credits bmbl December 7 '06.doc - Central Michigan University

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Agent Summary Statements – Toxins formed BoNT is the primary concern for laboratory workers. Two of the most significant hazards in working with BoNT or growing C. botulinum cultures are unintentional aerosol generation, especially during centrifugation, and accidental needle-stick. Although BoNT does not penetrate intact skin, proteins can be absorbed through broken or lacerated skin and, therefore, BoNT samples or contaminated material should be handled with gloves. Workers in diagnostic laboratories should be aware that toxin-producing C. botulinum or its spores can be stable for weeks or longer in a variety of food products, clinical samples (e.g., serum, feces) and environmental samples (e.g. soil). Stability of the toxin itself will depend upon the sterility, temperature, pH and ionic strength of the sample matrix, but useful comparative data are available from the food industry. BoNT retains its activity for long periods of time (at least 6-12 months) in a variety of frozen foods, especially under acidic conditions (pH 4.5-5.0) and/or high ionic strength, but the toxin is readily inactivated by heating. 5 A documented incident of laboratory intoxication with BoNT occurred in workers who were performing necropsies on animals that had been exposed 24 h earlier to aerosolized BoNT serotype A; the laboratory workers presumably inhaled aerosols generated from the animal fur. The intoxications were relatively mild, and all affected individuals recovered after a week of hospitalization. 6 Despite the low incidence of laboratory-associated botulism, the remarkable toxicity of BoNT necessitates that laboratory workers exercise caution during all experimental procedures. Containment Recommendations BSL-2 practices, containment equipment, and facilities are recommended for routine dilutions, titrations or diagnostic studies with materials known to contain or have the potential to contain BoNT. Additional primary containment and personnel precautions, such as those recommended for BSL-3, should be implemented for activities with a high potential for aerosol or droplet production, or for those requiring routine handling of larger quantities of toxin. Non-immunized personnel should be discouraged from entering the laboratory when BoNT is in use until after the toxin and all work surfaces have been decontaminated. Purified preparations of toxin components, e.g. isolated BoNT “light chains” or “heavy chains,” should be handled as if contaminated with holotoxin unless proven otherwise by toxicity bioassays. SPECIAL ISSUES Vaccines A pentavalent (A, B, C, D and E) botulinum toxoid vaccine (PBT) is available through the CDC as an IND. Vaccination is recommended for all personnel working in direct contact with cultures of C. botulinum or stock solutions of BoNT. Due to a possible decline in the immunogenicity of available PBT stocks for some toxin serotypes, the 286
Agent Summary Statements – Toxins immunization schedule for the PBT recently has been modified to require injections at 0, 2, 12, and 24 weeks, followed by a booster at 12 months and annual boosters thereafter. Passive Immunization Experimental studies in NHP have demonstrated that administration of purified equine or human immune globulin may also confer passive protection against inhalation or injection of toxin. 7 Select Agent C. botulinum toxin is a Select Agent requiring registration with CDC and/or USDA for possession, use, storage and/or transfer. See Appendix F for additional information. Transfer of Agent Importation of this agent may require CDC and/or USDA importation permits. Domestic transport of this agent may require a permit from USDA/APHIS/VS. A DoC permit may be required for the export of this agent to another country. See Appendix C for additional information. Agent: Staphylococcal enterotoxins Staphylococcal enterotoxins (SE) are a group of closely related extracellular protein toxins of 23 to 29 kD molecular weight that are produced by distinct gene clusters found in a wide variety of S. aureus strains. 8,9 SE belong to a large family of homologous pyrogenic exotoxins from staphylococci, streptococci and mycoplasma which are capable of causing a range of illnesses in man through pathological amplification of the normal T- cell receptor response, cytokine/lymphokine release, immunosuppression and endotoxic shock. 9,10 SE serotype A (SEA) is a common cause of severe gastroenteritis in humans. 11 It has been estimated from accidental food poisoning that exposure to as little as 0.05 to 1 µg SEA by the gastric route causes incapacitating illness. 12-15 Comparative human toxicity for different serotypes of SE is largely unknown, but human volunteers exposed to 20-25 µg SE serotype B (SEB) in distilled water experienced enteritis similar to that caused by SEA. 16 SE are highly toxic by intravenous and inhalation routes of exposure. By inference from accidental exposure of laboratory workers and controlled experiments with NHP, it has been estimated that inhalation of less than 1 ng/kg SEB can incapacitate more than 50% of exposed humans, and that the inhalation LD 50 in humans may be as low as 20 ng/kg SEB. 17 Exposure of mucous membranes to SE in a laboratory setting has been reported to cause incapacitating gastrointestinal symptoms, conjunctivitis and localized cutaneous swelling. 18 287
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Biosafety in Microbiological and Bi
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SECTION VIII-H: Prion Diseases 299
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Forward Biosafety in Microbiologica
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Editors: L. Casey Chosewood, MD Dir
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Guest Editors: Matthew J. Arduino,
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Charles B. Millard, PhD Lieutenant
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Robert Bull Karen B. Byers Jane Cap
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Thomas L. Miller Douglas M. Moore M
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Robert Yarchoan Uri Yokel Lisa Youn
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Introduction laboratories chose not
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Introduction RISK CRITERIA FOR ESTA
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Introduction BMBL includes an impor
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Section II Biological Risk Assessme
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Biological Risk Assessment informat
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Biological Risk Assessment The NIH
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Biological Risk Assessment effectiv
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Biological Risk Assessment Third, m
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Biological Risk Assessment 14. Oxma
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Principles of Biosafety Safety equi
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Principles of Biosafety Often an in
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Principles of Biosafety Four standa
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Principles of Biosafety 4. Centers
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Laboratory Biosafety Level Criteria
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Section V Vertebrate Animal Biosafe
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Vertebrate Animal Biosafety Level C
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TABLE 1 SUMMARY OF RECOMMENDED BIOS
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Principles of Laboratory Biosecurit
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Occupational Health and Immunoproph
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Agent Summary Statements - Bacteria
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• Section VIII-B: Fungal Agents A
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Agent Summary Statements - Fungal A
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• Section VIII-C: Parasitic Agent
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Agent Summary Statements - Parasiti
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• Section VIII-D: Rickettsial Age
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Agent Summary Statements - Ricketts
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Agent Summary Statements - Ricketts
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• Section VIII-E: Viral Agents Ag
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Agent Summary Statements - Viral Ag
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Appendix A Figure 8. The Class III
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Appendix A Figure 9B. The vertical
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Appendix A Figure 11. A typical lay
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Appendix A REFERENCES 1. Centers fo
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Appendix B Decontamination and Disi
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Appendix B surfaces are contaminate
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Appendix B TABLE 1 DESCENDING ORDER
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Appendix B dioxide gas exits the ge
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Appendix B d The effectiveness of a
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Appendix C Transportation of Infect
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Appendix C 1600 Clifton Road, N.E.,
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Appendix C disease in humans or in
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Appendix C Biological specimen, Cat
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Appendix D Camelpox virus b a, b, c
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Appendix D SPECIAL ISSUES The impor
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Appendix D required by the USDA and
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Appendix D depression, anorexia, an
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Appendix D laboratory with enhancem
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Appendix D five day period of conta
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Appendix D virus does not cause any
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Appendix D Spring Viremia of Carp V
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Appendix D REFERENCES 1. Hess WR. A
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Appendix D 37. Alexander DJ. Newcas
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Appendix E Arthropod Containment Gu
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Appendix F Select Agents and Toxins
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Appendix G IPM issues and requireme
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Appendix H Working with Human, NHP
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Appendix I Guidelines for Work with
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decontaminated periodically, for ex
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0.25N, and/or sodium hypochlorite (
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k Irradiation causes a dose-depende
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7. Morin R, Kozlovac J. Biological
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Appendix J NIH Oversight of Researc
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Appendix K Resources for Informatio
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Appendix K E-mail: http://www2.gsu.
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Appendix K and http://www.who.int/c
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Appendix L HEPA High Efficiency Par
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