: : AD ROTE Project No. _1M4~65710-D049 TECOM Project No. 2-CO-210-049-002 DPG Document No. _DPG-TR-82-005 JOINT CB TECHNICAL DATA SOURCE BOOK (U) VOLUME VI. TOXIN AGENTS (U) PART ONE: BOTULINUM TOXIN (U) JUNE 1982 U.S. ARMY DUGWAY PROVING GROUND Dugway, Utah 84022 Copy __ 2 of 112 Copies ppg 82-227UNCLASSIFIED REPORT DOCUMENTATION PAGE Berne Comet crn eae TECOM No. 2-CO-210-049-002 | TITLE fave Bone? TIVE OF REPORT & PERIOD COVERED Joint CB Technical Data Source Book, Source Book/to 1972 Volume VI, Toxin Agents, Part One, [i eRFoRmNG ORG, REPORT NUMBER — Botulinum Toxin (U) DPG-TR-82~005 TROFHORTY '7-CONTRACT OF GRANT NUMBERT DAADO9-69-C-0078 | I” BERFORMING ORGANIZATION WANE AHO AOORERS 7 See US Army Dugway Proving Ground, STEDP-MT-DA-S Dugway, UT 84022 and | am4-65710-D049 GEOMET, INC TT CONTROLLING OFFICE WANE AND AODRESE | REPORT Date US Army Dugway Proving Ground sume 282 Dugway, UT 84022 eee oe 62 Ta" WOWITSAINS AGENGY WAME ROGREGHI afarantFos Contrliing Oiica) | 18 SECURITY CAGE, (al Ble FoporD _: 4 [Sa GECLASHETEATION/OOWNGRRGTRG— Review 30 Jun 02 TE GISTRIBUTION STATEMENT (ol ie Reprty Distribution limited to US Government agencies only; Test and Evaluation (June 1982). Other requests for this document must be referred to: Commander, US Army Dugway Proving Ground, ATIN: STEDP-MT-DA-S, Dugway, ut 84022. TF BISTRIGUTION STATEMENT (ol the Tat mitered Ih Black 36, 1 alacant fom Roper) { Antigens Biologice: aeros Botulinur toxin Toxin defense Dose/dosages TES ABETRACT (Coin ar roveien 0s T reconensy ad Idnlly BY SISGE Rambo) | (U) Part One of Volume VI of the JOINT CB TECHNICAL presents c nce levels derived from laboratory | and cham! Data are presented that are for botulinum ailable se with the a cion, where c ‘tulinum toxin ara presented. DD TSIM, M73. commen ors wow ss 1s osousTe - UNCLASSIFIEDUNCLASSIFIED FOREWORD (U) This document was prepared in compliance with Departuent of the Ary letter, subject: Joint Contact Point for Chemical-Biological (CB) Field Test Data, 10 March 1967, which directed Deseret Test Center (DTC) to publish and maintain a Joint CB Technical Data Source Book. (U) The Source Book is organized into a series of volumes, each addressing an identifiable area of information related to the analysis of CB weapons and defensive systems. Areas include agents, general models, weapon systems, assay and data reduction procedures, chemical simulants and biological nonpathogens, and knowledge deficiencies. (WV) The Source Book is designed to be used by the research and developnent community as input into defensive techniques and defensive devices. (W) Portions of the information contained in the Botulinum Toxin Source Book were compiled by GEOMET, Inc., under Contract DAADO9-69-C-0078, with the remainder by analytical personnel of Deseret Test Center and US Army Dugway Proving Ground. All material has been subjected to review and coordination by selected members of the CB community. The conscientious efforts expended by those individuals to improve the quality of the final product is gratefully acknowledged and appreciated. (U) The information in this volume was initially gathered in 1972, and generally represents the state of the art as of that date. This published volume replaces a draft entitled JOINT CB TECHNICAL DATA SOURCE BOOK, Volume VI, Toxin Agents, Part Two: AGENT XB (0), July 1977, DTIC No 80C027270, which should be destroyed. 4 UNCLASSIFIEDpremied Page La FOREWORD LIST OF TABLES 2. ee 3 . vit LtstJOnriGunes| en : toe at 1-1 Introduction . ee 1-2 Agent Description : er 1-3. Phystological Response . eee QF 1-4 Source Model and Paraneters oe 1-5 Atmospheric Transport Model 6o20020 ; 71. 1-6 Detection-Protection po oo8 CHAPTER : DUCTION 22h 2-1 General . Geog po ooo ee aes 2-2 Description of Botulinum Toxic. “re A (Agent XR). + + = a 2-3 Model Requirements . ee 2-4 Delivery Systems... ee CHAPTER 3. PHYSTOLC 3-1 General 5 a os 3-2 Mechanism of action. 6 2. | | Le . dl | 3-3 Casualey Probability . ee | 3-4 Tine to Casualty (Respiratory oS 3-5 Duration and Severity of Casue oo 7 3-6 Physiological Countermeasures E CHAPTER 4. souRc f _ t Laat os 1 7 MCDEL_AND PARAMETERS 5 ' i : Sa tis UNCLASS-FTEDUNCLASSIFIED ‘TABLE OF CONTENTS (Cont'd) Page I CHAPTER 6. DELIVERY MODEL AND PARAMETERS Rr 1 i fp pelivery Errors es OF ' CHAPTER 7. DEFENSE MODELS AND PARAMETERS i 4-2. Protection Model Parameters. - + + + + st 5" * 7-4 4 ie, eet ee ioe mode! racsmetere tt 7-5 , CHAPTER 8. IDENTIFICATION OF KNOWLEDGE GAPS 8-2 Models and Parameters. © + + + © 7 tt ot Roy eee oo APPENDIX B. Glossary and Abbreviations =. - + + DIX C. Distribution List 6 6 6 ee ee ttt OF34 4-2 4-3 Inhalation LDsq for XR for Variou corresponding LDso and LCtso for Estimated Parameter Values for Ti= co Protection Afforded by Pentavale:= = Aerosol Challenge with XR. potulism Mortality - Antitoxin Untreated Cases s+ + + + Toxicity of Partially Purified T-x from a Suspension at pH 3.9 aft Constant Temperature - + + Toxicity of Partially Purified from a Solution at pil 6.5 after Constant Temperature - + + ‘Toxicity of Pure Toxin Lyophiliz at pH 3.0 after Storage at Const Toxicity of Pure Toxin. Lyophil: at pH 6.5 after Storage at Const y of Dry XR During Storaz= lean Errors from Observed Toxic ing Quadratic Decay Constar: Srorage Temperatures Mean Errors from Observed Toxic=2: near Decay Constants for XR @ Temperatures Fenaturation . ++ Toxicity of XR with Time in wuids and Storage Temper of Toxicity Stability y the data in Table 374; aye nese aerictedl care lGrom) at tzetdon ag 21 different sources. (u) The only 51 sooner the antitoxin is rable 3-4, (U) Botulism Mortality ~ Anticoxin ‘Treated Versus | Untreated Casest UNCLASSIFIED Wuuber of Cases a Treatment survived Died ‘Total Treated 25 9 34 untreated 5 37 42 Thmong those treated, 74 percent survived, while anon those not q treated, only 12 percent survived.qable 4-1. (WU) Toxicity of P Temperature. UNCLASSIFIED artially Purif Suspension at pH 3.9 after St Toxieity (10° MIPLDso/mt) ‘Storage Temperature fed Toxin Lyophilized from rage at Constant orage Time (months) -18 °C 5 oC 25 0c © (control) ay 358 35° 23 23 18 23 19 25 23 23 27 36 33 30 34 33 31 39 28 25 35 33 33 43 53 36 2 32 28 22 l *pased on measured toxicity of 70 for suspension (dil prior to freezing. uted 1 to 2)a. (U) Point Sour agent disseminated by explosi time duration) is source strength for a point source of generative mechanisms (with short VG, ® (4.8) where Q = source strength of the agent cloud formed by the munition, MEPLDso Vv = quantity of agent in the munition, mt or g G = concentration of the agent fill, MIPLDy, per m or g 2 = efficiency of the munition in converting agent £111 into aerosol particles in the proper size range, dimensionless b. (U) Line Source. The source strength for 2 line source dissemin- ating device is EG, é (4.9) where q = source strength of the agent cloud per unit of line length, MIPLDs per m E = rate of agent emission from the device, mf/min or g/min S = speed of delivery vehicle carrying the disseminating device, a/min G, = concentration of the agent fill, MIPLD,,, per mi or g c. (U) Line of Point Sources. A line of relatively closely spaced point sources (bomblets or generators) may be treated as a line source when the effects to be considered are at a sufficient distance downwind where the point source clouds have merged. In this case, the source strength model is: nusber of point sources x Q Tine Tengeh ae (4.10)agent fii") very of aerosols below 5 ® = 9.3 (based © ein analysis pased on total prot: ‘The resuits tt concentration. intrations. (above) is as using a single agent a to extrapolate to other conce: of five muniti Figure 4-2) 02(U) The basic data were analyzed to determine biological and physical exponential decay constants for each of the 20-liter chamber tests. Where only two samples vere obtained from the cloud, the decay constant is given by k an SL (5.2) ae" where P, = toxicity (or Lowry protein concentration) of aerosol cloud at first sampling time 2 = toxicity (or Lowry protein concentration) of aerosol cloud at second sampling time At = time between samples. Where three or more samples were obtained, the decay constant was computed by linear regression between the natural logarithm of toxicity and time. ‘The computed decay constants are listed in Table 5-3.spiratory exposure 1s espiratory dose as 3 result © t) de any ey err f XK) BE) © q(t) here dg(t) = snhated dove retained up £0 tine & (ainutes), in wg for XE bf toxin within respiratory tract, This ts wntrd paraceter which has an assubed value of This value £5 based on data from a sampling ‘and retention characteristics Porton preimpinger/all glass R = retentivity an agent- order 1.0 for XR. systen simulating intake Of the human lung, t.¢-y {mpinger coubination X(t) = emeentration history of tndivideal's Location, tn ng min/or? for XR a(t) © beeathing Fate as 2 function of tine (related to physical activity), A/nin a(t) «masking function, whtch reflects Jegree of respiratory maetteE on afforded by the nash asa function of training, Protecr toed cask ceaign expressed as fractional exposure eee tey, varying bet=een zero and one ‘The breathing rate and usbing functine parameter values are given in Volune X (Ccceral Mouele), The only aceat-ortented paraneter is chat for tetentisite CR) hie, bas an assumed value of 1,0 for XR (100 per- cent retention). 7-3. Decontamination Mode! Param (&) Table 7-2 sumarires the agsesszent data for standard fleld decontaminaes relation #1 ehelr abs: ity te detoxify expected levels of dotulinun toxin, fable 2 presents a quantitative assessment of the effietency of various con entrath for the decontaminants in question agcvood srsenal!*) indicate thar effective 3 de accomplished with an alkaline Feast 2,500 parts ner illion af s given In Table 7-3 (U) Tnvestizators at decontamination of botulicz: toxin = hypochlorice siurry active chlorine atot UNCLASSIFIED i # t) Assessment Data of Field Decontaminants to Detoxify Botulinum Toxin. (2) Mouse Mortality Fraction* 2 Contact Contact # Lo mint (5 min) active | wa [is ot ¢ ¢ pecontaminant cntorine] @ 2], | Se | Ee | Se G Be] oa Sa ga ga §| $3 | 383 | $2 33 3] 82 | 26 | ae) 28 8 a 2 2 a 6S 3 63 Ss ak sh | ck ck StS (super tropical 15.5 |10.7 | ov10 | o/to | o/20| 0/20 bleaching powder (chlorinated lime)) HTH powder (high-test 35.0 |11.0 | 0/10 | o/10 | 0/20] 0/20 hypochlorite (calcium hypochlorite)) DS2 Liquid (diethyl- see [tr.o®| o710 | o/10 | 20/20]... enetriamine 69%; NaOH 2% methyl cellosolve 29%) 5330 powder (chloramide) | 29.5 | 2.6 | o/t0 | 9/10 | ... we xxCC, powder (chloranide)| 12.8 | 7.2 | 10/10 |... | oe - XXCC, fabric (standard 1.3 | 6.8¢| 10/10 |... | o. impregnated 8-1/2 oz. ay cotton sateen, 1x 6 in, piece) The control dilution of the XR suspension gave a mouse mortality fraction of 100 percent, The control dilution of the decontaminants had no apparent effect on mice. vpHl drops rapidly in contact with air because of carbonate formation. cFabric netted with gelatin-phosphate diluent. substantiate this. Both super tropical bleaching powder (STB) and high test hypochlorite (HTH) decontaminants were very effective in decontami- ' nating concentrations ranging from 1 to 35 percent with reaction times | as short as 5 minutes. Results with DS2 (69% diethylenetriarine, 2% NaOH, and 29% methyl cellosolve) and $330 (chloramide) were not as nclusive ac with the hypochlorites. While the test resulte indicated | some reactivity, the assessment of effectiveness in the fielé requires UNCLASSIFIED yUNCLASSIFIED 4. (WU) Qualitative Test Summary.) UNCLASSIFIED Concentration of|Concentration of [Concentration of Field Decontaminant | Decontaminant | Decontaminant Decontamination | 50%; Dry Ratio | 10%; Dry Ratio | 12; Dry Ratio Material of Decontaminant [of Decontaminant lof Decontaminant to XR, 100:1 | to XR, LO: to XR, 1:0 STB (super ‘tropical Good Good Good* bleaching powder (chlorinated lime)) HTH (high-test Good Good Good* hypochlorite (caleiun hypochlorite)) ps2 liquid Good Good Not recommended (diethylenetriamine 69%; NaOH 2%; methyl’ cellosolve 29%) $330 powder Good Not recoumended ee (chloramide) XXCC, powder Not recommended a ee (chloramide) XXCO, fabric Not recomended a (standard impregnated 8 1/2- ounce cotton sateen, 1 by 6 in. piece) I “This test was repeated, and the reaction time was reduced from 15 to 5 minutes. Results were the same (very good decontamination). additional testing. Results with XXCC3 impregnated fabric showed that the standard clothing impregnate had little or no reactivity with botulinum toxin. (U) Requirements for specialized deccntamination of equipment and surfaces contaminated with toxins (of which botulinum toxin is typical) led to the investigation of formaldehyde gas released from dry paraformaldehyde as a detoxifying agent. In assessing paraformaldehyde, botulinum toxin, type A, was used as both a liquid suspension (at a concentratic TPLDsq pet millilicer) and as a powder (at a concentratio: TPLDsq per gram)‘ * UNCLASSIFIED