Toxins of Biological Origin

bacteria snake fungi corn puffer fish

Biological toxins are produced by certain bacteria, fungi, protozoa, plants, reptiles, amphibians, fish, echinoderma (spiny urchins and starfish), mollusks, and insects.

Safety and Health Considerations For Conducting Work With Biological Toxins (.pdf article)

The EH&S Biosafety Office regulates the possession, use, and transfer of unfractionated mixtures and purified preparations of biological toxins with a mammalian LD₅₀ of ≤ 100 ug/kg body weight, as well as the organisms, both natural and recombinant, which produce these biological toxins. These are called “Acute Toxins”. Registration forms can be found at http://www.ehs.ufl.edu/Bio/default.asp

The following table from the UF EH&S Biological Safety Manual lists LD₅₀ values for some biological toxins.

Toxins not on this list may still require registration. For more information, please contact the Biosafety Office at 392-1591.

Toxin	LD50 (ug/kg)*
Abrin	0.7
Aerolysin	7.0
Botulinin toxin A	0.0012
Botulinin toxin B	0.0012
Botulinin toxin C1	0.0011
Botulinin toxin C2	0.0012
Botulinin toxin D	0.0004
Botulinin toxin E	0.0011
Botulinin toxin F	0.0025
b-bungarotoxin	14.0
Caeruleotoxin	53
Cereolysin	40-80
Cholera toxin	250
Clostridium difficile enterotoxin A	0.5
Clostridium difficile cytotoxin B	220
Clostridium perfringens lecithinase	3
Clostridium perfringens kappa toxin	1500
Clostridium perfringens perfringolysin O	13-16
Clostridium perfringens enterotoxin	81
Clostridium perfringens beta toxin	0.4
Clostridium perfringens delta toxin	5
Clostridium perfringens epsilon toxin	0.1
Conotoxin	12-30
Crotoxin	82
Diphtheria toxin	0.1
Listeriolysin	3-12
Leucocidin	50
Modeccin	1-10
Nematocyst toxins	33-70
Notexin	25
Pertussis toxin	15
Pneumolysin	1.5
Pseudomonas aeruginosa toxin A	3
Ricin	2.7
Saxitoxin	8
Shiga toxin	20
Shigella dysenteriae neurotoxin	1.3
Streptolysin O	8
Staphylococcus enterotoxin B	25
Staphylococcus enterotoxin F	2-10
Streptolysin S	25
Taipoxin	2
Tetanus toxin	0.001
Tetrodotoxin	8
Viscumin	2.4-80
Volkensin	1.4
Yersinia pestis murine toxin	10

*Please note that the LD50 values are from a number of sources (see below). For
specifics on route of application (i.v., i.p., s.c.), animal used, and variations on
the listed toxins, please go to the references listed below.

Reference:
1. Gill, D. Michael; 1982; Bacterial toxins: a table of lethal amounts;
Microbiological Reviews; 46: 86-94
2. Stirpe, F.; Luigi Barbieri; Maria Giulia Battelli, Marco Soria and Douglas
A. Lappi; 1992; Ribosome-inactivating proteins from plants: present status
and future prospects; Biotechnology; 10: 405-412
3. Registry of toxic effects of chemical substances (RTECS): comprehensive
guide to the RTECS. 1997. Doris V. Sweet, ed., U.S. Dept of Health and
Human Services, Public Health Service, Centers for Disease Control and
Prevention, National Institute for Occupational Safety and Health;
Cincinnati, Ohio

More Examples of Biological Toxins Which May Require Registration
Aflatoxins Amanitin Amphibian venoms Anatoxin A Anthrax toxin Aspergillus sp toxins Bacillus sp. toxins - all Bordetella sp. toxins Botulinum toxins - all Brevetoxins Bungarotoxins Cardiotoxin Charybdotoxin Cholera toxins - all Ciguatera toxin Clostridia species toxins - all Cluepeotoxins Cobra venous and all derived toxins Cobratoxin Conotoxins - all Crotamine Dendrodotoxins Dinoflagellate neurotoxins Diphtheria toxins Domoic acid DTX-1 (Dinophysistoxin-1) Echinoderm venoms - all Endotoxins - all Enterobacteriaciae toxins - all Enterotoxins - all Escherichia coli toxins - all Exotoxin A Fish venoms - all Fusarium sp. toxins Gliotoxin Joco Spider Toxin JSTX-3 Lappaconitines	Leiurotoxins Lipid A - all types Lipopolysaccharides from all species Maitotoxin Medamine Microcystins Mojave toxin Mycotoxins - all Myotoxins Neurotoxins - all Notexin Nodularin Ochratoxin Palytoxin Paradoxin Pertussis toxins - all Phalloidin Psilocybine Pseudomonas sp. toxins Reptile venoms - all Resiniferatoxin Ricin toxins - all Sapintoxin Sarafotoxin Saxitoxin Short Neurotoxins Snake venoms - all Stable toxins Staphylococcus sp. toxins Streptonigrin Taipoxin Tetanus toxins - all Tetrodotoxins - all Textilotoxin Thymeleatoxin Tinyatoxin Toxin II - all types

Toxins Classified as Select Agents

Some biological toxins are classified by the Federal Government as Select Agents (http://www.cdc.gov/od/sap/) due to their potential to pose a severe threat to public health and safety. Possession, use, and transfer of these toxins is highly regulated. A complete list can be found at http://www.cdc.gov/od/sap/docs/salist.pdf

In small quantities, some of these toxins are exempt from select agent registration. See the table below. Note however that the possession, use, or transfer of ANY select agent toxin, IN ANY QUANTITY, must be registered with the EH&S Biosafety Office. Again, forms can be found at http://www.ehs.ufl.edu/Bio/default.asp. Do not e-mail or fax select agent registrations to EH&S, hand-deliver or mail these registrations instead.

Exempt Amounts Select Agent Toxins Permissible Per Principal Investigator

HHS (CDC-listed) Toxins

Amount

Abrin

Toxic Hazard Label

100 mg

Conotoxin

100 mg

Diacetoxyscirpenol (DAS)

1000 mg

Ricin

100 mg

Saxitoxin

100 mg

Shiga-like ribosome inactivating proteins

100 mg

Tetrodotoxin

100 mg

HHS/USDA Overlap Toxins

Amount

Botulinum neurotoxins

0.5 mg

Staphylococcal enterotoxins

5.0 mg

Clostridium perfringens epsilon toxin

100 mg

Shigatoxin

100 mg

T-2 toxin

1000 mg

Working with and Disposing of Biological Toxins

Because they can be extremely hazardous, even in minute quantities, biological toxins require strict safeguards against their inhalation, absorption through skin or mucous membranes (typically due to a splash), ingestion, or percutaneous injury. Guidelines for the use of biological toxins can be found at

http://www.cdc.gov/od/ohs/biosfty/bmbl4/b4ai.htm

Key points of the guidelines are:

1. Written safety protocols to cover the use of the specific toxin(s) in use

2. Security measures in place to protect against unauthorized access to toxin(s)

3. Inventory control system in place; all entries in a hardbound book, in ink

4. Written plan for toxin-related emergencies (spill, exposure, etc) posted

5. BSL-2 or BSL-3 containment and practices in use

Specific inactivation and disposal requirements are in place for acute biological toxins. Some toxins are quite resistant to conventional methods of inactivation. These agents cannot be simply placed in the biomedical waste or picked up by EH&S Hazardous Waste Services.

Adapted from the University of Pennsylvania EH&S website:

Toxins may be destroyed by several methods as shown in the table below. Some toxins are inactivated by autoclaving for one hour at 121°C. Others are inactivated by exposure to sodium hypochlorite and/or sodium hydroxide.

A. Chemical destruction of toxins:

When using sodium hypochlorite and / or sodium hydroxide to destroy toxin, the procedure(s) must be performed in a laboratory fume hood or a biological safety cabinet. At a minimum, personal protective equipment for all procedures should include:

Long sleeved protective clothing (lab coat, gown)
Gloves and eye protection

If the toxin is classified as a select agent, even in exempt amounts, notify EH&S prior to destruction of the agent.
Work in a fume hood or biosafety cabinet with the sash at the lowest reasonable sash height for safe and effective work.
Place plastic backed absorbent paper (bench diaper) on the work surface of the fume hood or biosafety cabinet.
CAREFULLY put the Select Agent toxin into solution in the primary container. DO NOT USE A GLASS CONTAINER.
Place the primary container in a secondary container, such as a beaker or rack.
Slowly dispense an equal volume of the concentrations of sodium hypochlorite and/or sodium hydroxide designated in table 1 below into the primary container of toxin solution to be destroyed.
Do not replace the cap on primary container.
Place a “WARNING / DO NOT USE” sign on the hood/cabinet.
Allow a minimum 60 minutes exposure time. (See table 1 below for additional exposure time recommendations.)
Document the destruction of the toxin in the laboratory inventory logbook.
Secure the cap on the primary container. DOUBLE BAG the material in zip-lock plastic bags and label it “Inactivated/denatured (TOXIN NAME)”.
Contact EH&S for disposal as hazardous waste.

B. Steam Sterilization (Autoclaving) of Toxins

If acceptable as a method in table 1 below, destroy toxins by autoclaving them using the procedure outlined below:

If the toxin is classified as a select agent, even in exempt amounts, notify EH&S prior to destruction of the agent.
In a fume hood or biological safety cabinet, loosen the cap of the primary toxin container to allow steam penetration.
Place the primary container into a secondary biohazard sharps container.
Place the sharps container in a loosely closed biohazard bag.
Place the bag in a autoclavable pan.
Autoclave at 121° C for 1 hour on liquid cycle (slow exhaust).
Document the destruction of the toxin the laboratory inventory logbook.
After autoclaving, allow time for materials to cool before handling.
Discard the biobag and its containers as biological waste.

DO NOT use steam sterilization for destruction of any of the low molecular weight toxins (i.e. mycotoxins, marine and reptile venoms).

All waste from toxins that is not disposed as infectious waste must be collected by EH&S for disposal as hazardous waste. Call 392-1591.

Toxins classified as select agents, even if in exempt amounts, require that someone from the Biosafety Office observe and document the destruction of these agents. Please call our office (329-1591) to schedule an “observed destruct”.

Table 1
Inactivation Procedures for Selected Toxins

Allow at least a 60-minute chemical contact time for complete inactivation of toxin. Any procedure labeled “yes” is an approved procedure for inactivation of the toxin specified.

Toxin	Autoclave (1 hour @ 121° C, liquid exhaust)	2.5% NaOCL + 0.25 N NaOH	1.0% NaOCl	2.5% NaOCl
Abrin ⁽¹⁾⁽⁸⁾	Yes	N/A	N/A	N/A
Botulinum Neurotoxin ^{(1) (7)}	Yes	Yes	Yes	Yes
Clostridium perfringens epsilon toxin ⁽²⁾	Yes	N/A	N/A	N/A
Conotoxin⁽³⁾	CALL EH&S
Diacetoxyscirpenol⁽⁵⁾	No	Yes	No	Yes (3-5%)
Ricin ⁽¹⁾⁽⁷⁾	Yes	Yes	Yes	Yes
Saxitoxin ⁽¹⁾⁽⁷⁾	No	Yes	Yes	Yes
Shigatoxin & Shiga-like ribosome inactivating proteins⁽⁴⁾	Yes	Yes	Yes	Yes
Staphylococcal Enterotoxins ⁽¹⁾⁽⁷⁾	Yes	Yes	Yes	Yes
Tetrodotoxin ⁽¹⁾⁽⁷⁾	No	Yes	Yes	Yes
T-2 Toxin ^(1)(6)(5)	No	Yes	No	No

Wannemacher R.W. 1989. Procedures for Inactivation and Safety Containment of Toxins. Proc. Symposium on Agents of Biological Origin, U.S. Army Research, Dev. and Engineering Center, Aberdeen proving Ground, MD. pp. 115-122
Factsheets on Chemical and Biological Warfare, http://www.cbwinfo.com/Biological/Toxins/Cper.html
Factsheets on Chemical and Biological Warfare, http://www.cbwinfo.com/Biological/Toxins/Conotox.html
Factsheets on Chemical and Biological Warfare, http://www.cbwinfo.com/Biological/Toxins/Verotox.html
Factsheets on Chemical and Biological Warfare, http://www.cbwinfo.com/Biological/Toxins/mycotoxins.html
For complete inactivation of T-2 mycotoxin extend exposure time for liquid samples, spills, and non-burnable waste in 2.5% sodium hypoclorite and 0.25 N sodium hydroxide to 4 hr. Expose cages/bedding from animals exposed to T-2 mycotoxin to 0.25% sodium hypochlorite and 0.025 N sodium hydroxide for 4 hrs.
For inactivation of saxitoxin, tetrodotoxin, ricin, botulinum toxin, or staphylococcal enterotoxins, expose work surfaces, solutions, equipment, animal cages, spills to 10% sodium hypochlorite for 60 minutes.
http://www.inchem.org/documents/pims/plant/abruspre.htm

Please don’t hesitate to contact the EH&S Biosafety Office with any questions regarding biological toxins.

You can reach us at: 352-392-1591, E-mail: bso@ehs.ufl.edu