野口英世著

Journal of Experimental Medicine

所収論文

こ の PDF は Journal of Experimental Medicine に 掲 載 さ れ た 論 文 を Rockefeller
University Press の許可（2020 年 3 月 18 日付）を得てアップロードしています。

E T I O L O G Y OF O R O Y A FEVER.
V. THE E X P E R ~ N T A L TRANSMISSION OF BARTOI~LI~ BACIL-

LIFOP,~S BY TICKS (DER~ACENTOR AI~VERSOm).
BY HIDEYO NOGUCHI, M.D.

(From the Laboratories of The Rockefeller Institute for Medical Researck.)(Receivedfor publication, June 21, 1926.)
The geographical distribution of Oroya fever, which occurs withincertain limited areas of Peru, has suggested to many students of
disease that infection is carried by some biting insect or arthropod,and Castellani and Chalmers t note particularly the analogous localization of the tick-borne disease, Rocky Mountain spotted fever.
Entomological investigation in relation to Oroya fever has not so
far implicated any particular insect, with the possible exception of
Phlebotomus verrucarum Townsend, 2 and experimental work has beenhampered by the lack of animals susceptible to infection with BartoneUa bacilliformis and methods of cultivating the organism. Theseobstacles have recently been overcome,s,* however, and the presenceof Bartonella bacilliformis in insects can now be definitely determinedboth by cultural methods and by infection experiments. There isconsiderable evidence,3-s moreover, that verruga peruviana is also
caused by Bartonella bacilliformis, as had long been suspected becauseof the similar geographical distribution of the two conditions andtheir frequent association in the same individuals.
It was not possible at the moment to undertake an investigationof insects from the infected districts, but an indirect method of approach to the problem suggested itself, that of determining whethert Castellani, A., and Chalmers, A. J., A manual of tropical medicine, London,
3rd edition, 1919.
2Townsend, C. H. T., J. Am. Med. Assn., 1913, lxi, 1717.
*Noguchi, H., and Battistini, T. S., J. Exp. Med., 1926, xliii, 851.
*Noguchi, H., J. Exp. Meal., 1926, xUv, 697.
5 Noguchi, H., and Hercelles, 0., Science, 1926, lxiv, 121.
729

730

ETIOLOGY OF OROYA FEVER.

Vsome well known carrier of infectious agents--such, for example, as
the tick which transmits Rocky Mountain spotted fever,~ tularemia, 7
and a filterable virus pathogenic for guinea pigsS--could also act asa vector of 13artonella bacilliformis under experimental conditions.
Success in transmitting the parasite under such circumstances, while
not solving the problem of the actual vector, would neverthelessfurnish a starting point for the investigation of the question.
For the specimens of ticks (Dermacentor andersoni) used in theseexperiments, I am indebted to Dr. R. R. Parker, of the United States
Public Health Service, Hamilton, Montana. The ticks were firstallowed to feed on normal guinea pigs to determine whether or notthey were free from the spotted fever virus. If the guinea pigs escaped infection, the ticks were then placed on the shaved abdominal
skin of monkeys (Macacus rhesus) in various stages of infection with
Bartonella bacilliformis and allowed to feed for periods varying from
24 hours to 6 days. Tests of infectiveness were made by feeding theticks on normal rhesus monkeys either immediately following theirremoval from the infected animal or after an interval of 12 to 14 days.
Two series of experiments were made. In the first, the periods oftick feeding were relatively short--24 hours to 4 days, and several
days were allowed to elapse between the removal of the ticks from theinfected animals and the test feedings. Infection not being obtained
under these conditions, in the second series the period of infective
feeding was lengthened to 5 days, the ticks were transferred to normal animals immediately after their removal from the infected ones,and 6 days were allowed for the test feeding. Under these circumstances the ticks proved infective.

Experiment I (Negative).
Feb. 1, 1926. Thirty ticks were placed on three infected monkeys(Nos. 7, 12, and 14), ten on each animal, and allowed to remain
attached for 1 to 4 days.
n Ricketts, H. T., J. Infect. Dis., 1907, iv, 141.
Francis, E., J. Am. Med. Assn., 1925, lxxxiv, 1243.
8 Noguchi, H., J. Exp. Med., 1926, xliv, 1.

HIDEYO NOGUCHI

731

M. rl~sus 7' had been inoculated intravenously and intradermally on Dec.
21, 1925 (41 days previously), with blood and suspensions of nodular tissue fromMonkeys 2, 3, and 4. Bartenella bacilliformis was present in the blood 4 days
after injection and on several occasions during the following month. The local
lesions were well developed by Jan. 19 and were excised* on that date. From Jan.
27 to Feb. 1 fever had been continuous, fluctuating between 104° and 105°F.
The ticks were left attached for 24 hours (Feb. 1 to 2). Fever continued until
Feb. 9. The blood had yielded cultures of Bartonella bacilliformis previously indilutions as high as 1:10,000,000, but it developed subsequently that the titerhad fallen to 1 : 10 at the time of the tick feeding.
M. rhesus 12 had been inoculated intravenously and intradermally on Jan.
19, 1926, with first generation cultures, derived from the blood of Monkeys 4, 6,and 7, and grown for 14 days on leptospira medium. The blood was positiveby culture on Jan. 25 and on Feb. 3 (6 and 15 daysafter inoculation). Theintradermal injections gave rise to small nodules, which, however, never progressed
to typical lesions but gradually receded within a month to mere traces of induration. The temperature was continuously high (104°) from Jan. 30 to Feb. 5.
The ticks were allowed to feed 72 hours (Feb. 1 to 4).
M. rhesus 14~ had been inoculated on Jan. 29, 1926, with a suspension of
nodular tissue from M. rhesus 3 and with cultures of Bartonella bacilliformis
derived from human and monkey blood, the material having been introduced
intradermally, subcutaneously, and by scarification. The temperature rose
after 24 hours, and the animal remained febrile (104-104.6°F.) for 4 days. The
ticks were allowed to feed on the animal Feb. 1 and 2. Blood taken on Feb. 8yielded cultures of Bartonella bacilliformis in a dilution of 1:10 only. The localand general symptoms progressed steadily until on Feb. 23 the blood yielded
cultures of Bartonella bacilliformis in a dilution of 1:10,000, the nodules on theeyebrows were very large and protruding, and those on the abdomen measured
1.5 × 3 cm. It is evident that the ticks were placed on the animal too early inthe course of the infection.
F e b . 16, 1926. T h r e e ticks of e a c h lot of ten were placed on thes h a v e d a b d o m i n a l skin of M . rhesus 19, a n d t w o of each lot were dissected a n d suspensions of the viscera i n o c u l a t e d i n t r a d e r m a U y i n t ot h e s h a v e d skin of t h e e y e b r o w s a n d a b d o m e n of M . rhesus 20. T h esuspensions were tested also b y culture. N o infection could bed e m o n s t r a t e d in the ticks, a n d the cultures were n e g a t i v e for ~artonella bacilliformis.

M. rhesus 19. Nine ticks, of which three had been fed on M. rhesus 7, threeon M. rhesus 12, and three on M. rhesus 14, were placed on the shaved skin of the* All operations were performed under ether anesthesia.

732

ETIOLOGY

OF O R O Y AFEVER.

V

abdomen on Feb. 16, 1925, and allowed to remain 4 days. All became attached.
The animal showed no signs of infection--no enlargement of the lymph nodes,
fever, or skin lesions. Because of prolapse of the rectum, the animal was killedby etherization on Mar. 6, 18 days from the time of tick feeding. Nothingabnormal was found at autopsy, and cultures of heart blood, bone marrow,spleen, lymph nodes, liver, kidneys, testis, and lungs were negative for Bartonella

bacilliformis.
M. rhesus 20. Inoculated Feb. 16, 1926, intradermally into the shaved skinof eyebrows and abdomen with the mixed suspensions of the viscera of ticks fed
on Monkeys 7, 12, and 14. There were small indurations at the sites of intradermal inoculation on the eyebrows after 10 days, but they disappeared within
3 weeks. The abdominal skin showed no reaction to the injections. The temperature rose to 104° or higher (104.6°) on several occasions (Feb. 19, 21, and24, Mar. 7 and 11). No swelling of the lymph nodes was noted during 42 daysof observation.

Experiment 2 (Positive).
F o u r ticks were allowed to feed on each of two infected m o n k e y s(M. rhesus 18 and M . rhesus 23) for 5 days, Mar. 24 to 29, 1926.
M. rhesus 18' had been inoculated on Feb. 15, 1926, intradermally and byscarification on the shaved eyebrow with a suspension of nodular tissue fromM. rhesus 5. On Mar. 5 and 18 the blood yielded cultures of Bartonella bacilliformis in a dilution of 1:100,000. The nodules on the eyebrows became extremely large. During the time of tick feeding (Mar. 24 to 29) the temperaturefluctuated daily between 104° and 105°. On Mar. 27 the blood was positive ina dilution of 1:10.
M. rhesus 23 had been inoculated Mar. 9, 1926, intravenously and by scarification with fourth generation cultures derived from M. rhesus 7, and intradermallywith a suspension of the nodule excised from M. rhesus 18 on that date. The
local inoculations induced in time very striking nodules and indurations, andblood taken on Mar. 22 yielded growth of Bartonella bacilliformis in a dilutionof 1:100,000. The temperature was 104°F. on Mar. 24, when the ticks wereplaced on the abdominal skin. All were moderately engorged when removed on
Mar. 29. The course of infection in this animal subsequently became very
severe both locally and systemically. There was a prolonged period of fever,and 2 weeks later spontaneous millary eruptions appeared on the face. Theanimal was sacrificed May 1, 42 days after inoculation.
On removal from the infected animals, each lot of ticks was immediately placed on a normal rhesus m o n k e y and allowed to feed fora period of 6 days. After the feeding the ticks were dissected, andmixed suspensions of the viscera were inoculated into another normal

733

RIDEYO NOGUCmmonkey. Definite, though mild, infection was induced in all theanimals, and Bartondla bacilliformis was recovered in culture fromthe blood and lymph nodes. The suspensions of the tick visceraalso yielded cultures of the organism.

~D~y~fe~p.

~46i.~ ~~6t0 12 14 I~ ~~Izla. ~"rm~,:xP,,J + + : :::++++++r~++++++++++;1
100 ++t!++t[+-~++++~++~+++~+++t; : i ~ + i ! : i i i i i i ! i ~ ~ i i i ~ !.,,.~J~
104~~r ~! ~

l0li ,t 0 ~ ] N N ; ++: + F+++ i +i N i:]+:+i ~ i i i i++:+i i i ! i++! IP!

TE:Cr-FIG. 1. M . rhesus 26.
H,~t~ A p e1Mtea0~.yofe~p.

ZI4,~6,5 T 9 11 1,3 15 ff 19 2t ~ 2~
8 10 12 14 16 tB Z0 It 24 16 ~,Tct~Ei~ ~i! iii i i ' i i.,.~;I + :+'+ + + + + + $ +.+ + + + + i + + ! ~I"++I : ; ; +: + ,.; i i :.-+ + : ; +!r,i + + + +++ J +:+I,0 o I + + 7'X !+:,..LI+ A+ t + +~ + ]'~'+ +~i,°,I)~I+++++Y++++++++++++++P++++'I•' . o l t ! l ~ + + t " ~ l + +++++ +++++++++~,]

tot++++++++++++ +++++++ ++++ +I++~i+i++++++++++~+++++t+iti:~
TL~cr-FIG. 2. M . rhesus 27.
,I~r+Tm~.'P,.o~
IC4I0~,

8 10 fl 14 16 ~ 20 ~ 24 26 ~5 ~
4 6 8 I0 fl 14 16 15 ZO 21 ~ ~: i :i~ i i ! i ' i i i i i l ! i i i ] ~ i i i i i ' i lt~

iii i i i i i i i.i !.i i i i ', ~, i i + i ~, ~, ~ ~ ~, '~ ', I! i i i i i_i ~i 'J.:X.Gk.~ ~ i~.i L,i ;~i i/~ i i-~1i i ~ ~ i l l ii iX.Z~ii-i~ill"]'.~ Ii_i ~'~..~/~., " i i i i i ~ ! i ! i P,J~ ;.;x.d,~.." ~ ]"~=+,~ i : , : !'i i i i i i ! ! :: i ~ i iM! ~-[ i i J

Io2 ~g [ i i i i i ! ! ! ! ! ! + ! ! ! ! ! J ! + i,i + i !
+~+++++++++++++++++++++++++i

TEx2-FIG. 3. M . rhesus 28.
M . rhesus 26. Mar. 29, 1926. Four ticks which had been allowed to feedfor 5 days on M . rhesus 18 were placed on the shaved skin of the abdomen andleft there for 6 fllll days. On removal, two of the four were found dead, theother two were we]] engorged. The animal began to show irregular mild fever(Text-fig. 1) 11 days from the time the feeding began, and the lymph glands in734

ETIOLOGY OF OROYA FEVER.

Vthe inguinal and axillary regions on both sides were somewhat swollen. OnApr. 14 one of the inguinal lymph nodes was excised for examination. A suspension of the tissue yielded cultures of Bartonella bacilliformis. The temperature
was 104-105°F. from Apr. 21 to 25, and blood taken on Apr. 24 yielded culturesof Bartonella bacilliformis in a dilution of 1 : 10. The enlargement of the lymph
glands has steadily progressed up to the time of writing (May 30, 1925.) No
skin lesions have developed.
M. rhesus 27. Mar. 29, 1926. Four ticks which had been allowed to feed for
5 days on M. rhesus 23 were placed on the shaved skin of the abdomen andallowed to feed for 6 full days. All the ticks became attached and were moderately engorged at the time of removal. The reaction in this animal was muchthe same as that in the foregoing monkey. Cultures made with a suspensionof one of the inguinal lymph nodes excised on Apr. 14 and with blood withdrawn
on Apr. 24 were positive in dilutions of 1 : 10. The animal has shown irregularfebrile reactions (Text-fig. 2), but no skin lesions have developed up to the timeof writing.
M. rhesus 28 (Text-fig. 3). Apr. 5, 1926, inoculated intradermally on theleft eyebrow with a suspension in 1 cc. saline of two ticks fed on M. rhesus 26
and on the right eyebrow with a suspension in 2 cc. saline of four ticks fed on
M. rhesus 27. Six-tenths of a mixture of the two suspensions was injected intravenously. The course of events in this animal was very similar to that inMonkeys 26 and 27. Small nodules appeared at the sites of intradermal injection on the eyebrows about 8 days after inoculation, but they did not progress
further. The lymph node removed from the inguinal region on Apr. 14 andblood withdrawn on Apr. 24 both yielded cultures of Bartonella bacilliformis.
A few papular eruptions had appeared on May 19. The skin became yellowish
in color about a month after inoculation and has remained so up to the timeof writing.
SUM-M.ARY.
Experiments are reported in which Barlonella bacilliformis wastransmitted from infected to normal rhesus m o n k e y s b y the bite of
the tick, Dermacentor andersoni. A long period of feeding, b o t h on
the infected animal and on the normal animal subjected to infection,
was required in order to secure positive results. The infection transm i t t e d b y the ticks was mild, b u t definite, as shown b y the recoveryof Barlonella bacilliformis from the l y m p h nodes and blood.