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

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TABLE 1 PHYSICAL INACTIVATION OF SELECTED TOXINS TOXIN STEAM AUTOCLAVE DRY HEAT (10 MIN) FREEZE- THAW GAMMA IRRADIATION Botulinum neurotoxin Yes a > 100ºC b No c Incomplete d Staphylococcal Yes e > 100ºC; refolds f No g Incomplete h Enterotoxin Ricin Yes i > 100ºC i No j Incomplete k Microcystin No l > 260ºC m No n ND Saxitoxin No l > 260ºC m No n ND Palytoxin No l > 260ºC m No n ND Tetrodotoxin No l > 260ºC m No n ND T-2 mycotoxin No l > 815ºC m No n ND Brevetoxin (PbTx-2) No l > 815ºC m No n ND Table 1 Notes: ND indicates “not determined” from available decontamination literature. a Steam autoclaving should be at >121°C for 1 h. For volumes larger than 1 liter, especially those containing Clostridium botulinum spores, autoclave at >121°C for 2 h to ensure that sufficient heat has penetrated to kill all spores. 8,9 b Exposure to 100°C for 10 min. inactivates BoNT. Heat denaturation of BoNT as a function of time is biphasic with most of the activity destroyed relatively rapidly, but with some residual toxin (e.g., 1-5%) inactivated much more slowly. 10 c Measured using BoNT serotype A at -20°C in food matrices at pH 4.1-6.2 over a period of 180 days. 11 d Measured using BoNT serotypes A and B with gamma irradiation from a 60 Co source. 12,13 e Protracted steam autoclaving, similar to that described for BoNT, followed by incineration is recommended for disposal of SE-contaminated materials. f Inactivation may not be complete depending upon the extent of toxin re-folding after denaturation. Biological activity of SE can be retained despite heat and pressure treatment routinely used in canned food product processing. 14 g SE toxins are resistant to degradation from freezing, chilling or storage at ambient temperature. 15 Active SEB in the freeze-dried state can be stored for years. h References 15,16 I Dry heat of >100ºC for 60 min in an ashing oven or steam autoclave treatment at >121ºC for 1 h reduced the activity of pure ricin by >99%. 17 Heat inactivation of impure toxin preparations (e.g. crude ricin plant extracts) may vary. Heat-denatured ricin can undergo limited refolding (
k Irradiation causes a dose-dependent loss of activity for aqueous solutions of ricin, but complete inactivation is difficult to achieve; 75 MRad reduced activity 90%, but complete inactivation was not achieved even at 100 MRad. 18 Gamma irradiation from a laboratory 60 Co source can be used to partially inactivate aqueous solutions of ricin, but dried ricin powders are significantly resistant to inactivation by this method. l Autoclaving with 17 lb pressure (121-132 o C) for 30 min failed to inactivate LMW toxins. 17,19 All burnable waste from LMW toxins should be incinerated at temperatures in excess of 815°C (1,500 o F). m Toxin solutions were dried at 150 o C in a crucible, placed in an ashing oven at various temperatures for either 10 or 30 min, reconstituted and tested for concentration and/or activity; tabulated values are temperatures exceeding those required to achieve 99% toxin inactivation. 17 n LMW toxins are generally very resistant to temperature fluctuations and can be stored in the freeze-dried state for years and retain toxicity. TABLE 2 CHEMICAL INACTIVATION OF SELECTED TOXINS TOXIN NAOCL (30 MIN) NAOH (30 MIN) NAOCL + NAOH (30 MIN) OZONE TREATMENT Botulinum > 0.1% a >0.25 N ND Yes b neurotoxin Staphylococcal > 0.5% c >0.25 N ND ND enterotoxin Ricin > 1.0% d ND >0.1% + 0.25N e ND Saxitoxin ≥ 0.1% e ND 0.25% + 0.25N e ND Palytoxin ≥ 0.1% e ND 0.25% + 0.25N e ND Microcystin ≥ 0.5% e ND 0.25% + 0.25N e ND Tetrodotoxin ≥ 0.5% e ND 0.25% + 0.25N e ND T-2 mycotoxin ≥ 2.5% e,f ND 0.25% + 0.25N e ND Brevetoxin (PbTx-2) ≥ 2.5% e,f ND 0.25% + 0.25N e ND Table 2 Notes: ND indicates “not determined” from available decontamination literature. a Solutions of NaOCl (≥0.1%) or NaOH (>0.25 N) for 30 min inactivate BoNT and are recommended for decontaminating work surfaces and spills of C. botulinum or BoNT. Chlorine at a concentration of 0.3-0.5 mg/L as a solution of hypochlorite rapidly inactivates BoNT (serotypes B or E tested) in water. 20 Chlorine dioxide inactivates BoNT, but chloramine is less effective. 21 b Ozone (>2 mg/L) or powdered activated charcoal treatment also completely inactivate BoNT (serotypes A, B tested) in water under defined condition. 20,22 c SEB is inactivated with 0.5% hypochlorite for 10-15 mi. 23 d Ricin is inactivated by a 30 min exposure to concentrations of NaOCl ranging from 0.1- 2.5%, or by a mixture of 0.25% NaOCl plus 0.25 N NaOH. 17 In general, solutions of 1.0% NaOCl are effective for decontamination of ricin from laboratory surfaces, equipment, animal cages, or small spills. e The minimal effective concentration of NaOCl was dependent on toxin and contact time; all LMW toxins tested were inactivated at least 99% by treatment with 2.5% NaOCl, or with a combination of 0.25% NaOCl and 0.25N NaOH. 17 411
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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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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 Proper maintenance of ca
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Appendix A declined. However, in ma
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Appendix A It is possible to exhaus
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Appendix A 4. The Class II, Type A2
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Appendix A prohibited. Furthermore,
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Appendix A plastic wrappers, pipett
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Appendix A touch-plate microburners
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Appendix A SECTION VI FACILITY AND
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Appendix A Development of Containme
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Appendix A C. Airflow Smoke Pattern
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Appendix A TABLES Table 1. Selectio
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Appendix A Table 3. Field Performan
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Appendix A FIGURES Figure 1. HEPA f
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Appendix A Figure 3. The Class II,
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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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Page 393 and 394: Appendix D REFERENCES 1. Hess WR. A
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Page 403 and 404: Appendix H Working with Human, NHP
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credits bmbl December 7 '06.doc - Central Michigan University

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