Adenovirus and Adenoviral Vectors

Characteristics
Morphology	A member of the Retroviridae family, genus Lentivirus. Human Immunodeficiency Virus (HIV) is an icosahedral, enveloped virus, of approximately 100 to 110 nm in diameter, and has a single- stranded, linear, positive-sense RNA genome. HIV has two recognized strains, HIV-1 and HIV-2
Growth Conditions	Primarily T-cell based cell lines (ex. SupT1, Jurkat, and CEM)

Health Hazards
Host Range	Humans and animals are susceptible hosts to wildtype AdV. The host range of engineered AdV vectors will depend on the target gene and promoters engineered, as a result a wide variety of dividing and non-dividing cells can be transformed.
Modes of Transmission	Respiratory and fecal-oral routes. AdV can also spread through contaminated fomites, ingestion, and inhalation of aerosolized droplets.
Signs and Symptoms	AdV infection causes a mild respiratory tract infection (resembling a common cold or flu) which is self-limiting and generally asymptomatic. Infections may also affect the gastrointestinal tract, and eyes (conjunctivitis). Symptoms may include fever, nasal congestion, rhinitis, and pharyngitis. AdV and AdV vectors can induce varied immunological responses in the host, depending on the serotype.
Infectious Dose	Inhalation of as few as 5 Adenovirus particles can cause disease in susceptible individuals.
Incubation Period	Usually 2 to 14 days

Medical Precautions/Treatment
Prophylaxis	Maintain proper hygiene and wash hands; disinfect any surfaces that might have been exposed
Vaccines	None publically available
Treatment	Most adenovirus infections are mild and may require only care to help relieve symptoms
Surveillance	Monitor for symptoms. Laboratory confirmation is done via immunofluorescence, enzyme-linked immunoassay, or PCR.
GWU Requirements	Report all incidents to the Office of Risk Management as well as the Office of Research Safety (ORS) IMMEDIATELY FOLLOWING THE OCCURRENCE.

Containment
BSL2/ABSL2	Risk Group 2 classification is applied to the species as a whole, but may not apply to every serotype. Containment Level 2 facilities, equipment, and operational practices. No open-bench work should be performed with AdV wild type or vectors. All work should be performed inside a Biosafety Cabinet. Centrifuge rotors must have a lid, samples should be loaded/unloaded inside the BSC and the centrifuge should be decontaminated with appropriate disinfectant after use. If the vector is replication incompetent, animals infected with AdV vectors will remain at ABSL-2 for 72h, then moved to ABSL1. If the vector is replication competent, animals will be housed at ABSL2 for the length of the experiment.

Containment

BSL2/ABSL2

Risk Group 2 classification is applied to the species as a whole, but may not apply to every serotype. Containment Level 2 facilities, equipment, and operational practices. No open-bench work should be performed with AdV wild type or vectors. All work should be performed inside a Biosafety Cabinet. Centrifuge rotors must have a lid, samples should be loaded/unloaded inside the BSC and the centrifuge should be decontaminated with appropriate disinfectant after use. If the vector is replication incompetent, animals infected with AdV vectors will remain at ABSL-2 for 72h, then moved to ABSL1. If the vector is replication competent, animals will be housed at ABSL2 for the length of the experiment.

Spill Procedures
Small Spills (<1 liter)	If the spill occurred inside a biological safety cabinet, close the sash and allow the cabinet to operate for 15 minutes before continuing with the spill cleanup. Adenovirus and Adenoviral vectors can be transmitted via aerosols. If the spill occurred outside of a biological safety cabinet, leave the room immediately and allow the aerosols to dissipate for 15 minutes. Notify others working in the lab. Don appropriate PPE. Cover area of the spill with paper towels or any absorbent material and apply an EPA registered disinfectant effective against non-enveloped viruses (such as peroxide, bleach, and Cavicide), working from the perimeter towards the center. Allow 15-20 minutes of contact time before disposal and cleanup of spill materials.
Large Spills	Alert lab personnel in the laboratory to the spill and keep people out of the area to prevent spread of the contamination. Check if you have been contaminated or if any of your PPE has been breached. If so follow exposure procedures. Remove any contaminated clothing and place it the biohazard waste. Wash your hands and post a sign on the door. Notify your supervisor of the incident and call ORS (4-8258) for assistance. If the situation involves an imminently life-threatening injury or has catastrophic potential, call 911.

Spill Procedures

Small Spills (<1 liter)

If the spill occurred inside a biological safety cabinet, close the sash and allow the cabinet to operate for 15 minutes before continuing with the spill cleanup.

Adenovirus and Adenoviral vectors can be transmitted via aerosols. If the spill occurred outside of a biological safety cabinet, leave the room immediately and allow the aerosols to dissipate for 15 minutes. Notify others working in the lab. Don appropriate PPE. Cover area of the spill with paper towels or any absorbent material and apply an EPA registered disinfectant effective against non-enveloped viruses (such as peroxide, bleach, and Cavicide), working from the perimeter towards the center. Allow 15-20 minutes of contact time before disposal and cleanup of spill materials.

Large Spills

Alert lab personnel in the laboratory to the spill and keep people out of the area to prevent spread of the contamination. Check if you have been contaminated or if any of your PPE has been breached. If so follow exposure procedures. Remove any contaminated clothing and place it the biohazard waste. Wash your hands and post a sign on the door. Notify your supervisor of the incident and call ORS (4-8258) for assistance. If the situation involves an imminently life-threatening injury or has catastrophic potential, call 911.

Exposure Procedures
Mucus Membrane	Flush eyes, mouth or nose for 15 minutes at eyewash station.
Other Exposure	Wash area with soap and water for 15 minutes.
Reporting	Report ALL injuries to the PI immediately and reported to the Office of Risk Management at [email protected] IMMEDIATELY FOLLOWING THE OCCURRENCE. Exposures that involve a bloodborne-pathogen or recombinant DNA also need to be reported to the Office of Research Safety at [email protected]. If the injury requires immediate medical attention, call GWPD at 202-994-6111 or call 911.
Medical Monitoring	Seek immediate medical evaluation, treatment, and post exposure follow-up at the Employee Health Office at GWU Hospital (900 23rd St., NW, Suite G-1090, Phone: 202-715-4275). Students should go to the Students Health Office at Marvin Center. After hours treatment can be received at the GWU hospital emergency room.

Stability
Disinfection	10% fresh bleach solution, a minimum of 10 min contact time is required. Alcohol alone is NOT an effective disinfectant against AdV.
Inactivation	AdV can be inactivated by heat (56°C for 30 min, 60°C for 2 min) and by autoclaving.
Survival Outside Host	AdV is very stable in the environment and persist for 7 days to 3 months on dry inanimate surfaces at room temperature. Higher temperatures decrease this time, but most serotypes are stable up to 7 days even at 36°C. Most can also survive for weeks in tap water, sewage effluent and sea water.

Personal Protective Equipment (PPE)
Minimum PPE Requirements	At minimum, personnel are required to don gloves, closed toed shoes, lab coat, and appropriate face and eye protection prior to working with Adenovirus. Additional PPE may be required depending on lab specific SOPs.
Additional Precautions	Use respiratory protection if work will be performed outside the biosafety cabinet. Additional precautions should be considered with work involving animals or large scale activities.

References
CDC (https://www.cdc.gov/adenovirus/) Canadian PHAC (https://www.canada.ca/en/public-health/services/laboratory-biosafety-bi…)