Help Registering Laboratory Strains of E. coli
All microbes used in a laboratory setting at MUSC must be registered with the IBC. The registration requirement includes common laboratory strains of Risk Group 1 E. coli, which can be worked with safely at Biosafety Level 1. This page is designed to help faculty requiring assistance registering E. coli in the microbes section (section 5) of the IBC forms.
A. Agent Identification
1. Agent/microbe: Escherichia coli (BSL1)
2. Strains: K12, TOP10, DH5-alpha, BL21
B. Agent Hazard
1. Is this a Select Agent? no
2. The agent is infectious to: mammals including humans
3. Common means / routes of transmission: injection, ingestion, autoinoculation
4. Does the agent produce a toxin? Not besides the LPS/endotoxin normally found in the E. coli cell wall.
5. What is the anticipated virulence? Risk Group 1 laboratory strains of E. coli have a low, but not non-existent, virulence. Although organisms classified as Risk Group 1 are not known to consistently cause disease in healthy adults, the following individuals may be at increased risk of infection: young, elderly, immunocompromised individuals or individuals taking large quantities of antacids or stomach acid reducing medications. Inflammation may occur if autoinoculation results from injection or contamination of broken skin.
Has the organism exhibited antibiotic resistance? These strains of E. coli are generally not resistant to antibiotics but cloning experiments generally confer antibiotic resistance to facilitate selection.
C. Experimental Procedures
Procedures will varry between labs. However key peices of information to disclose include:
- Maximum anticipated culture volume
- Method of disinfecting solid waste (plates, flasks, etc. ), liquid waste (decanted culture supernatant) and work surfaces
- Method of lysing the bacteria
Will experiments result in acquisition of new characteristics such as enhanced virulence, infectivity, drug resistance or change in host range? Three general types of answers are provided below to help investigators describe their experiments. Typical cloning experiments will fall under "b".
a. No, neither E coli nor the plasmids nor the genes used in these experiments contain resistance genes or will enhance virulence or infectivity or alter host range.
b. Yes, the plasmids confer antibiotic resistance to the following antibiotics: _____________________. Plasmids or encoded genes will not affect virulence, infectivity, or host range of E coli.
Example: Bacteria will be transformed with bacterial plasmids carrying the desired insert gene and antibiotic resistance to antibiotics commonly used in laboratories for selection (kanamycin, streptomycin, chloramphenicol, etc.), but will not interfere with potential clinical treatment. Neither plasmids nor their insert genes will affect virulence, infectivity or host range.
c. Yes, plasmids or encoded genes will affect virulence, infectivity, or host range of E coli. Please specify which is affected and how.
D. Safety Issues
1. What protective equipment will be used to minimize exposure of laboratory personnel who will be working with the agent? lab coats and gloves
2. What will be used for handwashing? Soap or alcohol gel
3. Following groups of individuals at increased risk if exposed to the agent: The following individuals may be at increased risk of infection: young, elderly, immunocompromised individuals or individuals taking large quantities of antacids or stomach acid reducing medications.
4. Is a vaccine available for the agent? No
5/6. Is collection of serum samples for banking advisable? No
7. Is there need for medical monitoring or surveillance? No
8. Please state the signs and symptoms of infection. Ingestion of E. coli may cause diarrhea with or without abdominal cramps. Skin infections may be inflammed. Conjunctivitis may occur after inoculation of the eye.
9. Please describe post-exposure therapy to be used in the event of accidental exposure. Rinse contaminated areas with soap and water. Mucous membranes should be flushed with water. For accidental ingestion rinse mouth with water, but do not swallow. Immediately notify the PI or lab manager as well as Daniel Eisenman, Biosafety Officer (792-4304, email@example.com, or pager 12883). Additional treatment, if necessary, depends on the location of the infection. Seek immediate medical care if necessary. Report incident to Employee Health Services by submitting an ACORD form.