http://ehs.utah.edu/research-safety/biosafety/protocol-review/biological-toxins-registration
Biological Toxins
Biological toxins are toxic substances produced by bacteria, fungi, protozoa, insects, animals, or
plants that have the capability of causing harmful effects when inhaled, ingested, injected or
absorbed. They may be transmitted via surface contact with contaminated object(s) and
subsequently spread to mucus membranes (eyes, nose, and mouth) and/or to open sores on
skin. Some biological toxins can be absorbed through intact skin, especially if solubilized in
substances such as dimethyl sulfoxide (DMSO). Accidental needle-stick is a mode of
transmission within research laboratories. Accidental ingestion of contaminated materials and
inhalation are other routes of transmission. While they cannot replicate and are not infectious
they can be extremely hazardous, even in minute quantities. The health effects of exposure can
vary greatly depending on the toxin, the amount, and the route of exposure, ranging from
minor (skin or eye irritation, headache, nausea) to severe (respiratory distress, muscle
weakness, seizures, death).
IBC Registration
The University of Utah Institutional Biosafety Committee (IBC) reviews registrations for work
with, possession of, use of, and transfer of acute biological toxins (mammalian LD50 <100 µg/kg
body weight) or toxins that fall under the Federal Select Agent Guidelines, as well as the
organisms, both natural and recombinant, which produce these toxins
The following is a list of toxins that are required to be registered with the IBC. However, the list
is not comprehensive and principal investigators can confirm that toxins they propose to work
with do not require IBC registration (LD50 >100 µg/kg body weight and not on Select Agent list)
by contacting the Biosafety Office (biosafety@ehs.utah.edu or 801-581-6590).
Abrin
Aflatoxin
Bacillus anthracis edema factor
Bacillus anthracis lethal toxin
Botulinum neurotoxins
Brevetoxin
Cholera toxin
Clostridium difficile toxin
Clostridium perfringens toxins
Conotoxins
Dendrotoxin (DTX)
Diacetoxyscirpenol (DAS)
Diphtheria toxin
Domoic acid
Pertussis toxin
Pfeisteria spp. toxin(s)
Ricin
Saxitoxin
Shiga-like ribosome inactivating proteins
Shigatoxin
Staphylococcal enterotoxins
T-2 toxin
Tetanus toxin
Tetrodotoxin (TTX)
The University of Utah Office of Environmental Health and Safety is migrating management and
tracking of laboratories and research to BioRAFT. PIs or their delegates working with acute
toxins must complete the General Laboratory Setup and Biological Registration Wizards.
Currently there is not a Toxin registration form. Please complete the Toxin Registration form
that can be downloaded below and add as a “Document” in BioRAFT.
Click here (https://ehs.utah.edu/research-safety/biosafety/bioraft) to access the University of Utah
BioRAFT page.
Downloads:
Biological Toxins Registration-BioRAFT form (link to Word Document)
The laboratory will be expected to maintain a current chemical hygiene plan (University of Utah
template can be found here (http://ehs.utah.edu/sites/default/files/research-safety/chemicalsafety/chemical-hygiene-plan/ehs-chemical-hygiene-plan-template.pdf)) and/or Standard
Operating Procedures (University of Utah template can be found here (link to BSL2 SOP)).
Laboratory Practices
Guidelines for working with biological toxins can be found in Appendix I of the Biosafety in
Microbiological and Biomedical Laboratories
(http://www.cdc.gov/biosafety/publications/bmbl5/index.htm). These are summarized below.
Routine operations with dilute toxin solutions are conducted using Biosafety Level 2 (BSL2)
practices and these must be detailed in the IBC protocol and will be verified during the
inspection by EHS staff prior to IBC approval. BSL2 Inspection checklists can be found here
(http://ehs.utah.edu/research-safety/biosafety/biosafety-laboratory-audits). All personnel
working with biological toxins or accessing a toxin laboratory must be trained in the theory and
practice of the toxins to be used, with special emphasis on the nature of the hazards associated
with laboratory operations and should be familiar with the signs and symptoms of toxin
exposure. This includes how to handle transfers of liquids containing toxin, where to place
waste solutions and contaminated materials or equipment, and how to decontaminate work
areas after routine operations, as well as after accidental spills. The worker must be reliable and
sufficiently adept at all required manipulations before being provided with toxin. Laboratory
work with toxins should be done only in designated rooms with controlled access and at predetermined bench areas. When toxins are in use, the room should be clearly posted: “Toxins in
Use—Authorized Personnel Only.”
Researchers working with a toxin should be vaccinated if a vaccine is available (e.g. diphtheria
toxin, tetanus toxin). Routine operations with dilute toxin solutions are conducted using BSL2
practices and facilities shall be used for activities involving biological toxins. These include:
 Biohazard signs and labels must be displayed in areas and on equipment where
biological toxins are used and stored. This includes, but is not limited to, laboratory
entrance doors, biological safety cabinets, chemical fume hoods, refrigerators, and
freezers.
 Use a biological safety cabinet (BSC) or a chemical fume hood for resuspension of
biological toxins or manipulations of stock solutions of toxins that can generate
aerosols, such as pipetting, harvesting, infecting cells, filling tubes/containers, and
opening sealed centrifuge canisters. When using an open-fronted fume hood or BSC,
workers should wear suitable laboratory PPE to protect the hands and arms, such as
laboratory coats, smocks, or coveralls and disposable gloves.
 Whenever possible, use needle-free techniques to resuspend biological toxins.
 If a quantity of powder-form toxin must be weighed, then the scale must be located in a
certified chemical fume hood.
 When conducting liquid transfers and other operations that pose a potential splash or
droplet hazard in an open-fronted hood or BSC, workers should wear safety glasses and
disposable facemask, or a face shield.
 Toxin should be removed from the hood or BSC only after the exterior of the closed
primary container has been decontaminated and placed in a clean secondary container.
Toxin solutions, especially concentrated stock solutions, should be transported in
leak/spill-proof secondary containers.
Emphasis must be placed on evaluating and modifying experimental procedures to eliminate
the possibility of inadvertent generation of toxin aerosols. Pressurized tubes or other
containers holding toxins should be opened in a BSC, chemical fume hood, or other ventilated
enclosure. Operations that expose toxin solutions to vacuum or pressure, for example
sterilization of toxin solutions by membrane filtration, should always be handled in this manner,
and the operator should also use appropriate respiratory protection. If vacuum lines are used
with toxin, they should be protected with a HEPA filter to prevent entry of toxins into the line.
Centrifugation of cultures or materials potentially containing toxins should only be performed
using sealed, thick-walled tubes in safety centrifuge cups or sealed rotors. The outside surfaces
of containers and rotors should be routinely cleaned before each use to prevent contamination
that may generate an aerosol. After centrifugation, the entire rotor assembly is taken from the
centrifuge to a BSC to open it and remove its tubes.
Experiments should be planned to eliminate or minimize work with dry toxin (e.g., freeze-dried
preparations). Unavoidable operations with dry toxin should only be undertaken with
appropriate respiratory protection and engineering controls.
Personal Protective Equipment (PPE)
Work with biological toxins shall be conducted using BSL2 PPE:
 Disposable gloves – consider the use of double gloves for enhanced protection. Ensure
your gloves are compatible with any solvent your toxin may be dissolved in.
 Lab coat or back-closing disposable gown
 Eye protection (safety glasses or goggles) is recommended, but not required when
working within a biosafety cabinet or a chemical fume hood.
An inventory control system should be in place to account for toxin use and disposition. If
toxins are stored in the laboratory, containers should be sealed, labeled, and secured to ensure
restricted access; refrigerators and other storage containers should be clearly labeled and
provide contact information for trained, responsible laboratory staff.
Decontamination and Spills
Toxin stability varies considerably outside of physiological conditions depending upon the
temperature, pH, ionic strength, availability of co-factors and other characteristics of the
surrounding matrix. Literature values for dry heat inactivation of toxins can be misleading due
to variations in experimental conditions, matrix composition, and experimental criteria for
assessing toxin activity. Moreover, inactivation is not always a linear function of heating time;
some protein toxins possess a capacity to re-fold and partially reverse inactivation caused by
heating. In addition, the conditions for denaturizing toxins in aqueous solutions are not
necessarily applicable for inactivating dry, powdered toxin preparations. Inactivation
procedures should not be assumed to be 100% effective without validation using specific toxin
bioassays.
Many toxins are susceptible to inactivation with dilute sodium hydroxide (NaOH) at
concentrations of 0.1-0.25N, and/or sodium hypochlorite (NaOCl) bleach solutions at
concentrations of 0.1-0.5% (w/v). Use freshly prepared bleach solutions for decontamination;
undiluted, commercially available bleach solutions typically contain 3-6% (w/v) NaOCl: a 1:10
dilution of bleach gives an appropriate concentration.
Exposures
Antitoxins are available for some biological toxins and immediate medical “first-aid”
interventions may help prevent or lessen the severity of the reaction. If you know or suspect a
biological toxin exposure.
 Irrigate the site of exposure
o If exposure was by needle stick or other route which breaks the skin, wash with
soap and water for 5-15 minutes and cover with a bandage.
o If exposure was by splash to eyes or mucus membranes, irrigate thoroughly for
15 minutes at an appropriate eye wash station.
 Report to your Laboratory Supervisor and the Biosafety Officer IMMEDIATELY. Seek
medical attention as outlined in your laboratory specific safety manual.
Export Controlled Toxins
Several biological toxins are restricted for export by the U.S. Department of Commerce and
require an export license prior to any shipment out of the U.S. Many of the toxins on the
export control list do not meet the definition of an acute toxin (LD50 <100 µg/kg body weight)
and do not require registration with the IBC but the Principal Investigator is responsible for
ensuring that they are in compliance with the export control laws and regulations. If you have
questions about export controls, please visit the Office of Sponsored Projects webpage
(http://osp.utah.edu/policies/export-controls.php) or contact OSP’s Export Control Officer,
Todd Nilsen.