NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
13
folds of the paper. These tubes were then placed in a small dish standing in
a basin of water. No corks are necessary if the paper is pushed well down
to the bottom as the bugs cannot climb up the glass. On taking the tubes
to the bedside, the paper containing the bugs was gently drawn up to , the
mouth of each tube with a pair of forceps and the tubes inverted over the
patient's skin, shortly after which the bugs came out and began to suck blood,
and when fully gorged returned readily to the folds. Bugs fed in this way live
and breed as in nature, and can be fed either by day or night.
On March 28th, a large number of bugs were obtained from many sources
and were placed in a jar containing some crumpled filter paper ; the next day
suitable specimens were removed with a fine pair of forceps and placed in a
tube ready for use.
Experiment I, Case 11.—On March 29th, eleven bugs were fed on this
patient, in a film of whose finger blood nine parasites had been found. On
March 30th, 31st, and April 2nd the bugs were again fed although no parasites
had been found in blood films on those dates. On April 2nd one of the bugs was
dissected and the mid-gut contents which contained fresh and altered blood were
examined in two separate films under cover slips. In one of the preparations
I s a w a p y r i f o r m - s h a p e d f l a g e l l a t e a c t i v e l y m o v i n g a b o u t a m o n g t h e c o r p u s c l e s .
Its structure was difficult to make out in the fresh condition, but a hyaline
nucleus situated about the centre of the body was visible. I was unable,
however, to make out the blepharoplast or any granules in the protoplasm.
Its movements were less active than those of the flagellates found in Anopheles
stephensi, its flagellum exhibiting slow lashing movements.
The flagellates of Anopheles stephensi are extremely active, darting across
the field of the microscope, then suddenly altering their course and moving
in the opposite direction ; their flagella exhibit vibratile rather than lashing move-
ments. The movements of this flagellate of the bug were more like those
of the flagellates of Culex fatigans and Culex pipiens. I was at first in doubt
where this flagellate came from, for, on examining the saline solution used in my
dissections, I could find no organisms of this nature. On attempting to make
a stained preparation the parasite was unfortunately lost, so I was unable to
study it further. Thinking that it might possibly prove to be the fully developed
Leishman-Donovan body, I determined to go on with my experiments with bugs.
It is not necessary to give details of the dissections of 41 bugs examined
between April 2nd and 13th, as I was unable to procure a suitable case in
hospital and all the dissections proved negative.
On April 14th I was shown by Mr. Haller a case of Kala Azar in a
European, in the last stages of the disease, suffering from very severe diarrhœa.
No blood film was examined in this case, Six bugs were removed from the
14
patient's bed, and I asked the relatives to send me any others they could find, but,
owing to the death of the patient, they were never sent.
On April 15th I dissected one of the bugs, and on staining the film
found a large mononuclear leucocyte containing three parasites (Fig, 4). From
the appearance of the mid-gut contents, it was most probable this bug had last
fed on the patient on April 13th ; the parasites had therefore been in the bug
about 40 hours. They stained well with Romanowsky's stain, their outlines
being distinct, and in every way they were similar to those found in smears made
from splenic blood. Another bug dissected on the same day showed a free
parasite (Fig. 5), which also stained well, no leucocyte being found anywhere
near it. The remaining four bugs contained no parasites.
From April 16th to May 14th I was unable to obtain a good case in
hospital. A few bugs, however, were fed on a patient on April 20th, on which
day a single parasite was found in a blood film. One of the bugs dissected four
hours after the last time it was fed, contained a parasite in a polymorphonuclear
leucocyte (Fig. 3) which was lying unchanged in the protoplasm of the cell.
On May 16th a suitable case, a boy of 16 with cancrum oris, general
œdema and severe diarrhœa, was admitted into hospital.
Experiment I, Case 12. —On May 16th a film of blood was examined and
found to contain 32 parasites, and on this day 22 bugs were fed on the patient.
They were fed again on the 17th, a blood film taken shortly before the feeding
containing 52 parasites. On May 18th, 15 parasites were found in a blood film,
and the bugs were again fed. On the 19th I dissected one of the bugs, and found
two free parasites in the film (Fig. 8). From the appearance of the cells in the
mid-gut contents, this bug probably fed last on the 18th. Both parasites
showed the earliest stages of development, their protoplasm stained faintly blue
and the macro-nuclei, though still towards the periphery, appeared granular.
In one, the macro-nucleus and protoplasm contained a single vacuole, while in
the protoplasm of the other there were three.
The bugs were fed daily from the 19th to the 31st, and on each day a blood
film was examined and found to contain parasites in varying numbers. On
June 1st I dissected an adult female bug which had last fed between 6
and 7 P.M. on the 31st, and found 15 free parasites in the film. Two of
the parasites are seen in Fig. 6, the larger showing distinct signs of
development. Its protoplasm has increased in volume, the macro-nucleus much
enlarged is granular, and though still touching the periphery is extending
towards the centre of the parasite. The micro-nucleus, somewhat thicker,
is lying on the periphery of the cell and adjacent to it a vacuole is seen. The
smaller parasite is stained feebly throughout and seems to be degenerating. What
appears to be the remains of a leucocyte is seen lying close to the parasites,
15
its nucleus is represented as a faintly stained mass, and the protoplasm is granular
and indistinct.
Fig. 7 represents another parasite from the above film at about the same
stage of development. Its macro-nucleus, more central, is much enlarged, and
the micro-nucleus, somewhat thicker, lies near it. The protoplasm, stained deep
blue, contains two large and two small vacuoles, but there are no granules in it.
No sign is seen of the commencing formation of the vacuole-like area, or any
evidence of the division of the parasite.
On June 4th another female bug was dissected, and in the slide a parasite
showing further development was found (Fig. 9). This parasite, measuring 3 . 8μ , in
its greatest breadth, is much rounder than the one described above (Fig. 6) ; its
protoplasm, stained deep blue, contained three vacuoles. The macro-nucleus,
much enlarged, is pear-shaped and lying well within the parasite, while the
micro-nucleus, which is slightly enlarged, lies about the centre of the cell. A
circular shaped body which represents the vacuole-like area, stained pink by
Romanowsky's method, is seen lying between the micro-nucleus and the periphery
of the parasite. An irregular mass, the remains of a leucocyte, is lying adjacent
to the parasite. The bug from which this parasite was recovered was fed last on
May 31st between 6 and 7 P.M., and was dissected at 7 A.M. on June 4th, the
parasite had therefore been in the mid-gut of the bug about 82 hours.
On June 4th I found it necessary to feed the remaining bugs on another case
of Kala Azar, but the bug from which the above parasite was obtained had ap-
parently not fed, as there were no fresh red cells in its mid-gut. Four bugs were
dissected on the same day with negative results. On June 5th another female
bug was dissected, and eleven parasites were found in the slide, six of these
are shown in Figs. 10, 11 and 12 . In Fig. 10, two elongated pyriform
parasites measuring 5μ and 3.5μ are seen lying in close apposition. In the
larger the macro-nucleus, much enlarged and containing a vacuole, occupies
about the whole of the posterior pole, while in the other it is smaller and lies to
one side touching the periphery ; both the micro-nuclei, somewhat thickened,
are curved, one being concave towards the macro-nucleus, the other convex.
The protoplasm of both the parasites is stained a deep reddish blue and contains
neither vacuoles nor chromatin granules. A brush-like structure consisting of
many pink staining filaments is seen protruding from the anterior end of the
larger parasite and represents the early stage of the flagellum. In the smaller
parasite it is only just protruding and appears as a blunt rod, stained pink.
Both the flagella pass out from the vacuole-like areas which are shaded in the
figure.
Fig. 11 represents a parasite from the same film showing signs of longitu
-dinal division. The macro-nuclei, circular in shape and finely granular, are
16
seen lying on each side of the central line. The micro-nuclei are lying on the
outer sides of each parasite, adjacent to the macro-nuclei. The two vacuole-
like areas are clearly seen, and protruding from them is a small rod, which stains
pink, and represents the commencing formation of the flagella. A large vacuole
lies between the two parasites, showing that separation is beginning at this point.
The protoplasm of both the cells, staining dark pink, contains no vacuoles or
granules. Each parasite measures 4. 8μ in length and 1.5μ in breadth.
Fig. 12 shows two attenuated parasites attached by their posterior poles
only. The two macro-nuclei, considerably elongated, are situated about the
centre of each cell, while the micro-nuclei are in close proximity to them. The
protoplasm of both are stained reddish and contain no granules or vacuoles.
Each cell measures 5 μ in length and .8μ in breadth. In the same film there
were two other parasites showing early signs of development and three elongated
forms without flagella.
The remaining bugs were dissected on the 5th and 6th with negative
results.
During the months of June, July and August a large number of bugs were
fed on persons suffering from Kala Azar, in whose peripheral blood only a few
parasites could be found, but the dissections of all these bugs proved negative.
In September three cases of Kala Azar suitable for feeding experiments were
admitted into hospital. The first of these, a young woman aged 22, was suffer-
ing from cancrum oris, general œdema and diarrhœa ; I made a film from her
finger blood and found 21 parasites, after which a number of bugs were fed. I
was unable, however, to continue these feeding experiments as the patient's friends
removed her from hospital. No parasites were found in the bugs.
The second case was a boy of 17 with general œdema and diarrhœa, in a
film of whose peripheral blood I found 11 parasites. A number of bugs were fed
on three successive days, blood films taken before feeding showing many para-
sites ; as the patient died suddenly I was unable to feed the bugs again and the
examination of their mid-gut contents showed no parasites.
The last case was that of a boy aged 12 who had been in hospital on a
previous occasion when I had found the parasite in his peripheral blood ; he was
now very ill suffering from uncontrollable diarrhœa. On the 10th, three days after
his return to hospital, when examining him I found in his bed a large number of
bugs in all stages of development, showing they had been in the bed for some
time ; a blood film examined on this day contained 27 parasites. The patient
who occupied the bed before this boy was admitted, had been suffering from
pleurisy. I removed some of these bugs for dissection and the rest were des-
troyed ; on the 14th an adult female bug was dissected and in the film I found
three parasites, two of which resembled the attenuated forms shown in Fig. 12.
17
They were attached from their posterior poles to about the centre of their
bodies ; the circular macro-nuclei were granular in appearance and situated about
the centre of each parasite. The micro-nuclei were lying close beside the macro-
nuclei, touching the periphery of each parasite. There were no vacuole-like areas
or flagella. The third parasite was oval in shape containing a large circular
macro-nucleus with the micro-nucleus in close apposition to it. There was no
distinct vacuole-like area to be seen, but a diffused pink stained mass was situated
at some distance from the macro-nucleus ; it was difficult to decide from these
appearances what change was taking place in the parasite.
The remaining bugs taken from the bed were dissected but no parasites
were found. On the 11th six bugs were fed on the patient for the first time
shortly before this 30 parasites having been found in a film of finger blood. The
bugs were again fed on the 12th and 13th at approximately the same hour as on
t h e 1 1 t h a n d o n t h e s e d a y s t h i r t y - t w o a n d s i x t e e n p a r a s i t e s r e s p e c t i v e l y w e r e
found in blood films. The patient died on the 13th. The bugs were fed daily
from the 13th to the 17th on a young monkey and were all dissected on the 17th
and in two of the films three parasites practically unchanged were found.
Assuming that the parasites had been ingested by the bugs on the 13th, they
had been in their mid-guts about 96 hours. I was unable to find any parasite
showing development.
In the bug I could find no large granular blue forms which are so charac-
teristic of cultures at 22°C. All the most advanced forms were considerably
smaller than those seen in cultures and in each parasite the micro-nucleus lay
close up to the macro-nucleus.
Further work on the development of the parasites can alone confirm these
observations, and it would be unwise to be too dogmatic regarding the precise
mode of their development.
The discovery of the development of the Leishman-Donovan body in the
bed bug has led to my endeavouring to ascertain the distribution of the species
of bug used in my experiments. Through the kindness of medical officers,
assistant surgeons and hospital assistants I have been able to examine a large
number of specimens of bed bugs from various parts of India, Assam and Burma,
and I have found, so far, without exception that they are identical with the Madras
bed bug. In order to compare this bug with the typical Cimex lectularius, L. I
obtained some living specimens of this species from the slums of London and
found that the Indian bed bug is Cimex macrocephalus, Fieb. Recently I
sent some specimens of Indian bed bugs to Mr. Distant who confirmed my
observation that they are Cimex macrocephalus.
It is generally believed, as is stated by Mr. Distant, that Cimex lectularius, L.
is distributed throughout India, while the presence of Cimex macrocephalus, Fieb.
D
18
has only been recorded from Burma (Bhamo). Further investigations regarding
the distribution of the two species are in progress.
CONCLUSIONS.
1 . T h e L e i s h m a n - D o n o v a n b o d y o c c u r s f r e q u e n t l y i n t h e p e r i p h e r a l
circulation in cases of Kala Azar in Madras, the parasites being seen in the
leucocytes but never in the red cells or free in the plasma
.
2 . I n c e r t a i n c a s e s w i t h e x t e n s i v e u l c e r a t i o n o f t h e l a r g e i n t e s t i n e t h e r e
occurs an increase of the polymorphonuclear leucocytes and many of these
cells then contain parasites.
3 . T h o u g h t h e L e i s h m a n - D o n o v a n b o d y h a s b e e n r e c o v e r e d f r o m t h e
mid-guts of lice, Pediculus Capin's, it was never found in Pediculus corporis,
Culex fatigans, Anopheles stephensi, Stegomyia sugens, or the tick (Ornithodoros
savignyi).
4 . T h e p a r a s i t e s c a n b e r e c o v e r e d f r o m t h e m i d - g u t s o f b u g s(
C i m e x
m a c r o c e p h a l u s ) ,
f e d o n c a s e s o f K a l a A z a r , a n d k e p t a t r o o m t e m p e r a t u r e
(80—82° F.), and these parasites have in a few cases shown considerable
development.
I take this opportunity to thank Majors Robertson, Donovan, Foulkes, and
Captain Long for their kindness, in permitting me to examine the cases of Kala
Azar under their treatment.
NOTE.
Since writing the above I have found in, the bed bug (Cimex macrocephalus,
Fieb.), all the intermediate stages of development and numerous fully developed
flagellates similar to those seen in cultures of splenic blood. These will be des-
cribed in detail in another paper.
19
REFERENCES.
1. Rogers.—Lancet, July 23rd, 1904.
2. Rogers. —On the development of flagellated organisms (Trypanosomes) from the
spleen protozoic parasites of cachexial fevers and Kala Azar.
Quarterly Journal of Microscopical Science, Vol. 48, No. 191, N. S.,
November 1904.
3. Christophers. —On a parasite found in persons suffering from enlargement of the
spleen in India (Third Report). Scientific Memoirs by officers of
the Medical and Sanitary Departments of the Government of India
(New Series), No. 15.
4. Leishman and Statham. —The development of the Leishman body in cultivation.
Journal of the Royal Army Medical Corps, Vol. IV, No. 3, March
1905.
5. Rogers.—Lancet, June 3rd, 1905, page 1484.
6. Rogers. —Proceedings of the Royal Society, Series B, Vol. 77, page 285
.
7. Manson and Low.—British Medical Journal, January 23rd, 1904, page 183.
8. Manson and Low.—British Medical Journal, July 2nd, 1904, page
1 1 .
9. Statham.—Journal of the Royal Army Medical Corps, Vol. IV, No. 3, March
1905, page 326.
10. Manson. —The Lane lectures on Tropical Diseases for 1905, page 146.
11. Manson.—British Medical Journal, September 17th, 1904, page 657.
12. Rogers.—Lancet, June 3rd, 1905, page 1486.
13. Bentley.—British Medical Journal, September 17th, 1904, page 655.
14. Ross.—The Journal of Hygiene, Vol. 6, No. 2, April 1906, page 104.
15. Novy, MacNeal, and Torrey. —Science, Vol. 23, February 9th, 1906, pages
207-208.
16. Christophers. —The Anatomy and Histology of Ticks. Scientific Memoirs by
officers of the Medical and Sanitary Departments of the Government
of India, New Series, No. 23.
17. James. —Malaria in India. Scientific Memoirs by officers of the Medical and
Sanitary Departments of the Government of India, New Series,
No. 2.
18. Distant.—Fauna of British India, Rhynchota, Vol. II, pages 410-411.
EXPLANATION OF PLATE.
Fig. 1. Large mononuclear leucocyte containing seven parasites, one of which lies
superimposed on the nucleus. Stained by Romanowsky's stain. Magni-
fication 600 diameters.
Fig. 2. Polymorphonuclear leucocyte containing four parasites. Stained by
Romanowsky's stain. Magnification 600 diameters.
Fig. 3. Polymorphonuclear leucocyte containing one parasite. Stained by
Romanowsky's stain. Magnification 600 diameters.
Fig. 4. Large mononuclear leucocyte containing three parasites, recovered from the
mid-gut of a bug taken from the bed of a case of Kala Azar. Stained by
Romanowsky's stain. Magnification 600 diameters.
Fig. 5. Free parasite recovered from mid-gut of a bug from bed of case mentioned
above. Stained by Romanowsky's stain. Magnification 1,35o diameters.
Fig. 6. Two free parasites recovered from a bug last fed on May 31st. What appears
to be the remains of a leucocyte is seen lying near them. Stained by
Roman owsky's stain. Magnification 1,35o diameters.
Fig. 7. Another free parasite from same bug as above. Stained by Romanowsky's
stain. Magnification 1,350 diameters.
Fig. 8. Two free parasites recovered from bug fed on case 12. The parasites were not
in close apposition in the film, only being shown thus for purposes of com-
parison. Stained with Romanowsky's stain, Magnification 1,350 dia-
meters.
Fig. 9. Free parasite showing distinct development, the vacuole-like area is seen as a
circular body lying between the micro-nucleus and the periphery of the
parasite. The remains of a leucocyte is seen lying near the parasite.
Stained by Romanowsky's stain. Magnification 1,350 diameters.
Fig. 10. Two elongated forms recovered from bug last fed on May 31st. Stained by
Romanowsky's stain. Magnification 1,350 diameters.
Fig. 11. Elongated parasite showing partial fission. Stained by Romanowsky's stain.
Magnification 1,350 diameters.
Fig. 12. Two attenuated forms showing almost complete separation being attached
only by their posterior poles. No vacuole-like area or flagella are seen.
Stained by Romanowsky's stain. Magnification 1,350 diameters.
Fig: 1.
Fig: 2.
Fig: 3.
Fig: 5.
Fig: 6.
Fig: 4
Fig: 7.
Fi g : 8 .
Fig: 9.
Fig: 10.
:Fig: 11.
FOR PAPER ON DEVELOPMENT OF THE LEISHMAN-DONOVAN BODY IN THE BED BUG.
Photogravure.
Survey of India Offices Calcutta July
. 1906.
No. 27.
( NEW SERIES.)
SCIENTIFIC MEMOIRS
BY
OFFICERS OF THE MEDICAL AND SANITARY DEPARTMENTS
0F THE
GOVERNMENT OF INDIA.
PRELIMINARY REPORT ON THE DEVELOPMENT OF THE
LEISHMAN-DONOVAN BODY IN THE BED BUG.
BY
CAPTAIN W. S. PATTON, M.B, I.M.S.
(On special duty.)
ISSUED UNDER THE AUTHORITY OF THE GOVERNMENT' OF INDIA
BY THE SANITARY COMMISSIONER WITH THE GOVERNMENT
OF INDIA, SIMLA.
CALCUTTA:
OFFICE OF. THE SUPERINTENDENT OF GOVERNMENT PRINTING, INDIA.
1907
Price eight annas or 9d.
ANNUAL REPORT
OF THE
CIVIL VETERINARY DEPARTMENT, BENGAL,
AND OF THE
BENGAL VETERINARY COLLEGE
FOR THE YEAR 1903-1904.
CALCUTTA:
PRINTED AT THE BENGAL SECRETARIAT PRESS.
1904.
[ Price—Indian, 8 annul; English, 9d.]
ANNUAL REPORT
OF THE
CIVIL VETERINARY DEPARTMENT, BENGAL
AND OF THE
BENGAL VETERINARY COLLEGE
FOR THE YEAR 1903-1904,
Calcutta:
PRINTED AT THE BENGAL SECRETARIAT PRESS.
1904.
Published at the BENGAL SECRETARIAT BOOK DEPÔT,
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