NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
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APPENDIX,
Methods of examination of the blood,
Any one who wishes to fully understand the life history of the malaria para-
site or who desires to he able to diagnose and treat cases of malarial fever success-
fully, must he thoroughly familiar with the appearances assumed by the para-
site in human blood. In order to acquire this knowledge it is absolutely neces-
sary to be able to make preparations of blood for examination, and to he able to
recognise the parasite in such preparations. For the sake of any student who
wishes to take up the study of malaria seriously a few remarks on the methods
of examining blood for malarial parasites may therefore be added. It should be
noted however that no directions given in a book are of any value compared to
the actual demonstration of real parasites by one who is familiar with the subject,
and no student should leave college without having made it his duty to obtain
such a demonstration from one of bis professors.
"It is impossible to make reliable examinations of the blood for
malarial parasites without first being familiar with the ordinary appearances
of normal blood and the more common pathological changes.*"
This axiom cannot be too strongly impressed upon all who enter upon the
microscopic study of malaria parasites, and many pages might be filled with the
errors that have been made by observers who have confused normal constituents
of the blood, such as blood platelets and even leucocytes, to say nothing of
such common pathological changes in the red corpuscles as vacuoles, crenations,
fissures, etc., with malaria parasites.
Before commencing to study the appearances of malarial blood it is essential,
therefore, for every one to make themselves familiar with the appearances of
normal blood, and to be able to distinguish differences in the colour, size, and
shape of the red corpuscles, and to recognise such changes in them as crenations,
vacuoles, and fissures, and also to be thoroughly familiar with the different
varieties of leucocytes and with the blood platelets.
For the examination of malaria parasites a good microscope with a one-
twelfth inch oil-immersion objective is necessary. The blood may be examined
either in the fluid state or in dried and stained preparations. The advantage of
examining fluid specimens is that the living movements of the parasite and the
changes that take place in it can be watched, but in stained preparations the
detailed structure is more readily seen. Whenever possible both methods should
be employed simultaneously and results compared. The articles required for
preparing fluid specimens of blood are :—-(1) Thin cover glasses, (2) glass slides,
(3) a pair of forceps for holding the cover glasses, (4) alcohol, (5) a surgical
needle.
The thinnest and best cover glasses and slides only should be used and they
should be absolutely clean and well polished. Immediately before being used
the slides and cover glasses should be rewashed in alcohol, thoroughly dried and
polished with a silk cloth, and placed on a sheet of white paper under a glass
* Lectures on the Malarial Fevers by W. S. Thayer, M.D., Professor of Medicine
in the Johns Hopkins University.
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to find parasites in such cases, and to any one who has had little or no practice
in the examination of blood I would say, (1) Do not commence to search for
parasites until you have found a typical case like that of chart I or chart II
(page 29) ; and (2) Do not search in any case that has been given quinine even
in small doses as this drug quickly causes the majority of the parasites to
disappear from the blood.
If typical cases like those of charts I and II are not available, the following
plan may be recommended. Go to a native village in the evening and take the
temperatures of half a dozen children with enlarged spleens. It will almost
certainly be found that at least one or two of the children have a rise of
temperature, though they may be playing about apparently quite well. In
return for a small present these children will allow their blood to be examined
as frequently as may be desired, and the finding of parasites in their blood
is generally an easy matter. Among the children of most Indian villages a
greater variety of cases of pure malarial fever will in fact be encountered than
are likely to be met with during many months' examination of hospital
cases.
For making stained preparations dried blood films must be used. A straight
surgical needle, the eye of which has been removed with a pair of forceps,
and glass slides are all that are necessary for taking these films. Cover glasses are
not required.
The finger is pricked and a clean glass slide is lowered on to the drop of
blood that exudes so that it touches the drop about half-an-inch from one end
of the slide. The slide is then held between the finger and thumb of the left
hand and the shaft of the needle (which is held by the point) is placed crosswise
on the slide over the drop of blood. (Fig. 3).
FIGURE 3.
The blood will run along the under surface of the needle which is then drawn
evenly along the slide to the end, thus making a broad thin film of blood.
Films prepared in this way should be allowed to dry in the air and then
fixed by immersion in absolute alcohol for half-an-hour.
It is better for the beginner to get accustomed to one stain than to attempt
many different methods, and the best stain for malaria parasites is undoubtedly
that recommended by Romanowsky. This is a triple stain consisting of a
combination of a particular kind of Eosin and medicinal methylene blue. It
can now be obtained in the form of compressed tabloids * and is therefore very
convenient to use. Each tabloid should be dissolved in 10 cubic centimetres of
* From Messrs. Burroughs, Wellcome & Co., London.
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pure methyl alcohol, and the stain is then ready. If this stain is used, pre-
vious fixation of the Hood films is unnecess ary. After the film has dried in
the air it is covered with a few drops of the stain. Half a minute afterwards
an equal quantity of distilled water is added to the stain, and the slide is
moved about so as to thoroughly mix the water with the stain. This mixture
is allowed to remain on the film for from ten to twenty minutes, and is then
washed off with a gentle stream of distilled water. The slide is then allowed to
dry in the air and examined. It is not necessary to cover the film with a
cover-glass ; the oil for the oil-immersion lens may be placed directly on the
film of blood.
By this method the red blood corpuscles are stained pink and the blood
platelets and the nuclei of the leucocytes a deep ruby-red.
Polynuclear leucocytes will be recognised by their large irregular nucleus.
Their body protoplasm is unstained and contains fine red granules (Plate VII,
Fig. 11).
Large mononuclear leucocytes have generally a large oval or horse-shoe
shaped nucleus and faintly blue stained body protoplasm which is free from
granules. They may contain grains of intensely black malarial pigment
(melanin) which should be carefully looked for as they are a sign of
recent infection (Plate VII, Fig. 12 )
Lymphocytes have a round, ruby-red nucleus surrounded by a thin rim
of blue stained protoplasm.
Eosinophile leucocytes have generally two pear-shaped nuclei joined together
at the top, and numerous large, round, red granules scattered through the cell,
which make them very easily recognisable.
The blood platelets are deep ruby-red, and will generally be found in groups
looking something like bunches of grapes. A few are seen in Plate VII,
Fig. 11. Many beginners mistake them for the spores of malaria parasites.
Malaria parasites are very easy to detect in the red blood corpuscles by this
method of staining. Young " ring " forms of the parasite in the red blood
corpuscles present the following characters :—
(1) A brilliant red dot at the periphery, (the nucleolus of the parasite) ;
(2) A clear space almost unstained (the nucleus of the parasite) ;
(3) A circle of blue stained protoplasm surrounding the clear nucleus
(the boly of the parasite). Such a parasite is represented in
Plate VII, Fig. 8.
Larger forms of the parasite present the same characters, viz.:—
(1) One or more red patches of chromatin ;
(2) A clear space ;
(3) A large blue stained body, in which grains of black pigment can be
plainly seen (Plate VII, Fig. 1).
The red corpuscles containing parasites may be variously altered. In
simple Tertian infections, for example, the corpuscles containing parasites
may be swollen to more than double their original size, and with this
stain they may be infiltrated with red stained dots to such an extent that the
parasite itself is almost hidden (Plate VII, Fig. 2). When corpuscles studded
with these red dots are seen, we may be sure that the case is one of an
infection with simple Tertian parasites. When the parasites have arrived at the
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segmenting stage it will be seen that the patches of chromatin have divided
up into a number of parts, each of which represents a "spore." A thin rim of
corp uscle may still be seen round the group of red stained spores, and near
the centre of the group a mass of black pigment will be collected. (Plate VII,
Figs. 3, 4, and 7.)
The sexual forms of simple Tertian and Quartan fevers are difficult to
distin guish in stained specimens from the full grown asexual forms, but the
sexual forms of malignant Tertian fever, viz.-" Crescents," will be very easily
recognised from their characteristic shape and from the grains of pigment
collected at their centre. They are stained light blue with Romanowsky's
stain, and near their centre, partly hidden by the pigment, a mass of red
stained chromatin will be seen. ( Plate VII, Figs. 10 and 11.)
Having learnt to find and recognise parasites the student will next desire
to know how to distinguish the three forms, simple Tertian, Quartan, and
malignant Tertian from each other. In the very young stage before the
parasites have acquired pigment, this is a difficult matter, but in later stages it
is comparatively easy. At a microscopical examination the points of distinc-
tion depend on the following observations :—(1) The stages of the parasite
which are found in the peripheral ( finger ) blood ; (2) The effect of the para-
sites on the red blood corpuscles containing them ; (3) The character of the
amœboid movements of the parasite ; (4) The number of spores into which
the parasite divides up ; ( 5 ) The character of the sexual forms. ( 6 ) The
character of the pigment seen in the parasite.
The differences in the three species of parasite in the above respects may be
tabulated as follow : —
Species of
parasite.
Quartan
Benign
Tertian.
Malignant
Tertian.
Stages found
in the
peripheral
(finger) blood.
All stages .
All stages
Only young
"ring" forms
and crescents.
Very rarely
an occasional
segmenting
form.
Effect of the
parasites on
the red blood
corpuscles
containing
them.
The corpuscle
is practically
unaltered.
The corpuscle
is very much
swollen and
decolourised.
The corpuscle
is generally
shrunken and
decolourised.
Never swol-
len.
Character of
the amœboid
movements of
the parasite.
Very sluggish
so that the
parasite is
generally
oval or more
or less rect-
angular.
Very active
so that the
parasite is
irregular in
shape.
Very active
and with a
great tenden-
cy to assume
the ring
form.
Number and
arrangement
of spores
8-10. Arranged
with great
r e g u la rity
" like
a
daisy."
20-23. Not so
regularly ar-
ranged.
9-10. The
s e g menting
form only oc-
cupies about
½- of the
red corpuscle
Character
of the
sexual
forms.
Round.
Rarely
seen.
Round
Cr e s cent
shaped.
Character of
the pigment.
Coarse black
grains.
Fine brown
granules or
rods.
Fine brown
granules or
rods.
PLATE I.
1. Normal red blood corpuscle. 2. Red corpuscle containing malignant
Tertian parasite. 3. Larger form of the same parasite containing pigment in
short rods. 4. 5. Segmentation of the parasite. 6. The spores have escaped
from the red cell and are free in the plasma. 7. A young sexual form (crescent)
developed from a similar parasite to 3. 8. Male sexual form. 9. Female
sexual form. 10. 11. Further development of these forms. The remains of
the red blood corpuscle still surround them. 12. Free male form which has
developed "flagella". 13. Free female form. 14. Fertilization of the female
by a flagellum. 15. Travelling vermicule. 16. 17. Zygotes in the stomach
wall of the mosquito. 18. The sporozoites fully developed have burst from the
zygote.
PLATE II.
DIAGRAM OF A FEMALE ANOPHELES.
PLATE III.
FIG II.
HEAD OF A MALE ANOPHELES.
FIG III
HEAD OF A FEMALE ANOPHELES.
FIG IV.
HEAD OF A FEMALE CULEX.
PLATE IV.
EGG OF ANOPHELES
a Front view b Side view
A FULL GROWN LARVA OF ANOPHELES CULICIFACIES
[Drawn by S. R. Christophers M. B. (Vict.)]
PLATE V.
WING OF ANOPHELES CULICIFACIES (GILES)
WING OF ANOPHELES ROSSI
PLATE VI.
WING OF ANOPHELES JAMESI (THEOBALD)
WING OF A CULEX (C. FATIGANS)
PLATE VII.
FIG. 1.
FIG. 2.
FIG. 3.
FIG. 4.
FIG. 5.
FIG. 6.
FIG. 7.
FIG. 8.
FIG. 9.
FIG. 10.
FIG. 11.
FIG. 12.
PLATE VII. Malaria parasites in the blood of man. X 1,000. Romanowsky's stain. Fig. 1.—Nearly full
grown Benign Tertian parasite. The red corpuscle containing it is swollen and decolourised; Fig. 2.—A similar
parasite almost hidden by the stippling of the cell; Figs. 3 & 4.—Segmenting stages of Benign Tertian parasite;
Fig. 5.—Sexual form of Benign Tertian parasite; Fig. 6.—Nearly full grown Quartan parasite; Fig. 7.—
Segmenting stage of Quartan parasite; Fig. 8.—Ring form of Malignant Tertian parasite; Fig. 9.—Pre-
segmenting stage of Malignant Tertian parasite; Figs. 10 & 11.—Crescents (sexual forms of Malignant Tertian
parasite). In Fig. 11, a polynuclear leucocyte and a few blood-platelets are also seen; Fig. 12.—A "large
mononuclear" leucocyte containing blocks of malarial pigment (melanin).
Microphotographs by Dr. C. Caleb, from the Author's preparations.
PLATE VIII.
FIG. 1.
FIG. 2.
FIG. 3.
FIG. 4.
FIG. 5.
FIG. 6.
PLATE VIII. Fig. 1.—Stomach of a mosquito showing "zygotes" x 60; Fig. 2.— The same x 300 showing
five zygotes; Fig. 3.—The salivary gland and duct of an Anopheles as it appears when dried and stained.
The sporozoites which were present in this preparation are not apparent in the reproduction; Fig. 4.—
Sporozoites from the salivary gland of an Anopheles, dried and stained x about 700; Figs. 5 & 6.—Other
parasites sometimes present in the blood of man; Fig. 5.—An embryo of "Filaria Sanguinis Hominis
Nocturna"; Fig. 6.— The parasite of "Surra."
Microphotographs by Dr. C. Caleb, from the Author's preparations.
ANNUAL REPORT
OF THE
BENGAL VETERINARY COLLEGE
AND OF THE
CIVIL VETERINARY DEPARTMENT, BENGAL,
FOR THE YEAR 1908-1909.
CALCUTTA:
BENGAL SECRETARIAT BOOK DEPÔT.
1909.
[Price—Indian, 11 annas; English, 18.]
ANNUAL REPORT
OF THE
BENGAL VETERINARY COLLEGE
(including the Reports of the Research Laboratory and the
Glanders and Farcy Department in Calcutta
and Neighbourhood)
FOR THE YEAR 1908-1909.
CALCUTTA:
BENGAL SECRETARIAT BOOK DEPÔT.
1909.
Published at the BENGAL SECRETARIAT BOOK DEPÔT,
Writers' Buildings, Calcutta.
OFFICIAL
AGENTS.
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TABLE OF CONTENTS.
General Administration . . . . . . . . . .
Veterinary Instruction . . . . . . . . .
Expenditure of the College . . . . . . . . . .
General Remarks . . . . . . . . . . .
Report on the Administration of the Glanders and Farcy Act in Calcutta and
Suburbs . . . . . . . . . . . . . .
Report of the Raymond Research Laboratory . . . . . . .
APPENDICES.
Table I . . . . . . . . . . . . . .
Table XIV-A . . . . . . . . . . . . . .
PAGES.
1
1—3
3
3
3-4
4-6
7
7
ANNUAL REPORT
OF THE
BENGAL VETERINARY COLLEGE
(Including the Reports of the Research Laboratory and the Glanders and
Farcy Department in Calcutta and neighbourhood)
FOR THE YEAR 1908-1909.
GENERAL ADMINISTRATION.
1. On the 1st April 1908, the mufassal portion of the Civil Veterinary
Department, Bengal, was handed over to Mr. D. Quinlan, as Superintendent,
while the Bengal Veterinary College, the Research Laboratory, and the area
under the Glanders and Farcy Act in Calcutta, Howrah, Alipore, and parts of
the 24-Parganas remained in my charge. I also exercised general control over
promotion and posting of Departmental Inspectors and Veterinary Assistants
of the Province.
VETERINARY INSTRUCTION.
2. The students.—The roll at the commencement of the session included
149 men (71 in Class "A", 48 in Class "B" and 30 in Class "C"). This
number was reduced at the close of the session to 132 (54 in Class "A", 48 in
Class "B" and 30 in Class ''C") comprising 102 Hindus, 23 Muhammadans,
four Budhists and three Native Christians. Of these, 53 came from Bengal, 14
from Bihar, two from Orissa, 53 from Eastern Bengal and Assam, two from
the United Provinces of Agra and Oudh, two from Madras, five from Burma and
1 from the Punjab.
Class ''A".—There were 71 admissions in Class A. Seventeen students
were subsequently removed or went away for various reasons, leaving 54.
Fifty-one were allowed to go up for the examination, three being stopped at the
Preliminary Test Examination.
Class "B".—Forty-eight
students were in this class throughout the year;
they all appeared before the Examiners.
Class "C''—This
class contained 30 students throughout the year, of
whom 29 appeared before the Board of Examiners, one being stopped at the
Preliminary Test Examination.
3. The Board of Examiners.—The Government of India have sanctioned a
permanent Board of Examiners for this college, composed of the Inspector-
General, Civil Veterinary Department as President and the Superintendents,
Government Cattle Farm, Hissar and Civil Veterinary Department, Eastern
Bengal and Assam, as members; they have fixed the dates of the examinations
and have allotted the subjects to each member of the Board. Owing to the
Inspector-General's inability, through illness, to attend, the Examining Board
was formed of the Superintendent, Government Cattle Farm, Hissar, as
President and the Superintendent, Civil Veterinary Department, Eastern
Bengal and Assam, and the second Superintendent, Civil Veterinary Depart-
ment, United Provinces of Agra and Oudh, as members. The examinations
began on the 1st March and lasted till the 9th.
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SECTION III.—REPORTS UPON PATHOLOGICAL AND OTHER SPECIMENS RECEIVED
FROM THE COLLEGE HOSPITAL.
During the year samples of blood, mucus, pus, parasites, tumours,
and other pathological specimens received from the college hospital were
examined for diagnosis.
SECTION IV.—POST-MORTEM EXAMINATIONS.
Post-mortem examinations were held in almost all the cases that died in
the General and Contagious Diseases Hospitals, and were demonstrated to the
senior students of the college.
Several interesting pathological and parasito-
logical specimens were collected.
SECTION V.—REPORTS UPON PATHIOLOGICAL AND OTHER SPECIMENS SENT BY
VETERINARY ASSISTANTS AND OTHER PERSONS IN MUFASSAL.
A considerable number of pathological specimens were received for
report from the Civil Veterinary Department and other sources.
SECTION VI.—STORAGE AND ISSUE OF BIOLOGICAL PRODUCTS.
We issued from the storage cellar the following sera and biological
products:—
Anti-Rinderpest Serum
„ Anthrax
Mallein
Tuberculin
...
...
...
...
Calcutta
Doses.
... 2,956
... 280
... 691
...
32
Muffassal
Doses.
21,851
...
90
...
SECTION VII.—RESEARCH AND OTHER SPECIAL WORK.
An outbreak of Lymphangitis amongst cattle has been of interest. This
disease is characterised by formation of primary and subsidiary abscesses,
implication of lymphatic glands, occurrence of lameness and pain in the legs
and feet. Its duration sometimes extends from four to six months accompanied
by loss of condition, ending sometimes in death. This disease differs very
little from one observed by Monsieur Wryburg in the Island of Sumatra and
described in the Bulletin de L' Institut Pasteur of 15th July 1907.
Examination of Bonemeal.—At the request of the Inspector, New Zealand
Department of Agriculture, several samples of bonemeal collected from the
Chingrighatta and Bengal Bone Mills were examined bacteriologically. The
results of the investigation indicated that those samples did not contain any
germs which are dangerous to higher mammalia.
Supply of material.—We have been able to continue our supplies of
fresh cultures of Bacillus Mallei and Anthrax to the Imperial Bacteriologist
for his work. Major W. D. Sutherland, M.D.,I.M.S., who has been deputed to
the Calcutta Medical College to study the methods of distinguishing different
kinds of mammalian blood, has been supplied with material whenever he requir-
ed it.
Post-graduate Courses.—Four officers of the Provincial Service underwent
a modified post-graduate course in Pathology and Bacteriology, three from
1st April to 30th June 1908 prior to promotion to the grade of Deputy Super-
intendents and one from 12th October 1908 to 12th February 1909.
Definite proposals and estimates for regular post-graduate courses of
study in this laboratory have been submitted to Government.
GENERAL REMARKS.
5. My thanks are due to Mr. S. N. Mitter, G.B.V.C., to Babu Sachindra
Kishore Das Roy, and to Babu Sasi Bhushan Mandal, the Laboratory
Artist, for their help during the year.