407, \," OBSERVATIONS ON THE FORMATION '" OF "BUDS" BY,THE SPIROCHMTE OF RELAPSING FEVER IN; THE LOUSE.

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1 \. 407, \," OBSERVATIONS ON THE FORMATION '" OF "BUDS" BY,THE SPIROCHMTE OF RELAPSING FEVER IN; THE LOUSE. :By LIEUTENANT-COLONEL J_' C. KENNEDY, Central Labo'ratory, BaghiJ,a& 'l'there is still much to', be done towards \ the' elucidation of the life history of Sp:irocluete recurrent'is in the body of tli~ louse. The following would appe~r to be p'~oved and generaliy accepted :-:-, ' ' (1),The spir,ochretes after ingestion by the louse very soon disappear, ' from the gut and cannot be found on dissection. ~. " i,(2) For the first four,days, th~reafter the spirochretes reniain invisible.and th~ louse is not infective.-.,,,,' '. (3) On the fifth to the sixth days the spiroyhretes are,stili invisibie but the louse is infective.. "'" I', (4) On, the seventh to th~ eighth days spirochretes appear as slender;.- forms.in the louse which is still infecti ye. ' ". ' (5) From the.tenth day o'nwards,adult:spiro~hretes appear in the lous~ which,ibn~w practically non-infecti~e. " (6) The adult (non-virulent). sprrochretes usually disappear, from the body of the, louse by the nineteenth day"but may be found up to twentyfive days..'" (7) There' is hereditary transmission of the virus in' the louse as the -offspring are infective.,:, Two stages, therefore, in the life history of the spirochreteinthe louse.are unknown~first, the stage qf invisibiljty during the first four days and,,.se~ondly, the transmis~ion to the offspring.', These 'gaps in our knowledge will nd doubt be filled when we have acquired the \ ineans of recognizing: and studying the granuie phase of spirocllretesin its entirety. Meanwhile I am induced to make an ()bserva~.tion which so far as I know has n'ot yet been recorded, 'and which in many respects is very similar to that made by Leishman in the case of S. I!utfoni in the tick, and because,there would 'appear to be some who,,do not believe that spirochretes undergo any developmental phase in ' t~e body of the louse. ' ' These' observations were' made on the con~nts of two lice which were.dissected twenty-fo~r and thi~ty,hours, respectively, after being,ren.;t0ved " from the, person of a relapsing fever case,who had been ill for about three.days. Both! lice harboured'large numbers of spirochre'tes, an cia from the size and number of the spirochretes had' pfobablybecome infected more than ten 'days previously., \ ' \ There were no spirocbretes inside t~e gut but enolmousnumbers in the, crelomic fluidanc;i round, about 'the posterior end' of the, gut. The majority were large, collected Ill, c,iumps and mptile but delicate and I o, \

2 ""',.'!, "Buds'" -by the~ Sl)iroch~te of Relapsing Fever...L ),.

3 . i Sa. // lla, \ 9a..IOa:' 12a. 13a. 'I I, \ 25a. ~ ""';'.. '.. I.. ' 2.7,. i 26a. 28.

4 If.\ /., \1. ',I \ '\" 410 " Buds" by the'spiro(fhcete of Relapsing Fever...,.". : /- granul~r In appearance. There, were also a mi~ber' 6fsmall forms. in.one case, particularly at the ante~ibrend ~f 'the body;cavity; but none wer'e found)n tlfe salivary gla~ds.. The~r presence, was ascertained by rapid ;e:lmminationof the, wet specimens, which were then fixed in osmic acid or alcohol and r,tained by Leishman, Giems~ or Heid~mhain's,stains. The presence of ~"buds" was realized.when examining ~'a slide, by darkgroijnd ill~mination, and, on" closer exan'lination th~y~ were!.fouild In several others.. Af~er a careful examination by 'both'methods~ of:.illumination;.tbere. is, no.doubt in my mind :ofthespirocbffital origin of. these "J5uds."..'... 'The.origina.lsof the accompanying drawings Wer.e made by" means \of aocam~ia lucida, magnifying about I,pOO, diameters, and many of. the. obje,cts are depicted as.they appear both 'with dark ground and ~ith.direct illu'mination.. Those figures with a following the number are from - the same objects as those b'earing tlie corresponding number~. The " buds "vary ii). sizefrolil.2 2 ft to 4'5 ft, and their situation in relation to ~he, spirochffites may 'be 'termij,la1i subterminal or lateral..they (~tain feebly with Leis}lman or Giemsa stains, but Heidenhain's brings them up better.and differentiates one or more deeper staining portions (fig~. 12a to,. 15a, 27).. 'They are,not very refractile (dry' film), but.f requently show' a., refractile granule (figs. 9, 10,. i2, 13). Bodies similar in. appearance were, fgurid lying free, presumably detached, from tlie spirochffite, and one was found in, the preparation made from an ov,um, but this may have' bebn accidental' (fig~' 28). Large'numbers of refractile granules were found in theccelo~ic fluid, in, the tissues, round ' l the gut, and some clumps in the ovar~es and ova.. It is, of' course, impossible to say which were of spirochffital origin.. \. In one of the f!peciinens from the ccelomic fluid,.it"was noted that the spirochffites had, an affinity for 'certain cellular elepjents. These cells were round and cyst~likei 4'8 ft to 6ft in size, with. a well defined margin, ari.d ;taining a faint slaty-blue,with ileishinan. A number\ showed' one, two or more chrorhatin granules at the margin (fig. 25a). These graimles un.do\j.btedly were spirochffitalarid were refractile by dark ground ilhimi-, nation (figs. 16 to, 26).. With Heidenhain's' iron hffimatoxylin they stain uniformly a deep black. Similar cells are to be'foimd' in non-infected lice but without the granules.....'. As the relapsing fever seas,?n i~ now practically over, I donot antiqipate. having much opp?rtunityfor further investigation at pr~sent.,.. DESCRIPTION OF, DRAWINGS. Figs. 1 to 7.-Appearance of spiro~hretes with gran~les and" buds" seen with dark ground. Figs. 8 to 15.-Ditto.. Compare these with figs.. 8a to 15a, wldch.are the same spirochretes seen,, with direct illumination. Stained Heidenhain's iron hrematoxylin.. Figs. 16 'to 26"Cellular 'element~ with refractile granules, with and without spirochretes i!. ', attached... Dark ground..! :',, '. Figs. 25a and 26a.-The same as figs. 25 and 26.. Leishman's stain., 0. Fig ,.." Bud "~howi~g differentiation of a deeper.staining portion.?ompare" buds" 12a, I. 13a, 14a, and.15a....,...fig. 28.'-?" bud" from, crushed ovum. ' ~.. 1 '\ '

5 J. O. Ke1'?nedy 411 \ S~~h reference~ as ~re available to me at present are for the most part the extracts publis~ed in the "Tropical -Qiseases Bulletin." " ' " NrcOLLE AND BLANC. C. B., Acad. Sc., Idem. Arch. Inst. Past. Tunis,191t.,,' FANTHAM. Ann. Trop~ Med. and PM'., Ibid.. Brit. Med. Journ., March 18, TOEPFER,. Munch. Med. Woch., 1916.,MACFIE and YORKE. Ann.,Trop. Med. and Par.,:1917. KOCH. Deut. Med. c-woch., ' PLOTZ; Journ. Exper. Med., NUTTALL. "Parasitology," 1917., MAYER.' Deut. Me'd.Woch., 1917; LEISHl\[AN. A1in. Inst. Past., TOEPFER. Deut.Med. Wo~h., 1918., ' 28 I,' I i 1\