Hepatitis B virus (HBV)

Characteristics
Morphology	HBV is a member of the Hepadnaviridae family, has a circular DNA genome that is partially double stranded and partly single stranded, and is 42 nm in diameter. HBV is comprised of a number of clinically important viral proteins, including the envelope protein, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), and a soluble nucleocapsid protein, the hepatitis B e antigen (HBeAg). Eight genotypes of HBV have been identified (A to H)
Growth Conditions	PLC/PRF/5 human hepatoma cell line, HepG2, and primary human hepatocytes are commonly used for HBV growth

Characteristics

Morphology

HBV is a member of the Hepadnaviridae family, has a circular DNA genome that is partially double stranded and partly single stranded, and is 42 nm in diameter. HBV is comprised of a number of clinically important viral proteins, including the envelope protein, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), and a soluble nucleocapsid protein, the hepatitis B e antigen (HBeAg). Eight genotypes of HBV have been identified (A to H) 

Growth
Conditions

PLC/PRF/5 human hepatoma cell line, HepG2, and primary human hepatocytes are commonly used for HBV growth

Health Hazards
Host Range	Humans are the only known natural host. Chimpanzees are susceptible as an experimental animal
Modes of Transmission	HBV is transmitted through activities that involve percutaneous or mucosal contact with infectious blood or body fluids including: sharing needles, syringes, or other drug-injection equipment, sexual contact, mother to baby at birth.
Signs and Symptoms	Two major forms, asymptomatic infection and symptomatic hepatitis. Symptoms of acute infection include fever, fatigue, anorexia, abdominal discomfort, nausea and vomiting, joint pain, jaundice, clay-colored stool. Case definitions include Acute: mostly asymptomatic, or with mild to severe hepatitis, seroconversion to positive HBsAg Chronic: persistence of HBsAg for more than 6 months, increased risk for developing chronic liver disease Perinatal: transmission from mother to child Most persons with acute disease recover with no lasting liver damage; acute illness is rarely fatal, while 15%–25% of chronically infected persons develop chronic liver disease, including cirrhosis, liver failure, or liver cancer.
Infectious Dose	Unknown
Incubation Period	Usually 45-160 days; average of 90 days. HBsAg can appear in as little as 2 weeks, or as long as 6–9 months after exposure.

Medical Precautions/Treatment
Prophylaxis	Hepatitis B immunoglobulin (HBIG), can be even given within 48 hours of an exposure
Vaccines	Effective HBV vaccines are available
Treatment	Supportive treatment is an option for acute HBV infection in patients who spontaneously clear the infection. Antiviral therapy is available for severe acute and chronic infections
Surveillance	Monitor for symptoms. Three different serologic tests are needed (hepatitis B surface antigen [HBsAg], hepatitis B surface antibody [anti-HBs], and total hepatitis B core antibody [anti-HBc]) to diagnose infection
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
BSL-2/BSL-2+	Risk Group 2 classification is applied to HBV. Containment Level 2 facilities, equipment, and operational practices for work involving infectious or potentially infectious material. All procedures that may produce aerosols, or involve high concentrations or large volumes should be conducted in a biological safety cabinet (BSC). BSL-2 facilities with additional primary containment and personnel precautions, such as those described for BSL-3, may be indicated for activities with potential for droplet or aerosol production and for activities involving production quantities or concentrations of infectious materials.
ABSL-2	ABSL-2 practices, containment equipment, and facilities are recommended for activities utilizing naturally or experimentally infected chimpanzees or other non-human primates (NHPs).

Containment

BSL-2/BSL-2+

Risk Group 2 classification is applied to HBV. Containment Level 2 facilities, equipment, and operational practices for work involving infectious or potentially infectious material. All procedures that may produce aerosols, or involve high concentrations or large volumes should be conducted in a biological safety cabinet (BSC). BSL-2 facilities with additional primary containment and personnel precautions, such as those described for BSL-3, may be indicated for activities with potential for droplet or aerosol production and for activities involving production quantities or concentrations of infectious materials.

ABSL-2

ABSL-2 practices, containment equipment, and facilities are recommended for activities utilizing naturally or experimentally infected chimpanzees or other non-human primates (NHPs).

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. While HBV exposure via aerosol transmission is rare, extra caution should still be taken during a spill outside of a BSC. 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 enveloped viruses (alcohols, bleach, and quaternary ammonium compounds), working from the perimeter towards the center. Allow 30 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.

While HBV exposure via aerosol transmission is rare, extra caution should still be taken during a spill outside of a BSC. 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 enveloped viruses (alcohols, bleach, and quaternary ammonium compounds), working from the perimeter towards the center. Allow 30 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	Susceptible to 70% Ethanol, 10% bleach, and Quaternary ammonium compounds
Inactivation	Moist heat at 98°C for 1 minute will partially inactivate HBV in a 1:10 serum dilution. Incubation at 60°C for 10 hours (pasteurization) will also inactivate HBV
Survival Outside Host	Survives in dried blood for long periods (weeks), stable on environmental surfaces for a least 7 days at room temperature.

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 HBV. Additional PPE may be required depending on lab specific SOPs.
Additional Precautions	Additional protection may be worn over laboratory clothing when infectious materials are directly handled, such as solid-front gowns with tight fitting wrists, gloves, and respiratory protection.

References
CDC (https://www.cdc.gov/hepatitis/hbv/) BMBL 6th Edition (https://www.cdc.gov/labs/BMBL.html) Cell Culture Models for the Investigation of Hepatitis B (https://www.mdpi.com/1999-4915/8/9/261) Canadian PHAC (https://www.canada.ca/en/public-health/services/laboratory-biosafety-bi…)