UK Policies Biological Safety

Originally approved by the UK Institutional Biosafety Committee (IBC) on May 12, 2010, the following requirements for tissue culture rooms must be met:

1. Air shall flow from the hallway to the inner lab (negative to the hallway). All room air shall be exhausted through ducts to the outside of the building and not recirculated within the building. Room air which is exhausted to a common plenum is NOT acceptable.
2. The number of BSCs, the amount of space within the BSC and amount of room space provided is required to accommodate all of the tissue culture and viral vector work. This is for the protection of the research materials and for the protection of the researchers and facility.
3. The BSCs may be recirculating models (Class II, A2) or thimble ducted or hard ducted.
   1. BSCs shall be installed such that:
      1. Fluctuations of the room air supply and exhaust do not interfere with proper operations.
      2. Manufacturers’ guidelines are followed.
      3. They can be certified follow the National Sanitary Foundation (NSF) criteria.
   2. BSCs shall be certified on an annual basis by a vendor who meets the requirements of the Biological Safety Department, follows NSF criteria, and is on contract with UK.
      1. Coordination of this certification is through the Biological Safety Department.
      2. Payment for this service is the responsibility of the Principal Investigator or Department
4. Rooms housing any BSC shall be configured to allow storage of supplies and equipment used with the biohazardous materials. Typical equipment in these rooms includes: incubators, centrifuges, microscopes, CO2 tanks, vacuum source, refrigerators.
5. A hand washing sink with eyewash shall be present in the tissue culture room facility. The eyewash may be “drench hose” if approved by UK Occupational Health and Safety Department. An exemption from the eyewash requirement may be granted by the Biological Safety Officer if the risk assessment of the proposed research warrants it.
6. There must be restricted entry to the outer laboratory, which is locked when no one is present.
7. All surfaces must be easily cleaned and decontaminated. Room casework shall be easily cleanable, and finishes should be compatible with materials used for cleaning and disinfection. Chairs must be covered with a non-porous material. Rugs or carpets are not permitted.
8. Vacuum lines shall be protected with High Efficiency Particulate Air (HEPA) filters or their equivalent. This applies to central vacuum systems and to individual vacuum pumps.
9. Open flames SHALL NOT be used in BSCs. Therefore, gas lines SHALL NOT be connected to BSCs.
10. A functioning and validated autoclave enrolled in the UK Autoclave Verification program shall be available within a reasonable distance of the facilities creating the biohazardous waste.
    1. IBC approved procedures for transport shall be followed when unprocessed waste carried through public hallways and elevators.
11. Tissue culture rooms which will contain research deemed by the IBC to be BSL2+ (enhanced) shall be in an inner lab, with two doors between the BSC and the hallway.
12. Appropriate signage shall be displayed on the door of the main laboratory and of the tissue culture room.

• Vaccination IS NOT RECOMMENDED for researchers working with highly attenuated strains of vaccinia virus (MVA, NYVAC, TROVAC, ALVA), with limited exceptions.
• Vaccination IS REQUIRED for researchers working directly with non-highly attenuated strains of vaccinia virus (WR, NYCBOH, Copenhagen, Temple of Heaven, Lister), or Cowpox or Monkeypox OR animals infected with those strains.
• An unvaccinated worker shall not work directly with non-highly attenuated vaccinia virus OR animals infected with those strains.
• Other factors, such as researcher parameters, also influence vaccination recommendations.
• Documented confidential medical counseling by University Health Service IS REQUIRED for anyone who will be working in a laboratory where vaccinia virus (highly attenuated or non-highly attenuated) is manipulated.

I. Introduction: 

Recombinant vaccinia and other pox viruses are useful microbiological research tools for expression of exogenous proteins in a variety of cultured cell types. However, their use is not without risk to laboratory personnel. As stated in the Centers for Disease Control (CDC) and National Institutes of Health's Biosafety in Microbiological and Biomedical Laboratories 5th edition (http://www.cdc.gov/od/ohs/biosfty/bmbl5toc.htm, Section VIII-E) and the Vaccinia (Smallpox) Vaccine Recommendations of the Advisory Committee on Immunization Practices (ACIP), 2001 (http://www.cdc.gov/mmwr/preview/mmwrhtml/rr5010al.htm):

• Naturally or experimentally infected laboratory animals are a potential source of infection to exposed unvaccinated laboratory personnel.
• Genetically engineered recombinant vaccinia viruses pose an additional potential risk to laboratory personnel, through direct contact or contact with clinical materials from infected volunteers or animals.
• The agents may be present in lesion fluids or crusts, respiratory secretions, or tissues of infected hosts.
• Ingestion, parenteral inoculation, and droplet or aerosol exposure of mucous membranes or broken skin with infectious fluids or tissues, are the primary hazards to laboratory and animal care personnel.
• All manipulations of vaccinia strains BSL-2 and above should be conducted in a biosafety cabinet. When work must be performed outside of a biosafety cabinet (e.g. animal surgery, microscopy), the following personal protective equipment must be used: disposable gloves, disposable lab coat or gown, eye and mucous membrane protection (goggles which are ANSI certified and surgical mask or face shield). Serious ocular infections can occur even in vaccinated individuals.

Multiple strains of vaccinia virus exist with varying levels of virulence for humans and animals. Depending on the strain used, vaccinia virus presents varying levels of health risk to laboratory personnel. Strains that are highly attenuated are typically unable to replicate or replicate poorly in human cells. On the other hand, non-highly attenuated strains of vaccinia have the ability to replicate in human cells and thus pose a risk to the public health. Risk include localized skin infections and more severe, disseminated reactions to which immunocompromised individuals may be more susceptible.

Routes of transmission for occupationally acquired infections of vaccinia:

1. needlestick injury
2. contamination of non-intact skin, mucous membranes, eyes
3. unknown exposure, possibly contamination of intact skin which was not immediately washed
4. Potential routes: ingestion, inhalation, exposure to fomites (virus is stable at room temperature)

II. Vaccination Recommendations and Requirements:

The following information is based on national guidelines issued by the CDC in Vaccinia (Smallpox) Vaccine Recommendations of the Advisory Committee on Immunization Practices.

Vaccination is not recommended for those working with the following highly attenuated strains:

*Biosafety level may increase depending on the presence and characteristics of a foreign protein expressed by a recombinant vaccinia virus or other aspects of the proposed experiment.

• Laboratory personnel who work with highly attenuated strains of vaccinia virus (e.g., MVA and NYVAC) or who work with the Avipoxvirus strains ALVAC and TROVAC do not require routine vaccinia vaccination.
• The Occupational Safety Health Board of NIH no longer requires vaccinia vaccination for personnel manipulating MVA or NYVAC in laboratories using only those strains.
• The Recombinant DNA Advisory Committee of the NIH reduced the biosafety level of NYVAC, TROVAC and ALVAC to level 1 based on accumulated attenuation data and biological properties of these strains.
• Although there is no formal surveillance system in place, there have not been any reports of laboratory-acquired infection resulting from exposure to any of the above highly attenuated strains or recombinant vaccines derived from these strains in the literature or to the CDC.
• Appropriate biosafety guidelines and infection control procedures should always be observed when working with viral material even if vaccination is not indicated.

There are some limited exceptions to this recommendation when working with highly attenuated strains, depending upon other factors which may come into the evaluation whether to vaccinate or not. Such as:

• The preparation of large volumes of high titer vaccinia virus.
• Work with recombinant vaccinia viruses that might have enhanced virulence or that express harmful proteins.
• Injection of high titer vaccinia virus into animals or other work with sharps and the vaccinia virus.
• Other potentially hazardous manipulations of the virus.

Vaccination is required for laboratory workers who directly handle a) cultures or b) animals infected with:

• Non-highly attenuated Vaccinia virus strains
• Recombinant Vaccinia viruses derived from non-highly attenuated vaccinia strains
• Other orthopox viruses that can infect humans

III. Advantage and Disadvantages of Vaccination:

Post-vaccination complications are possible. The risks and benefits of the vaccine have to be weighed against the exact duties of the worker. Therefore, a mandatory counseling session is required before any worker accepts or declines the vaccinia vaccination. The Principal Investigator is required to make arrangements with the University Health Service/Employee Health Service (323-582-3228) for all laboratory workers who would work with the vaccinia virus to receive this counseling. All medical information is deemed confidential and cannot be disclosed by the UHS/EHS to supervisors, etc. without the employee's written permission.

Some of the arguments for the advantages of vaccination:

1. It confers some protection in the event of an incident. (Vaccinia immune globulin (VIG) is also available through the CDC as an Investigational New Drug (IND)).
2. It may reduce the risk of seroconversion to genetically inserted material such as protein products of inserted gene material.
3. It may reduce the risk of serious eye infections following accidental splash (this risk could also be mitigated by wearing proper personal protective equipment).

Some of the arguments for the disadvantages of vaccination:

1. Risk of side-effects, which may be greater for primary rather than secondary vaccinees and for adults rather than children.
2. It does not always offer full protection.
3. Strict personal hygiene precautions must be followed for at least two weeks after vaccination. The vaccinia virus can be spread to close contacts which is especially dangerous for certain populations: infants, children, elderly, people who are immunocompromised, people with a history of eczema or atopic dermatitis, people who are pregnant or breastfeeding, people with allergies to components of the vaccine (which my be trace amount of polymyxin B, streptomycin, tetracycline, neomycin, glycerin, phenol).

IV. Additional Considerations for Vaccination:

• Revaccination every 10 years is recommended by the CDC for people working with non-highly attenuated vaccinia strains; more frequent revaccination may be required for more virulent orthopox viruses.
• Laboratory personnel not directly handling cultures of vaccinia or animals infected with vaccinia, but working in the same lab where non-highly attenuated strains are being used should be offered medical screening for potential contraindications to vaccinia exposure.
• Other health-care workers (such as physicians and nurses) working with vaccinia virus and whose contact with these viruses is limited to contaminated materials (for example, dressings), but who adhere to appropriate infection control measures, are probably at lower risk for inadvertent infection than laboratory workers. However, because a theoretical risk of infection exists, vaccination may be considered for this group.
• A summary of published case reports of laboratory-acquired vaccinia virus infections is available in the Journal of the American Biological Safety Association, Applied Biosafety, 10(2) 2005, p.118-122 by Karen Byers.

V. University of Kentucky Policy Statement:

In interest of providing a safe workplace, to comply with federal regulations, and to protect the public health, the Institutional Biosafety Committee (IBC) has formulated a policy regarding immunization for workers in laboratories using vaccinia and other pox viruses. Recommendations and requirements for vaccination will be dependent upon the strain used and procedures in the proposed research. The IBC policy incorporates national guidelines set forth by the CDC as described above and as instituted by the CDC and NIH at their own facilities.

Based on these guidelines, laboratory personnel for whom vaccination is recommended or required must receive mandatory confidential medical counseling before beginning worth with the virus. These individuals must be counseled on the risks and benefits of the vaccine and medically screened for contraindications to vaccinia exposure or vaccination.

VI. Instructions to Principal Investigators for obtaining IBC approval to use vaccinia virus and receiving vaccination:

1. Principal Investigators (PI) must register research involving vaccinia virus work with the IBC
2. PI's who have proposed research activities involving vaccinia must contact University Health/Employee Health Service (UHS) to arrange for counseling of all laboratory workers who would be in a laboratory where the virus (highly attenuated or non-highly attenuated) will be manipulated or where animals infected with vaccinia are held or manipulated.

3. If research will involve non-highly attenuated vaccinia virus, the PI must also contact UHS to arrange for acquisition of the vaccine and vaccination of all laboratory workers who would be working directly with the virus or with animals infected with the virus, even if they think they do not want to be vaccinated.

   Following medical consultation, the individual will decide whether to receive the vaccination or not.

   a. If they decide to receive the vaccination, they will sign a vaccinia vaccination consent form and receive the vaccine (This will require follow-up visits to monitor the vaccination site.) OR,

   b. If they decide not to receive the vaccine, or if UHS determines that vaccination is contraindicated, they must sign a declination form, the PI will be contacted by UHS and alterations in the individual's lab or clinical duties and responsibilities will be made such that they will not directly handle vaccinia or animals infected with vaccinia, in order to protect the health and safety of that person, their contacts, and the public health.

4. If the individual does not take the vaccination, for whatever reason, and other duties cannot be found in the laboratory such as to prevent potential contact with non-highly attenuated vaccinia, the employee must resign from his/her position. UK Human Resources will attempt to help the individual find another comparable position at the University, however a position is not guaranteed.
5. IBC approval may occur before all personnel listed on the IBC registration form have been counseled and vaccinated. It is the Principal Investigator's responsibility to ensure that all personnel working directly with vaccinia have received counseling and vaccination. Documentation of medical counseling of all laboratory workers. whether they will work directly with the vaccinia virus or simply work in the laboratory, must also be present in the PI's laboratory biosafety manual. Copies of this documentation must be sent to the Biological Safety Officer.