Rabies

Characteristics
Morphology	Rabies is an acute progressive encephalitis caused by the rabies virus (RABV), a bullet-shaped, enveloped virus of approximately 75 nm in diameter by 180 nm in length, and has a single-stranded, negative-sense RNA genome. It is part of the family Rhabdoviridae, genus Lyssavirus, and has 7 members, of which only serotype 1 commonly infects humans, while the other 6 are rare causes of human disease
Growth Conditions	BHK-21 and Vero cell lines

Characteristics

Morphology

Rabies is an acute progressive encephalitis caused by the rabies virus (RABV), a bullet-shaped, enveloped virus of approximately 75 nm in diameter by 180 nm in length, and has a single-stranded, negative-sense RNA genome. It is part of the family Rhabdoviridae, genus Lyssavirus, and has 7 members, of which only serotype 1 commonly infects humans, while the other 6 are rare causes of human disease

Growth
Conditions

BHK-21 and Vero cell lines

Health Hazards
Host Range	Humans, and many mammals, most commonly wild and domestic canids
Modes of Transmission	Rabies is most commonly transmitted to humans via the bite of a rabies-infected animal. Bites to the head, neck, and arms are the most likely to lead to transmission. It is also possible, but rare, for people to get rabies from non-bite exposures, which can include scratches, abrasions, or open wounds that are exposed to saliva or other potentially infectious material from a rabid animal. Inhalation of aerosolized rabies virus is one potential non-bite route of exposure, but except for laboratory workers, most people won’t encounter an aerosol of rabies virus.
Signs and Symptoms	The first symptoms of rabies may be similar to the flu, including weakness or discomfort, fever, or headache. There also may be discomfort, prickling, or an itching sensation at the site of the bite. These symptoms may last for days. Symptoms then progress to cerebral dysfunction, anxiety, confusion, and agitation. As the disease progresses, the person may experience delirium, abnormal behavior, hallucinations, hydrophobia (fear of water), and insomnia. The acute period of disease typically ends after 2 to 10 days. Once clinical signs of rabies appear, the disease is nearly always fatal, and treatment is typically supportive.
Infectious Dose	Unknown
Incubation Period	Varies from days to more than 7 years, with 75% of patients becoming ill within 90 days of exposure.

Medical Precautions/Treatment
Prophylaxis	Post-exposure rabies prophylaxis with rabies vaccine together with the administration of rabies immunoglobulin (RIG) is highly effective
Vaccines	Imovax Rabies, a human diploid cell vaccine (HDCV), or RabAvert, a purified chick embryo cell vaccine (PCECV) are available.
Treatment	Treatment starts with good wound care, which can reduce the risk of rabies by up to 90%. Wash the wound with a soap solution, followed by 70% ethanol or an iodine containing solution. Post-exposure rabies vaccination can then be administered to reduce serious symptoms.
Surveillance	Monitor for symptoms. Rabies is diagnosed using the direct fluorescent antibody (DFA) test, which looks for the presence of rabies virus antigens in brain tissue. Saliva can be tested by virus isolation or reverse transcription followed by polymerase chain reaction (RT-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 3 classification is applied to RABV. BSL-2 and/or ABSL-2 practices, containment equipment, and facilities are recommended for all activities utilizing known or potentially infectious materials or animals. Pre-exposure rabies vaccination is recommended for all individuals prior to working with lyssaviruses or infected animals or engaging in diagnostic, production, or research activities with these viruses.
BSL-3/ABSL-3	Primary containment and personnel precautions, such as those described for BSL-3 and/or ABSL-3, are indicated for activities with a high potential for droplet or aerosol production, and for activities involving large production quantities or high concentrations of infectious materials/animals. Certain Lyssavirus species should utilize BSL-3 containment procedures (Lagos bat lyssavirus, Mokola lyssavirus, Shimoni bat lyssavirus, and West Caucasian bat lyssavirus).

Containment

BSL2/ABSL2

Risk Group 3 classification is applied to RABV. BSL-2 and/or ABSL-2 practices, containment equipment, and facilities are recommended for all activities utilizing known or potentially infectious materials or animals. Pre-exposure rabies vaccination is recommended for all individuals prior to working with lyssaviruses or infected animals or engaging in diagnostic, production, or research activities with these viruses.

BSL-3/ABSL-3

Primary containment and personnel precautions, such as those described for BSL-3 and/or ABSL-3, are indicated for activities with a high potential for droplet or aerosol production, and for activities involving large production quantities or high concentrations of infectious materials/animals. Certain Lyssavirus species should utilize BSL-3 containment procedures (Lagos bat lyssavirus, Mokola lyssavirus, Shimoni bat lyssavirus, and West Caucasian bat lyssavirus).

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. RABV exposure via aerosol transmission is a possibility, so 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 (bleach, povidone-iodine, and chlorhexidine), 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.

RABV exposure via aerosol transmission is a possibility, so 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 (bleach, povidone-iodine, and chlorhexidine), 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	Rabies virus is inactivated by exposure to 70% ethanol, phenol, formalin, ether, trypsin, β-propiolactone, and some other detergents
Inactivation	Rabies virus does not tolerate pH below 3 or above 11, autoclaving, and is inactivated by ultraviolet light
Survival Outside Host	This virus does not survive well outside its host (in dried blood and secretions) as it is susceptible to sunlight and desiccation.

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 RABV. Additional PPE may be required depending on lab specific SOPs. Higher containment levels will require additional precautions.
Additional Precautions	Additional protection may be worn over laboratory clothing when infectious materials are directly handled and when working at higher containment levels, such as solid-front gowns with tight fitting wrists, gloves, and respiratory protection. Eye protection must be used where there is a known or potential risk of exposure to splashes

References
CDC (https://www.cdc.gov/rabies/index.html) BMBL 6th Edition (https://www.cdc.gov/labs/BMBL.html) Canadian PHAC (https://www.canada.ca/en/public-health/services/laboratory-biosafety-bi…) ATCC (https://www.atcc.org/products/vr-959) ABSA (https://my.absa.org/riskgroups) Epidemiology of Rabies and Current US Vaccine Guidelines (https://pubmed.ncbi.nlm.nih.gov/32752569/)