oaoUPI

(-.0)

Dl!II __ ,. 19L8

I.e at ~teal 1I1ebol8~. rahbS..~ 1IIoculated aD .Ipril. 19, 19b8.

Be ~t.1ca at .1I1a1C1lI

1. twUcl8e r..ond at. 6100 ...... CD.., 9. 19118. L To tM t.t.1~, IIIftIIle ·'NdDI ~ 1d.th ..... , .. addId 1MO co

at tr.b·wdDac\1 ... IIIbt4t, a.r. C- rabld.') 8DI1.iI4P oc at n.ru. DIU ]..u.n.. 'lbe _tenal .. II1Dd w au. tile 1IP1z'ocbM- to • raw rr. tot. \luoa8 ad ~ t I 'I --It

3. n. ....... laa.. .. • _ apMd, riwKwt&ti (10. 1 Sd. c-trUup> 8R ... 10 t. S 1dD ....

4. IDtllerat.1ca or •

20 t1wldll wn ..... 14. anll'al. oc,...~ taotor of U. 1111_ .. .. ialoc_7 .. ....

C. PNparaUa aDd ~ of Pat.UatBt

Par .... pat1eat. tbe t .... 1dD .... re\l'acted. With a • ....u. 20 P •• lrJl:I, a ..n ... of .. clan. ot tbe !JIM!" .urt ... ot ~ t .... 1dA j\l8\ prorlMl to tba 00l"GDal 81Ilcn8 ... 8c:mU1ed. ... pleqlt. of eo\\cIa .... plaoed at. tM eoar1t1ecl arM 8Dd t.b18 ... ..t.\1Nted nth 0.15 cc ot .u.peDB1co. on. t ..... • kin .. placed to D.aN&1 pae1 cmo-l1ng the pleclelt _~. 20 ~ att..er t.lw tint, -wl1eat1GD 0.0$ 00 ot t.t. 8&p6iidCID _. added, IDd tb18 _ ... ___ ...rr 20 ldDutee em tift OQ.81~. 111 otbeZ' warda. tt. ~cm _rSlIrG _ _ with u. pem. .hr 2 bowIt. total ... t. of • ...,..t.. ..s ....

Jll .aUIId at. \De at rn hal.

D. ,A&dn1atn.t.1ca .t ~"'ey1·1

1. Pr.para\1cD at ~lai1.1

1 "t'ial. fit PD .. cU.olftd 1D 60 co at d1dUl..t .tao. 2. jppl1aat.1_ ot pl"apllrl.a:dB I

~lT lS oa at the pftp~ ... appl.1ed tor 2 1dJa--4t Pro I!ppl.led .. doatar.

,. paUent 1. .. tbiw P'01P ... t.reated witb .. ~ towr boUl"II late aU pledatt. WN raond.

a. 'or tiM of appl1oat.icm _ 1Dd1"f'1dua1. CU'd8 at paumta.

OROUP II

(lie)

.1. Source of spirochaete8' Nichola strain, rabbit inoculated em April 19, 1948.

B. Preparatica ot BIIul.IIiem.

1. Testio1es re.oved at 6.00 .1.)(. on IIIP;y 9, 19h8. 2. To the teaticJ..s, after a1Dc1Dg tharo\l8hl1' with scissora, .... added 4.0 00

of floesh UDinact1vated Rabbit 8e~ (88M rabbit), and 4.0 cc of .tarila Darmal saline. The material wu a1ud to allow tJ1a apirochaat.e8 to &Mrp trCID the tissues and beccae haaogeno,..

3. 'l'hf SUipeD8iOll'R8 oentr1tupd at a very low speed, rheostat (10. 1 S.B. Oentr1.tup) Htt1DSlo. 10 tor ; lI1Dutee.

4. In :t1_ of Spirocbutea1

20 t1eld8 wre cOUDtad, &nIrage COlmt, 2.9 per high ponr tield. The faator of the aiarosap be1ng 2.74 x J.Ob. the nuaber ot spiroch&et.ea in 1 oc .... 7.9; x J.OOJ Daber of 8piroohaetea in 0.1 oc ... 7.9; % loS.

c. heparatim and IDoculatiem ot Pat,18nta.

Por each patient tile 1'ore8Jd.D .... retracted. With a sterile 20 gaup DMdle, a aall &N& of the donIua of the inner .urtaae ot tbe toresld.n j,.tvprorl·.] to the corOD&l .ulCUII, wu acar1t1ad. .1 pledpt 01' ootton ... plaaect at the aoarU1ed area and 't.hU 11M •• 'urated with 0,1; oc ot the 8\WP8D1tian. · The to.realdn ... placed to Donal ~itiCll concealing the pledpt ent.1re:b' •. 0.05· co ot tM 8u.paaiaa .... adc»d em ; occuiODll. In other 1IOrde, tbI .uapeuica ... 1D ocataat with the pen:U tor 2 houra. Total aaouot ot the BlI8PGIiOll 0.1&0 oc. All pled .. ta d at t1M of r.onl.

D. For t.1ae ot 1IPP1l.aato1Q1l .ee 1Dd1v1dual carda ot

326. 32S • .328 • .323 • .322 • .324. 31 C(

II!: ALlEHAOOS Group II.

GROUP m. lien

A. Sowce of spiroohaeteB' Nichols .train, rabbit inoaulated on April 19,~48.

B. Preparation at EIIulaionl

1. Testioles reacwed at 6.00 A.K. on Kq 9, 1948. 2. To the testiclell, after lI1nc1ng thorougb17 with sc1allora, wu added 4.0 ac

of .fresh lD1aact1vated Rabbit 8eZ"la (a_ rabbit), and 4.0 cc of aterile normal aal.1Da. 'l'be material was aiDd to allow the spiroahaete. to _rp trClll the tUSUl!!lS and become hcaogenoua.

J. 'l'be su.peneion 11'&8 oentrituged at a T8r:! low speed, rheostat (No.1 S.B. Centrifuge) Betting No. 10 tor $ lII1nutell.

4. EnllD8ration ot Spiroahaetea.

20 t1elds 1P8re comted, averaae cotat, _ ~t9 per high power field. The tactor of tbe lIic:rOlla')pe being 2.74 x lDO, the nwmer ot apiroobutes in 1 co 11'&8 7.95 x 106, D1IIber ot epiroabaetea in 0.1 cc 1fU 7.9$ x 10$.

C. Preparation and Inooulation ot Patients.

For each patient the tona1d.n wu revacted. With a sterile 20. gauge needle, 'a aa1l area of the don_ of tbll inner awtaa. ot the toresld.n juet prox1·" to tbe coronal au1c'UB, wu soar1t1ed. .A pled",t of cottaJ. wu pl.&oed at tbe 8car1t1ed ..... ad W. 11M laturated with 0.1$ aa at the Buape:naicm. The

torellkin wu plaoed \0 :naral pae1Uca ocmceallDg tluf pledget ent1r~. 0.0'5. 00 ot the 8uape1181cm .... .&lid _ ooo .. iallll. In other worda, the suapeD8ion ... in contact with the p.m. te tID,. Total _omt of the auapenalon o.bO ao. All pledge'te aoaked at tiM of r..crval.

D. AdIId.nUtrat1on ot proph;y1.axl.81

Ute.l" 2 hours apoaUN \0 tM ,apiroohaetea, patient. in this poo", _re treated as tollon.

1.. !!eJ?!ratiOD ot Prop&lactio (CZ7Btall1ne pee Q aod1. aalt, lot No. 47121205)

To a v1&l oCIDta1n1ng SOO,OOO mt& ot peD1oil.JJJl W88 added 1$ 00 of diatdJ.led water and. 't.hU solution W8.11 Uled per patient tor 2 Jlinutes. Pro applied 'bf doctor.

C. lor tiM at ~pl1"~t1CD aee individual car~ ot patient.

GROUP IV (leD)

.1. Source of .piroobute,u Jlichola strain, rabbi t inoaulated on April 19, 1948.

B. PreparaUCIl at EIlulIiCllI

1. Testicles re.cwed at 6.00 .1.11. CIl Ka;y 9, 1948. 2. To tJ:. testicles, an.r miDcing tborougb.:q with scissors, 1188 added 4.0 co

of fresh 1m1naativated RAbbit SerlB (S8118 rabbit), and 4.0 cc ot .t.erila normal. aaJ.1ne. n. uter1al ... 88 Jlixad to allow the epirochutes to p floca the Uasue. and becCDa tu.ogenoUl.

3. TbI sUlpenaiCll" centrifuged at a ftr.Y law speed, rheostat (Ha. 1 S.B. Centritup) .. tUng No. 10 tor S llinute ••

4. lnw.ratiCll ot Spirochaetes.

20 tielda wre cOlnted, anrap aamt, _ ?t9 per bigh ponr field. The. tactor ot tbe morosagpe be1q 2.14 x ]DO, tb8 n_ber of spirocbae1ie. . in 1 00 .... 1.9, x lQOt n __ at .p1l"oohMte. 1n 0.1 co wu 1.9, x loS.

O. Preparation aDd InoculatiCll of PatieDtsl

FQL" each patient tbl tore.kill ..... retracted. With a ster1l.e 20 laup needle, a aaJ.l area ot tbe dGl'lla ot the 1rmer .urface of t.b8 tarelld.n jUlt, prcnd·.] to tbe CQL"aaal. .ulaus, ...... oar1t1ed. .1 pledget ot cottca ..... plaoed at the .carified area aDd tb18 wu lIaturated with O.lS cc ot the suapeu1oa. The f'orea1d.n .. placed to DOrul poIition aonceaUnl the pledpt _tire~. O.oS cc ot the lI-.peuiGll wu adct.d (11 2 occu1cma. ID other 1I"01"dI, the 'UlP8Dllica ..... in a(l1tact. rith the pem tar aae and a halt DoUl"l. TO'\al aM.t of +.M 8U1P8Uica O.2S cc. All pledgete soalatd at, t1lle ot re.ot'al.

D. For time of application •• e ind:I:r.Ldual aard8 of patients.

OBOUPV (1IiD)

.... Souzooe or Spirochaeteel N1chols etrain, rabb1t iDoculated AprU 19, 1948.

B. PNparat1ca or JauJ..1c:a1

l. Testicles l"DOV'ed at 6.100 A.M. Kq 9, ~48. 2. To the teet.1cle., after IWlciDg t.boroueblT with so1 .. 01"II .... adcWd 4.0 co

of freeh UD1nact1Tated Rabb1t Ser1a ( .... ~abb1\), and 4.0 oc or .terile normal. .al.1ne. 1'be .. ter1al ... m.d to all_ 'Ule apirochMt.. to ~ .. trOll tbe t18su.e Ed becOM hmaogeno'Ul.

3. n. .UlptII8101l wu C8ntri.tu8ad at a varT law IPMd,r __ ta\ (Ho. 1 a.B. OII1tr1tup) .. \tinS Ho. lO for, ainute ••

4. Bn .. raticm or Spirocbaete.·,

20 fUlda 1Mre cO\Dlted, aTarap oost ~!i. per hiab powv f1eld. The ractar of the mar_otpe being 2. Tli x JJ,IJ, the n_her of apiroohMte. in 1 cc was 7.9, x 1001 n~ of ep1roobaetee ill 0.1 cc ... 7.9, x 10'.

c. Preparat.ica and Inooulat.1ca of Pat:l.ezrt.I

0.1 00 of the • ar T .9S x 109 lPiroohMtee,... introduoe.d into tbe leftroreara.

D. !1aa of IDjeot1C1l1

SEE nffilVIDUJ.L CAJtm OF p.lmNTa.

17,. 176. l77. l78. l79. 2lO. 220. 209. 213.

~. lei Oroq> I Date. IIq 9. 1948

( )

A. SoUZOC8 at 8P1rocbea'tM I I1cbolB .torain, rabbi\ inoculated aD Jjp.rU 19. 19b8.

B. Preparat100 at Eaulalcml

1. elM ed a" 6100 A.M. GIl __ 9. 1948. 2. To the te8t1~. attar Idncd.n tharo~ nth ec1aa .... add8d 4.0 oc at

freah ,m1nact.ivaiied Babb1' ( .... nbld ) 4.0 oc ot 81ier1le DOZS1 aal1ne. t.er1al .... to allow tba spiJoocbM_ to eaarge trC%:J. tbe ~

J. t'be icm CMJrt,ri..t at a W17 low speed, rbeoetat (BG. 1 S.B. o.nt·~) -~t.1nc 10 for S l'I1nutM.

4. In at.! ot Bptrocha.frt.ea I

20 tielda ware comted.. arer.... c9Zt, 2.9 per high power field. 'fba tactar' at the lIl.SJZ008cgpe ~, 2.74 z lIP. n.tHu- ot ~ 1n 0.1 oc tfU

7.9'5 z 10=' •

. C. Preparatica. ud IDoculaU<m o! Pa t.., lJ pat.1enw --.]IIJ.cI. n. tim t was 1Doeulated at. 6:4Jj • tba last -' a 1511.1'" fbI.thad .. d 1IB8 88 tou-.. Tba lan 1DV ana .. eUB1I8ed with alcohol, .. h1ob .. w1ped. att witb cotton, CIDB drop of the 8U8p0D8~0l1 as plaoed an the l11'li (1 ... 0.022 co or 8\181*l81ca IIPPl'GX1atell' l.S9 x 105 8P1rocbMteII). Jlult.1p18 pn.ure _t.hOd W&II ,.ad t- tor ll...pcu; YBCC1Dati.ca). n. 8\J8P8D81aa r_dned 111 ccmtact with tbB .k1n tor S llinut.ee.

D. tiM or Inject.ian

SE IlIDIJ or PAT • I. I cWfn1at.raU.. at ~.

Two hours atter ....,c:1Dat1~ each paUeDt 111 tb18 1l"O"P wu giWll cral PEia1lUn.J (.300 .. 000 u:d. , esc tablew Buttered IWIScll"n - CrystRl.l.iD3 IWdcUl1u a Potud-. Lo\.o.~~).

ASn.o IE AL1ENADOS I - )(ujsru

A. SoUE'C8 or IIP~I I1cbob1 atn1D, rabbit 1Docalated GIl.lpr1l. 19, 1948.

Be ~ of Bllu lll1m1

a:1 __ 9, 1948. 2. 1'0 tbe tHt..1 cl ... atter-a:uJ.a:uI with acUsCZ"8. 1IU tlddJd 4.0 cc at

treeh wdDacUftted Babtd.. a.r. ( r t). 4.0 oc ol,aw1le aar...:L .... 'ne. n. 1&1. 1JU a1ad to a.1l.GIF ~tea to 8Ml"88 trail tbe t.1.Ruea -

3. 'lbs at a ftl7 1.0. 8ptIed, rheostat ( ~. 1. 5.13. V1t 10 tor SaiD.

c. PNparaticm IDoculaUcm 01 Pat.1aDt..

8 paUeDta 1fVe u..l. The tirS\ paliliIAIU

<IDe at ::!);~,.... o.lcc or tbe au.p-'1aD SDt.roduced ~ the let~ tonam.

D.tDeot~.

PA'l'IEftS

I. JdII1n:1atraUCD o£ Prap~.

!fWIO how-. an. ~ ..... JJM.'e\ :rec:.1ftd SJd,raYeDGUII~ hO -e. ~ llafaN1da LIlt. Jr.. 60SSlB ,....~ " Co. 1xp1rat.1CID date. Dot. ), 19Ja8 •

...

IJ,_ lJX8 n. .... , ... m

(1ltJJ!R!S)

Ae 80111'08 of ~~. JI1cbo18 at.r&1n. I'Sbb1t 1Doculat.d CII .&prU 19.

PnparaUml of !Au1.B1mI.

1. !eet.1clN ~ at 6.00 Ad. CIIl J(q 9. ~168. 2. 'fa tbe tMt.1cl1J81t aner ""nain tborcnasJ:ilT with sc18ora .... addad 4.0 oc of

frMh minacU.nt.ec1 Blhb1t Ser1B < __ rabbit). and .4.0 cc or .ter11e IaWIl _"M. 1'be _t.r1al. ..., II1Dd to all.cJIr tba 1ip1rocbMtie8 to ge trca b """----- aad be~ bewl"""".

J. DIe .capeN'm .. oaDtr1.tupcl at & 'ft'rT law speed .. rbeona\ (Ro. 1 S.B. C8DtrUup) _tt.1Dg 10 for , ldDu_.

Ja. 1a-.rat.iClO at ~ochaatM I

20 fields oo-.nteli. ~ ~t. 2.9 per hlp power t1e~ fbe ractt.r of tot. II1;roIIcope baing 2.14 x lfP. DWIbR or 1I'p1rocbMt. 1n 0.1 oc .. 7.9S z: 1£)5"

De Pnparat1ca aDd Iaaculatr1Cl1l of Pat.1.eDt.IJ 1

8 paUata 1Jftd.!tw t1rst pau.nt was 1Docrolated .\ 9106 .&..M. t.be laat. aaa ,. ...... O.l. GO os: t.a. 1aD .. T.9S x l.()5 1I'p1rocbMt.e. .. 1nt.roduoed 1Dto t.be left f arallD'lllA

D. n.. at ID,2ecUc;a.

SBE llIDIVIDUAL c.&Rm 01 PJ,fIIor._ClLa.

M-'n1at.rat.1m at ~I \\ ,Co ... ~dIrp atter jnject:S.an. each .-."" ....... 1:, 1D th1a gl"Otp ind 1 cc (300 11.) of cr,yst.e1J1M i8" c1 '''n Q i in Oil esc Lot 0. 47l1283]"&

IE~ luJanl - Cl:r\4)O m

B. PrepanLUcm at lIIulaiCiIi

1. feeUC'. ~ at, 6.00 CID 2. To tbe t.t1cl_.' .n.r atna'DI tbl:a'01l1b:b with adHanI .... addIId 4.0 oc of

boah lDlDactlya.d JIIbblt Iwwa ( ... nbId.'). aDd 1&.0 OC OIl .tariJa uan.l sal 1ne. _tar1al .. II1.JDCl to al.l.w the ~. to ~p tl"Ca tba

U88 and 1..- _""IDD'I~ 3. !ba . , lcm .. oeut.r1t at a varr l.c. apeed, I"beoatat (10. 1 s.B.

Oen ) .. t tor 6 II1D • 4. Bnwm-at1on ~ 8p tea I

1 pat,Wlt .. ..s. at. f A.JI. 0.1. 00 81_ ... 11_ G- 7.9S x lOS lIPb"oabMtIea WIlD 1Djecte into tt. lett iarsa ..

ASILO IE AIJEIW)()8 QrupoIV

(Ilujerea)