Escherichia coli (E. coli)

Characteristics
Morphology	Escherichia coli (E. coli) are in the family Enterobacteriaceae and are Gram negative, rod shaped, non-spore forming, motile with peritrichous flagella or nonmotile, and are capable of aerobic or anaerobic growth. Many different strains exist and range from being harmless to highly pathogenic. Some non-pathogenic strains include E. coli K12, E. coli 25922, E. coli Nissle, E. coli Castellani and Chalmers ATCC 700728. Pathogenic E. coli can cause gastrointestinal infections, urinary tract infections, respiratory illness, and pneumonia. Some common strains are Shiga toxin-producing E. coli (STEC, or serotype O157:H7), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli. E. coli strains are routinely used as hosts for cloning experiments and other genetic manipulations in the laboratory.
Growth Conditions	Cultures can be placed in various growth media and incubated at 37°C (ex. LB broth or agarose plates)

Characteristics

Morphology

Escherichia coli (E. coli) are in the family Enterobacteriaceae and are Gram negative, rod shaped, non-spore forming, motile with peritrichous flagella or nonmotile, and are capable of aerobic or anaerobic growth. Many different strains exist and range from being harmless to highly pathogenic.

Some non-pathogenic strains include E. coli K12, E. coli 25922, E. coli Nissle, E. coli Castellani and Chalmers ATCC 700728. Pathogenic E. coli can cause gastrointestinal infections, urinary tract infections, respiratory illness, and pneumonia. Some common strains are Shiga toxin-producing E. coli (STEC, or serotype O157:H7), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli. E. coli strains are routinely used as hosts for cloning experiments and other genetic manipulations in the laboratory.

Growth
Conditions

Cultures can be placed in various growth media and incubated at 37°C (ex. LB broth or agarose plates)

Health Hazards
Host Range	Humans, but many strains can infect animals (both wild and domesticated)
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	None readily available
Vaccines	None readily available
Treatment	Administer appropriate antibiotic treatment in severe case. Non-specific supportive therapy, including hydration and bismuth subsalicylate compounds (e.g., Pepto-Bismol©) can help reduce recovery time and the number of bowel movements respectively.
Surveillance	Monitor for symptoms. Stool culture, immunoassays, and nucleic acid-based assays (PCR and DNA hybridization methods) are used for the detection and strain identification of E. coli 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-1/ABSL-1	Non-Pathogenic strains are associated with Risk Group 1. BSL-1 level of containment that involved standard, microbiological best practices and procedures with no special primary or secondary barriers, other than a door, a sink for handwashing, and non-porous work surfaces that are cleanable and easy to decontaminate is sufficient for these type of strains.
BSL-2/ABSL-2	Risk Group 2 classification is applied to all pathogenic strains. BSL-2 practices, containment equipment, and facilities are recommended for activities using clinical materials and diagnostic quantities of infectious cultures. It is recommended that special emphasis be placed on personal protective equipment, handwashing, manipulation of faucet handles, and decontamination of work surfaces to decrease the risk of exposure. ABSL-2 facilities and practices are recommended for activities with experimentally infected animals.

Containment

BSL-1/ABSL-1

Non-Pathogenic strains are associated with Risk Group 1. BSL-1 level of containment that involved standard, microbiological best practices and procedures with no special primary or secondary barriers, other than a door, a sink for handwashing, and non-porous work surfaces that are cleanable and easy to decontaminate is sufficient for these type of strains.

BSL-2/ABSL-2

Risk Group 2 classification is applied to all pathogenic strains. BSL-2 practices, containment equipment, and facilities are recommended for activities using clinical materials and diagnostic quantities of infectious cultures. It is recommended that special emphasis be placed on personal protective equipment, handwashing, manipulation of faucet handles, and decontamination of work surfaces to decrease the risk of exposure. ABSL-2 facilities and practices are recommended for activities with experimentally infected animals.

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. E. coli is not commonly transmitted via aerosols, but caution should 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 the bacteria (bleach, ethanol, glutaraldehyde), 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.

E. coli is not commonly transmitted via aerosols, but caution should 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 the bacteria (bleach, ethanol, glutaraldehyde), 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	For an area not protected by skin, wash with soap and water for 15 minutes (open wounds, sores, etc.)
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 10% bleach, 2% glutaraldehyde, 70% ethanol
Inactivation	Ozone can inactivate E. coli. E. coli are also sensitive to heat treatment, especially at temperatures of 70°C or higher
Survival Outside Host	E. coli has the ability to survive from hours to months on inanimate surfaces.

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 E. coli. 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. Wash hands with soap and water after removing gloves.

References
CDC (https://www.cdc.gov/ecoli/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…) ABSA (https://my.absa.org/riskgroups)