of animal care staff, keeping facility running smoothly and safely, and making sure work is being conducted in the appropriate areas with the appropriate controls.

Occupational Health Provider

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Screens individuals for health risks such as animal allergies, asthma, etc. based on the work that they will be doing. The OHP is also available to provide care and treatment to any employees or students that become ill or injured as a result of their work.

Risk Assessment

Risks

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A risk assessment is a valuable tool for identifying the potential hazards and establishing safe laboratory operations. It is conducted on a case-by-case basis and should include, but not be limited to:

  • Pathogenicity and virulence – How easily the agent can infect the host (mode of infection, is the agent attenuated, wild type, etc.)
  • Type of toxicity – Which systems or organs are targeted?
  • Route of exposure - Inhalation, ingestion, mucous membranes, dermal, or bloodborne?
  • Transmission – Can the agent be passed between animals, from animals to handler, from handler to animals?
  • Infectious dose or toxic dose – How many individual organisms does it take to cause disease? (Example: Cryptosporidium parvum = as few as 10, Salmonella typhimurium = 100,000) If using a chemical, what is the toxic dose?
  • Availability of pre- and post-exposure prophylaxis – Pre-exposure vaccines are available for some diseases, but not all. Some post-exposure treatments are available, but they must be administered within a certain timeframe after exposure to be effective
  • Severity of reaction – Can range from mild to acute. Will exposure cause illness, irritation, anaphylaxis, chronic disease, etc.?
  • Environmental Stability – How long the agent can remain infectious or toxic outside of the animal
  • Metabolism or Shedding of agent by host – Will it be excreted in feces or urine, and if so, at what level? 
  • Hazards associated with species or animals – Aggression, size, difficulty of handling/procedures, housing
  • Methods and Researcher proficiency – Are there any high-risk techniques being used? Do the personnel have experience with the specific work they are performing? Has everyone been properly trained?

Animal Biosafety Levels

ABSL-1

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Required PPE and appropriate biohazards signs should be posted in facility

  • Risk Group 1 agents, not known to cause disease in healthy adult humans.
  • No recirculation of exhaust air
  • Directional airflow recommended
  • Hand washing sink recommended
  • Required PPE: lab coats, gloves, and eye/face protection as needed.
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Required PPE and appropriate biohazards signs should be posted in facility

ABSL-1 practices, plus:

  • RG2 agents, associated with human disease that is rarely life-threatening and for which treatments are available
  • Limited access to facility
  • Biohazard warning signs posted
  • Safe sharps precautions implemented
  • Decontamination of infectious wastes, and of cages prior to washing (mechanical cage washer recommended)
  • Primary containment equipment used
  • Autoclave available
  • Negative airflow into facility rooms
  • Hand washing sink available in animal rooms
  • Required PPE: lab coats, gloves, eye/face and respiratory protections as needed

ABSL-3

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Required PPE and appropriate biohazards signs should be posted in facility

ABSL-2 practices, plus:

  • RG3 agents, may cause serious or potentially lethal disease for which treatments may not be available
  • Decontamination of clothing before laundering
  • Controlled access to facility
  • Cages decontaminated before bedding removed
  • Containment for cage dumping required
  • Biosafety cabinets used for animal manipulations
  • Physical separation from main corridor (entry through anteroom)
  • Negative airflow into all rooms
  • Sealed windows and all penetrations
  • Self-closing, interlocked doors
  • Required PPE: full body protective clothing, lab coats/gowns, double gloves, eye/face protection, respiratory protection

ABSL-4

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Required PPE and appropriate biohazards signs should be posted in facility

ABSL-3 practices, plus:

  • RG4 agents, high risk of aerosol transmission, known to cause infections that are frequently fatal, for which there are no vaccines or treatments available
  • Separate facility or isolation zone
  • Dedicated supply and exhaust, vacuum, and decontamination systems
  • Entry through locker room where personal clothing is changed for dedicated laboratory clothing
  • Maximum primary containment used (class III BSC or glove box)
  • All wastes decontaminated before removal from facility
  • Shower out before exiting facility
  • Required PPE: positive pressure, air-supplied, full-body suit

Training and Prerequisites

Occupational Health Screening

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A medical questionnaire must be completed and submitted to the University’s Occupational Health Care Provider for review by a medical professional before someone is cleared for working with animals. The questionnaire will help to screen for any elevated risks to the employee. A physical exam, testing or vaccination, or respirator use may be required based on the risks involved.

Training

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Prior to contact with animals, all personnel must be trained on proper handling techniques, euthanasia if applicable, entry/exit procedures for the facility and animal rooms, emergency action, proper PPE and donning/doffing procedures, and specific SOPs for the work that they will be performing. Species-specific training may also be required.

Pathogen Safety Data Sheets for the agents being used, as well as possible zoonoses, should also be available. Employees should familiarize themselves with the signs and symptoms of infection before starting work.

CITI online training is required of all personnel listed on IACUC protocols.

Personal Protective Equipment

PPE

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  • Gloves must always be worn for animal contact, cage change procedures, handling biological materials, or handling hazardous chemicals or drugs
  • Body protection such as barrier gowns, disposable lab coats, or jumpsuits prevent contamination of clothing
  • Eye and face protection should be worn if there is the risk of a splash, aerosol, or as determined by a risk assessment
  • Respirators may be required to protect against allergens or biohazardous aerosols – N95 (disposable) are the most commonly used. Personnel must be medically cleared and fit-tested before using any kind of respirator.
  • Booties must be worn to protect against contamination in the animal facility. Booties must be worn over lab-appropriate, closed-toed shoes.
  • Head covers provide additional protection against contamination for both you and the animals in the facility. Always keep long hair tied back.

Additional PPE such as aprons, sleeves, rubber boots, or cut/puncture-resistant fabrics may be required based on facility requirements or risk assessment.

More about PPE

Species-Specific Hazards

Mice and Rats

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  • 95% of all research animals
  • May be difficult to handle and manipulate due to small size, small sharp teeth can cause puncture wounds, mothers can be protective of pups, some strains are more evasive or aggressive (i.e. cotton rats)
  • Use restraint devices or anesthesia to reduce risk of bites and accidental needle sticks – note: these methods must also be approved by the IACUC
  • Most lab strains have been bred to be “pathogen-free”, but all animals still carry naturally occurring bacteria in their saliva and digestive systems. A scratch or bite may introduce these bacteria, or naturally occurring bacteria on your skin, into the wound and cause an infection.

Fish and Amphibians

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  • Increased risk for splashes
  • Zoonotic microorganisms are shed into tank water

Ferrets, Rabbits, Guinea Pigs

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  • Higher risk for scratches and bites – larger and stronger than rats and mice
  • Ferrets share several common parasites with dogs and cats

Birds

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  • Not generally supplied as “pathogen-free” – may be wild caught or come from established flocks
  • Infectious diseases may be carried without symptoms for long periods of time, and shedding may be triggered by stress (such as shipping or moving)
  • Increased risk or airborne particles due to flapping of wings

Cats and Dogs

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  • Infections from bites are common - Need specialized handler training to learn how to avoid bites and scratches
  • Toxoplasmosis is of particular concern to pregnant women and immunocompromised people

Wild Animals (field studies)

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  • Use extreme caution, as wild animals are unpredictable and may be carrying zoonotic diseases

Livestock

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  • More difficult to obtain “pathogen-free” animals since many animals come from local farms, and animals are often housed in open barn – biosecurity is a major concern – wild rodents, insects, birds, are all common in barns
  • Present additional hazards like kicks due to large size, more difficult to restrain
  • Increased risk of splashes, aerosols – primary containment is a lot more difficult with large animals

Non-human Primates (NHP)

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  • These animals are extremely intelligent, strong, agile, and often unpredictable. The jaw strength of a NHP is about 10 times that of a human. Be aware of their potential reach zone from the cage, and do not wear anything loose or dangly, and always tie back long hair. Eye and face protection is essential, since NHPs may spit or throw things.
  • Highest risk species for zoonotic diseases
  • Personnel should complete extensive specialized training before working with NHPs.
  • Note: There are no NHPs at UVM.

Allergies and Zoonotic Diseases

Laboratory Animal Allergy (LAA)

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  • 10% of animal workers with no previous allergies will develop one
  • 70% of animal workers with existing allergies will develop a new one
  • Allergies can be to animal hair or skin, but more often to saliva or urine - allergens become aerosolized in animal bedding just by the regular movement of the animals in the cage, making cage changing/dumping and even higher risk for exposure
  • Symptoms may be respiratory, mucous membrane irritation (watery, itchy eyes, stuffy or runny nose), dermal irritation (eczema, hives, etc.)
  • Allergies tend to get worse with repeated exposure
    • Can lead to asthma (20-30% of those with allergies) or anaphylaxis (life-threatening medical emergency)
    • Report symptoms to your supervisor
  • Those with pre-existing allergies should wear a respirator. Some without pre-existing allergies choose to wear a respirator to prevent development of allergies
  • Main source of allergen exposure is though inhalation
  • Dust masks or surgical masks do not provide adequate protection, must use a NIOSH-approved respirator for particulates
  • Other ways to prevent LAA:
    • Use filter-top cages
    • Only open cages in biosafety cabinet, or use specifically designed dumping stations for cage dumping
    • Wear proper PPE – skin protection, eye/face protection
    • Animal care staff wear scrubs, change before going home
    • Facility design – negative pressure rooms, 10+ room air changes per hour
    • Good work practices and hygiene – glove changing, hand washing, general housekeeping

Zoonotic Diseases

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Zoonotic diseases, or zoonoses, are diseases that can be transmitted between humans and animals. Be aware of the potential zoonotic diseases for the species that you are working with and take precautions to avoid transmission. The following are examples, and not an exhaustive list.

  • Aeromonas - fish
  • Bartonella (Cat Scratch Disease) - cats
  • Brucellosis – cattle, pigs, goats, sheep, dogs (rare)
  • Campylobacter – fish, amphibians, birds, dogs, pigs
  • Cercopithecine Herpes Virus-1 (B Virus) - NHP
    • High prevalence in some captive species (macaques) - they carry this virus naturally with mild or no symptoms. Exposure to B Virus caused the death of a Yerkes worker in 1997. The mortality rate in humans is high. The virus can survive for up to 7 days on contaminated surfaces and can be transmitted through contact with skin and mucous membranes.
  • Chlamydia psittaci - birds
  • Coxiella burnetti (Q-Fever) – sheep, goats, cattle
  • Cryptosporidium – cattle, pigs, birds
  • E. coli – all animals
  • Giardia – dogs, NHP
  • Histoplasmosis – birds, bats
  • Klebsiella – fish
  • Leptospirosis – cattle, pigs, sheep, dogs, rodents
  • Listeria – cattle, sheep, birds
  • LCMV (Lymphocytic Choriomeningitis Virus) – rodents, NHP
    • Causes encephalitis
  • Mycobacterium– cattle (tuberculosis), NHP, rodents, birds
  • Pasteurella – rabbits, birds
  • Plasmodium, Entamoeba (dysentery) – NHP
  • Poxviruses – NHP, cattle, pigs, horses, birds
  • Prions (Mad Cow Disease) – sheep, goats, cattle
  • Rabies – dogs, cats, wild mammals
    • Once symptoms begin it is too late to treat
  • Ringworm and other fungal infections – cattle, dogs, cats
  • Roundworms – pigs, dogs, cats
  • Salmonella – rodents, NHP, cattle, birds, pigs, reptiles, amphibians, fish, dogs, cats
  • Shigella – NHP
  • SIV, Spuma, Foamy, Ebola, Marburg viruses – NHP
  • Staphylococcus and Streptococcus – all animals
  • Streptobacillus and Spirillium (Rat Bite Fever) – rodents
  • Tetanus – NHP, all animals may be carriers
  • Toxoplasma – cats
  • Tularemia – wild mammals

Controlling Hazards

Other Hazards

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  • Bites and Scratches
  • Scrapes or cuts from contaminated equipment
  • Accidental Needle Sticks – Animals should be effectively restrained for injections, or anesthetized if high risk agent is being used
  • Use of infectious agents – Increased risk of transmission in an animal model
  • Use of hazardous drugs, carcinogens, chemotherapeutics, controlled substances
  • Use of radiation
  • Use of recombinant DNA – Potential for off-target event with unknown effects
  • Use of human-derived materials – i.e. primary tumor cells
  • Aerosols – Can be produced when administering agents/drugs/chemicals
  • Animal Escapes – Have an SOP in place in case of an escape

Safe Sharps

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  • Avoid using sharps if possible or use safe sharps devices such as: retractable or self-sheathing needles, one-piece disposable scalpels.
  • Substitute glass with plastic, or mylar-wrapped glass.
  • Never pick up broken glass by hand, use forceps/tongs/dustpan.
  • Never bend, break, or re-cap needles – if needles must be re-capped, use a needle-recapping tool.
  • Do not remove needles or blades by hand- use forceps or a blade removal device. Use forceps or tongs when cleaning reusable sharps. 
  • Always dispose of used needles directly into sharps container
  • Do not overfill sharps container – dispose of container when ¾ full.
  • Sharps container must be puncture-proof, leak-proof, properly labeled, and sealable.
  • Refer to the Sharps Safety page for more information.

 

Engineering Controls

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When biohazards are used, work should be conducted in a biosafety cabinet for any procedure that may generate aerosols, such as:

  • Cage changes
  • Injections
  • Gavage
  • Intranasal or oropharyngeal inoculations
  • Necropsies and dissections
  • Handling of infected animals

Fume hoods should be used for any volatile chemicals
Rodent restraint devices, such as cones or tubes, may be used to help protect the user from bites or scratches while giving an injection. Anesthesia should also be considered if mechanical restraint is not possible. These methods must be approved on your IACUC protocol.

 

Good Work Practices and Hygiene

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  • Change your gloves if they become contaminated, or as required
  • Wash your hands after performing work, after you remove gloves, and just before you leave
  • Do not use cell phones, ipods, or other personal items – if they become contaminated you can carry dangerous pathogens or chemicals home with you
  • No eating, drinking, chewing gum, or applying makeup
  • Do not touch your face or hair
  • Use appropriate secondary containers and carts for transporting animals or samples

PPE

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Proper Disinfection

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  • Disinfect surfaces before and after work, use the appropriate agent and allow sufficient contact time.
  • Visit the Decontamination and Disinfection page for more information.

Minimize Aerosols

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  • Perform procedures carefully
  • Use engineering controls such as biosafety cabinets and filter top cages

Minimize Risk of Bites and Scratches

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  • Mechanical restraint devices (i.e. tube restraint for mouse procedures)
  • Use anesthesia if applicable (and if approved by the IACUC)

Enroll in the Occupational Health Program

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Occupational Health Program

Exposures and Emergency Response

Emergency Procedures

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Know how to handle an exposure or emergency before it happens. Being prepared will allow you to act quickly and efficiently in an emergency situation.

  • Flush the affected area
    • Dermal exposures - wash thoroughly with soap and water for 15 minutes
    • Eye exposures - flush at eyewash for 15 minutes
    • Encourage bleeding for small wounds. Apply pressure to large wounds.
  • Move to fresh air for inhalation exposures.
  • Know where the emergency equipment is stored, and know how to use it (first aid kit, fire extinguisher, spill kit, etc.)
  • Seek medical attention- bring the pathogen safety data sheet or SDS with you
  • Notify your supervisor as soon as possible
  • Submit a report to Risk Management