A biological toxin is a toxic material or product of plants, animals, microorganisms (including, but not limited to, bacteria, viruses, fungi, or protozoa). Select Agent Toxins (SAT) or Select Toxins (ST) are biological toxins or infectious substances, or a recombinant or synthesized molecule, whatever their origin and method of production. SATs can be used to endanger human and/or agricultural animal or plant health. The Center for Disease Control and Prevention (CDC) and the Animal and Plant Health Inspection Service (APHIS) of the United States Department of Agriculture (USDA) are charged with regulating these toxins.
The Biosafety Program helps MIT PIs and other personnel to avoid obtaining quantities of select agent toxins (SAT) that will cause them to inadvertently exceed the per PI exclusion limit and become subject to and violate the federal regulations. The purchase of any SAT quantity must be done through the Biosafety Program.
For more information about Select Agent Toxins and associated regulation please visit https://www.selectagents.gov/regulations.html
- Which biological toxins are Select Agent Toxins and what are the maximum limits I can have?
- Is there a protocol for using Select Agent Toxins at MIT?
- Must I be registered to use a Biological Toxin and are there any training requirements?
- How do I purchase a Select Agent Toxin?
- What are the safety concerns?
- What records am I required to keep?
- How must I store my Toxins?
- What are my responsibilities for securing my Toxins?
- How do I dispose of my Toxins?
- Can I ship Toxins or receive them directly from a collaborator or vendor?
- Can I transfer possession, distribute, or share my Toxins?
- What if I suspect a theft, loss, or release of my Toxin?
(*) Means the aggregate amount of the toxin under the control of a principal investigator, that does not, at any time, exceed the amounts in the above table.
For a list of excluded Select Agent Toxins visit https://www.selectagents.gov/SelectAgentsandToxinsExclusions.html
Please refer to Standard Operating Procedure titled “Select Agent Toxin Control and Containment Program: Purchasing, Inventory, Shipping, Receiving and Other Procedures” (EHS-0025) for full details.
Principal Investigators must be registered to work with a biological toxin. Registration is typically done through an amendment to the PI’s Biological Research Registration. This process allows EHS Biosafety and the Committee on the Assessment of Biohazards (CAB-ESCRO) to review the proposed research and evaluate any safety and regulatory concerns. Please see our Registration webpage for more details on this process.
Prior to working with a Select Agent toxin, each researcher must complete Use of Biological Toxins Training (EHS00215C). This training is given on a quarterly basis at the EHS office. Researchers can register through the MIT Atlas Learning Center. Please see the Biosafety Specific Training page for more details on training.
- Select Agent toxins (SAT) must be ordered through MIT's EHS Office, Biosafety Program via the use of a paper requisition. Requisitions can be emailed to firstname.lastname@example.org to the attention of Program Manager.
- Paper requisition must be signed and approved by the PI and include the following information
- The company’s name, address and telephone number
- The material name, catalogue number and amount ordered
- The name and laboratory/location of the PI, and where the toxin is to be delivered
- The current inventory of this select agent toxin in the laboratory (total of existing amount before the current order)
- Cost Object and account number for funds to pay for purchase
- A copy of the last current page of the inventory log for the select toxin
- The MIT EHS Office will verify that the requisition is properly authorized and the order does not inadvertently put the laboratory or its personnel at risk of exceeding per PI toxin amount limit and of becoming subject to and violating the federal regulations.
- Final approval and verification are contingent upon receipt of all requested information. (For detailed information refer to SOP EHS 0025, section 5.1). Orders will be placed within 24 hours Monday through Thursday (except holidays). Laboratories may contact the Biosafety Program Deputy Director (DD) or SAT Program Manager if a special request is needed.
- The order will shipped to "EHS Office" at MIT Building N52-496.
- The order will be received, verified against the original order. The order will be delivered to your laboratory by BSP personnel on the same day.
- When the order is released, the PI or designee will be required to sign off. Upon receipt, laboratory personnel and BSP shall immediately verify quantity received by checking the content of the vial(s) visually, log amount of new stock material into the inventory and lock up stock materials.
Authorized and EHS trained laboratory personnel should be mentored and supervised by a more experienced researcher in the safe handling of select toxin by using a current written protocol and/or Standard Operating Procedure (SOP) detailing the technique/experiments steps/handling, containment equipment, personal protective equipment (PPE), storage, disposal, internal communication and emergency response. This protocol or SOP should be updated as needed and kept accessible to users.
If at all possible, order toxins in liquid form rather than as a powder. Powders present the greatest risk during preparation of stock solutions due to possible aerosolization and danger of inhalation during handling. If the toxin is only available in powder form, order the toxin in vials that provide safety features (see table below). Avoid ordering toxins in powder form in scaled glass ampoules, as scaled glass ampoules are the most difficult to handle safely.
Some of the most commonly used vendors sell toxins in screw cap vials. These must also be handled carefully making sure the powder remains pelleted at the bottom of the tube. This can be accomplished by quickly centrifuging the vial.
- Put on personal protective equipment (PPE):
- Gloves: Gloves shouldn’t be permeable to toxin or solvent; double gloving is preferable
- Lab coat
- Safety goggles to protect from splash and/or impact
- When handling concentrated toxin, work inside a containment device such as a chemical fume hood or Biosafety Cabinet (BSC):
- Remove any unneeded chemicals, satellite accumulation areas (SAAs), or unnecessary equipment (as reasonable)
- Line the work surface of the chemical fume hood or BSC with a plastic backed absorbent liner
- If the toxin comes in regular screw cap vials:
- Quickly centrifuge the tube to pellet the powder
- Place the vial into a rack or tube holder in the chemical fume hood or BSC
- Open the screw cap vial carefully
- If the toxin comes with a rubber septum (including screw caps):
- Do not remove the rubber septum
- Use a blunt-tipped needle or pipet tip to inject measured amount of buffer/solvent through the rubber membrane
- Cover rubber septum with parafilm and mix carefully to avoid aerosol generation
- Briefly spin in a centrifuge to bring the liquid on the inside of the cap or near the lid
- If the toxin comes in a pre-weighed amount in a glass ampoule:
- Nick the ampoule with a file or file at the narrow end of the neck
- Wrap the ampoule in disinfectant-wetted cotton or paper towel
- Hold the ampoule away from you
- Snap the ampoule to open at the nick
- For other containers:
- Inject measured amount of buffer/solvent into the vial/ampoule with a pipette tip or blunt-tipped needle
- Dissolve the powder and mix liquid contents carefully to avoid aerosol generation
- Mix to give a known concentration stock solution.
- Stock solution division
- Use a pipette tip to remove contents
- Divide the solution into multiple tubes to reduce repeated freeze-thaw cycles
- These are the stock solutions, with a known amount of toxin in a known volume
- Enter the volume and number of tubes of the stock solution in a new inventory sheet. You can get new sheets here.
- Store all stock solutions in a secured lock box.
- Waste handling:
- Treat the glass vial, ampoule or screw cap vial with an appropriate concentration of disinfectant (see your “Toxin Use SOP”) for the appropriate contact time
- Dispose of glass vial(s), ampoule(s), needles, syringes in a puncture proof sharp container
- Once full, these containers are be capped tightly/sealed and placed inside a biowaste box for disposal
- Wipe down chemical fume hood or BSC surface with an appropriate concentration of disinfectant.
- Collect all solid waste (gloves, plastic backed absorbent paper) in a plastic bag and dispose in biowaste box
- If contamination is significant (such as in the case of a spill of concentrated toxin), this waste should be segregated as chemical hazardous waste. Please contact BSP if this situation arises for further guidance and coordination on the proper disposal. Under no circumstance must any select toxin waste reach public sewage systems.
The following records should be maintained at the laboratory’s location:
- A toxin inventory sheet (click appropriate link below for templates):
- Copies of each toxin purchase order
- Records are kept by Principal Investigator for at least 3 years after the select toxin is completely used up or is destroyed
Storage of select toxins shall comply with the following criteria:
- Storage shall be secured (see section 8 of SOP EHS-0025 for more details).
- Shall be tampering proof, e.g. tubes, bottles, aliquots should be clearly labeled and numbered for rapid identification of material. Information should include, volume, date of preparation, vial # and user/prepared name.
- Tubes should be, if possible, rubber septum with screw or crimped caps for added safety and should be wrapped with parafilm and kept in a spill proof secondary container e.g. properly labeled zipper storage bag or box.
- Inventory shall be accurate and up-to-date.
- Access should only be given to properly trained personnel (for more information, see section 4 of SOP EHS-0025).
For long term storage requirements see section 10.2 of SOP EHS-0025.
Custody of select agent toxins is the responsibility of the PI once EHS-BSP delivers the select agent toxins to the approved and trained laboratory personnel. All stock solutions of select agent toxins are kept under lock and key. Diluted working solutions may be stored outside of the lock box but the amounts of diluted solutions should be minimized.
For more information consult SOP EHS-0025.
Investigators shall only dispose of toxins using EHS BSP approved methods. For detailed information please see Appendix C: Toxin inactivation and disposal of SOP EHS-0025 (Please click link to access the SOP). All generated select toxin liquid waste shall be treated as hazardous chemical waste and labeled with a “red tag” as “inactivated toxin” with appropriate required information before offering it for pick up by EHS.
Solid and sharp wastes shall be assessed to determined level of toxin residue before disposing in the proper waste stream (e.g. biowaste box, puncture proof sharp container, or as chemical hazardous waste where appropriate). If necessary, researchers are encouraged to contact EHS-BSP for assistance with this assessment.
MIT Laboratories are not certified or authorized to ship via carrier (e.g. FedEx), courier, hand-carried or public transportation without assistance from EHS certified shippers; or receive select toxins under any circumstances without EHS-BSP Office involvement.
PI's shall not allow purchase of select toxins for stock piling, distribution, transfer or sharing. PIs are expected to acquire select toxins ONLY through EHS-BSP, use all of the select toxins under their custody, and have complete control of their inventory.
EHS-BSP shall assess special circumstances on a case by case basis upon PI request. Federal agencies retain the authority to, without prior notification, inspect and copy or request the submission of documentation.
Upon discovery of the theft or loss of select toxins, laboratory personnel must immediately notify the PI and EHS-BSP. EHS-BSP Deputy Director and SAT program manager shall notify appropriate agencies. Thefts or losses must be reported even if the select toxin is subsequently recovered or the responsible parties are identified.