NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
178
Vitamin-A Deficiency in the Diet of Farm Animals
an exposure for three days before cocking gave a hay with only 116±9 units
per grm. It was also found that cock cured and unstacked hay kept for
two and a half months contained 233±20 units per grm. but hay stacked for
forty-nine days had only 144±10 units [Woods et al, 1936].
Briefly stated, the provision of an adequate supply of vitamin-A in the
dietary of animals, particularly cattle can be accomplished only by making
their natural feed richer in carotene content and not by feeding vitamin-A
concentrates. In India their main feed consists generally of a roughage and
a concentrate ration, the feeding of carrots and other roots not being common.
The concentrate ration is generally composed of grains and oil-cakes which
cannot be made to yield appreciable amounts of vitamin-A. Hence animals
have to be provided with fresh green feeds, which under ordinary farm condi-
tions provide an adequate amount of carotene. As it may not be possible
to provide such a feed throughout the year on any large scale, the methods
of preservation of fodder plants without destruction of the vitamin-A potency
have to be investigated. Ensilage and hay-making are the two methods open
and considerable work has to be done with regard to these aspects of the
question. It should also be stated that by proper plant breeding, fodder
crops with improved carotene contents could be developed.
CONCLUSION
In the foregoing pages, a general review has been given of the carotene
contents of various stock feeds and the requirements of vitamin-A for the
proper maintenance of the health of live-stock. It would be appropriate
here to consider the question of avitaminosis-A under Indian conditions.
It has been shown elsewhere [Guilbert et al, 1934] that vitamin-A deficiency
in the diet of farm stock may occur both under stall feeding conditions as
well as when grazing and the common symptoms are—
(a) expulsion of the foetus prematurely or dead at term or birth of
weak calves with or without eye lesions and associated with
retention of the placenta,
(b) severe diarrhoea in weak new born calves, and
(c) ophthalmia in growing animals.
The existence of these troubles among the cattle of this country is well
known and it is probable that malnutrition due to avitaminosis-A is widely
prevalent in India. For instance, the incidence of abortion in various dairy
herds such as in Quetta, Wellington, etc., and the occurrence of the widespread
trouble of " blindness in calves " in Sind and Baluchistan have been shown
to be due to vitamin-A deficiency in the ration and these troubles have been
reproduced under controlled conditions. A supplement of green feed in the
ration prevented the occurrence of both the above troubles in Quetta.
K. C. SEN AND P. A.
SESHAN
179
It has been shown previously that the maintenance requirement of caro-
tene for average cattle may be considered to be about 14 mg. per day. A
recent analysis has shown that rice straw from three different parts of India
did not contain any carotene and the same was also true of various concen-
trate rations used in our dairy farms. Consequently with a diet based on
these concentrates and rice straw as the sole roughage, there is a gross defi-
ciency of vitamin-A in the diet. Various samples of hay have also shown negli-
gible amounts of carotene contents. The importance of choosing proper
diets including a good variety of green fodder is, therefore, quite evident
and needs no further comment. It is to be remembered here that every
green fodder is not an equally rich source of carotene, jowar, maize and some
varieties of pasture grass being much poorer than clover, lucerne or berseem.
In conclusion, it may be emphasised that the problem of avitaminosis-A
in cattle and especially in milch cows has far greater significance than what
appears on the surface. It has been pointed out already that the vitamin-A
content of milk is dependent on the quality of the feed given to the cow and
as milk is probably the most important source of vitamin-A for growing
children, avitaminosis-A in cattle is intimately connected with the Public
Health problem in India. This aspect of the question was dealt with in detail
by Colonel Sir Arthur Olver in his Presidential address to the Medical and
Veterinary Research Section of the Indian Science Congress in 1937 under
the title " The relation of animal nutrition to Public Health in India " and
no further elaboration is, therefore, required at present.
SUMMARY
This paper attempts to summarise the present position of avitaminosis-A
so far as it relates to the practical feeding of farm animals.
Cereals, grain feeds, vegetable oils and oil-cakes are found to be poor
sources of carotene (the precursor of vitamin-A) which is generally associated
with the pigments of all plant material.
Green young forage plants are, however, found to be satisfactory sources
of carotene for animals.
The vitamin-A requirements of rats, dogs, poultry, swine and cattle
are discussed. The excretion, storage and depletion of the vitamin-A reserve
of animals through reproduction and milk are described.
The point is stressed that the provision of suitable green feeds and pasture
lands is the only practical method of supplying the carotene requirement of
farm animals. In this connection, the importance of preserving the vita-
min-A potency of green plants when converted into hay or silage has also
been emphasised.
The existence of avitaminosis-A in a mild form under the ordinary condi-
tions of stock feeding in India and the need of rectifying this condition are
also considered.
180
Vitamin-A Deficiency in the Diet of Farm Animals
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—and
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Guilbert, H. R. (1934). Ind. Eng. Chem. Anal Edn. 6, 452-454.
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—and
Hart, G. H. (1934). J. Nutri. 8, 25-44.
—
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Hale, F. (1933). J. Heredity 24, 105-106.
—
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Hart, G. H. and Guilbert, H. R. (1933). Calif. Agri. Expt. Sta. Bull. No. 560.
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Hetler, R. A. (1934). J. Nutri. 8, 75-103.
Hilton, J. H., Hauge, S. M. and Wilbur, J. W. (1935). J. Dairy Sci. 18, 795-800.
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Iwanova, N. (1935). Problems of Nutrition, Moscow 4, No. 4, 134-141.
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Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Part III ]
PLATE IV
Babesia bigemina (Smith and Kilbourne).
All figures are drawn with the aid of Camera lucida and the magnification is Ca.
× 2,000.
FIGS. 1-3.—Represent variety of pictures which the red or pink staining material
presents when stained with Giemsa (one of the Romanowsky stains).
FIG. 4.—Shows the effect of Fuelgen's reaction on the nucleus counterstained
with light green. Note the structure of the nucleus at the apex and the
hydrolised vacuolated area at the broad pole of the parasite.
FIG. 5.—Shows a Fuelgen preparation counterstained with Heidenhain's iron¬
alum-haematoxylin. The hydrolised area is shown to have taken up the
haematoxylin stain.
ON THE NUCLEAR STRUCTURE OF BABESIA BIGEMINA
(SMITH AND KILBOURNE)
BY
H. N. RAY, M.Sc., PH.D. (LOND.)
Imperial Veterinary Research Institute, Mukteswar
(Received for publication on the 10th June 1938)
(With Plate IV)
THE family Piroplasmidae to which belongs the genus Babesia is placed
amongst sporozoa under the order Haemosporidia. But since the publica-
tion of Breinl and Hindle's [1908] work on Babesia canis, the consideration of
a probable relationship of the genus Babesia with the flagellate, has become
a matter of considerable importance from the systematic point of view. Find-
ing of flagellate forms at some stage of its development in the body of the
vertebrate host has, in a way, paved the way for putting forth the arguments
in favour of flagellate origin of this genus. Thomson and Fantham [1913]
who have successfully cultivated Babesia canis in vitro did not, however, come
across any flagellate stage as shown by Breinl and Hindle.
Dennis [1930] while working on the morphology of Babesia bigemina, the
causative organism of Texas fever, came to a somewhat similar inference
although actual flagellate forms were not encountered by him. An extra-
nuclear granule, or blepharoplast and its connection with the nucleus similar
to that described for Babesia canis has also been shown to be present in Babesia
bigemina except that in the former a flagellum was found to originate from the
granule while in the latter this organelle was absent. Presence of this extra-
nuclear granule or ' blepharoplast' was considered by Dennis a feature of
sufficient importance to suggest its flagellate affinity.
Smith and Kilbourne [1893] who first described the organism of Texas
fever did not give any account of the nuclear structure. Laveran and Nicolle
[1899] pointed out that the nucleus was situated at the broad pole of the para-
site and that it consisted of a ' karyosome ' with a clear area around it. Fan-
tham [1907] gives a detailed description of the distribution of the chromatin*
in this organism. His observations, however, were based on dried smears
stained with one of the Romanowsky stains, a method which cannot be
relied upon for studying the cytological details. Nuttall and Graham-Smith
[1908] describe the nucleus as a small granule with a row of granules extending
* I put this word in italics because any structure that stained pink or red with
Romanowsky's stain is usually described as chromatin without paying much attention
to its true chemical composition which according to Feulgen (1927) is mainly built upon
the thymonucleic acids as its base.
( 183 )
184
Nuclear Structure of Babesia bigemina
from it; these authors have also shown that the nucleus is situated at the
pointed end of the pyriform body. This is of course quite opposed to what
has been described by Laveran and Nicolle.
In order to determine what actually is the position of the Indian strain
of Babesia bigemina with regard to its nuclear structure, detailed observations
were made on wet fixed smears. The results of studies incorporated in this
article will at once reveal that the structure of the nucleus is quite different
from what has hitherto been described. The nucleus is confined to the point-
ed end of the parasite and consists of ' karyosome ', a nuclear membrane, and
finely granular chromatin. No indication of any extra nuclear granule or
blepharoplast was present in the preparations used for this study.
MATERIAL AND METHODS
Material for study was obtained from hill-bulls which showed heavy in-
fection with B. bigemina as a result of resuscitation due either to experimental
theileriasis or rinderpest. Wet blood smears were fixed in Bouin-Duboscq¬
Brasil's fluid, alcoholic Bouin's fluid, Schaudinn's fluid and Carnoy's fluid.
Smears were fixed for fifteen to twenty minutes except in Carnoy's in which
they were kept for five minutes. Stains used were Heidenhains iron-alum
haematoxylin, Giemsa and Mallory's triple stain. In order to determine the
distribution of chromatin in the nucleus, the method of Feulgen's nucleal reac-
tion was employed. It may be mentioned here that this reaction is based on
the hydrolysis of other constituents such as guanine and adenine combined with
thymonucleic acid by means of dilute hydrochloric acid at 60°C. and later on
obtaining a permanent colour reaction by treating it with basic fuchsine. By
this method, so far as is known at present, the proper seat of chromatin in a
nucleus can be ascertained with accuracy. Smears treated in this way were
counterstained with light green in ninety per cent alcohol. Some of the smears
instead of being counterstained with light green were stained in iron-alum¬
haematoxylin. This method of staining revealed a granule which was invisible
in Feulgen preparations. Various pictures of the parasites thus obtained were
compared and careful Camera lucida drawings were made in each case.
OBSERVATIONS
The routine method of staining with Giemsa or Lieshman stain showed
so-called chromatin at various places of the pear-shaped organisms and needs
little mention from the point of view that dry smears are useless for studying
the cytological details. In such preparations a deeply stained pink or red
granule appears at the broad pole of the parasite which was interpreted as
being a nucleus by Laveran and Nicolle [1908]. Besides this granule, however,
a deeply stained area is also seen at the pointed end of the parasite. This was
studied in great detail by Nuttall and Graham-Smith [1908] and it was pointed
out by them that the nucleus was, truly speaking, situated at the apical end of
the organism and consisted of a granule from which extend a number of small
granules after the manner of comet (Plate IV, fig. 1). As a matter of fact
H. N.
RAY
185
this is exactly the picture of the nucleus which one gets so often in dry pre-
parations stained with one of the Romanowsky stains. But it must be said
that this holds good only in cases where we are dealing with the typical pear-
shaped organism. In rounded or amoeboid forms a single deeply stained
granule is usually more prominent than others (Plate IV, fig. 2). In
preparations fixed in any of the fixatives mentioned above and stained in
iron-alum-haematoxylin shows only a dark dot at the apex of the parasite and
sometimes a dark thread-like structure can be seen extending from it. The
granule at the broad end which is two to four times as big as one at the apex,
also takes up a deep black stain, but in no case was this structure seen to be
connected with that thread. Dennis [1930] has also called this bigger granule
the nucleus and has shown that it is connected with the smaller one which
he refers to as the blepharoplast by means of a rhizoplast. Round this bigger
granule he has demonstrated the presence of a nuclear membrane—a structure
which unfortunately could neither be seen by Rees [1934] or by the author.
It will, however, be seen later on that the bigger granule is devoid of true chro-
matin and has, therefore, no connection with the nuclear apparatus of the
parasite.
In Feulgen preparations the substance contained in the bigger granule
at the broad pole is completely hydrolysed and appear as a vacuole when
counterstained with light green, while that at the apex gives a definite chro-
matin reaction (Plate IV, fig. 4). The nucleus in such preparations
appears to consist of a deeply stained karyosome usually situated at the ex-
treme narrow end and surrounded by a circle of fine granules. These fine
granules may sometimes take up a linear position keeping the karyosome
either at its apical, central or lower end. The number of these fine granules
varies and are arranged at the periphery of an achromatinic nuclear mem-
brane.
After Feulgen's reaction some smears were stained with iron-alum¬
haematoxylin. In these preparations the nucleus presented the same picture
as mentioned above except that the nuclear membrane was more deeply
stained. A very striking feature which was noticed in this preparation was
that the area completely hydrolysed by Feulgen reaction and appeared as a
vacuole, had taken up a deep black stain (Plate IV, fig. 5). Smears fixed
in Carnoy's fluid and stained either with iodine or Best's carmine, however,
revealed the true nature of this granule. With iodine it gave a pale brown
reaction, while with Best's carmine it appeared as a faintly pink area;
suggesting thereby that this area contained certain substance which is allied
to glycogen.
It is now clear, therefore, that the deeply staining granule at the broader
pole of the pyriform body has no relation with its nuclear apparatus, but
instead, is a product of metabolic activity of the parasite. This finding also
explains why this granule does not constantly occur in every individual para-
site at every stage.
186
Nuclear Structure of Babesia bigemina
The nucleus in Babesia bigemina as described above, is situated at the
narrow apical end of the organism and consists of a karyosome surrounded by
a circlet of fine granules, arranged at the periphery of the nuclear membrane.
With this data at our disposal we can no longer entertain the theory of flagel-
late origin of this sporozoan as propounded by Dennis [1930].
SUMMARY
The nuclear structure of the Indian strain of Babesia bigemina has been
studied in detail. The nucleus is found to consist of a karyosome surrounded
by a circlet of fine granules which give a positive Feulgen reaction. These
fine granules are arranged at the periphery of an achromatinic nuclear mem-
brane. The nucleus is situated at the narrow apical end of the pyriform
body. A granule which is sometimes seen at the broad pole of the parasite,
is two to four times the size of the karyosome and gives a negative Feulgen
reaction. With iodine and Best's carmine this granule gives a faintly posi-
tive reaction for glycogen.
REFERENCES
Breinl, A. and Hindle, E. (1908). Ann. Trop. Med. & Parasit. 2, 233-248.
Dennis, E. W. (1930). Univ. Calif. Publ. Zool. 33, 179-188.
Fantham, H. B. (1907). Quart. J. Micro. Sci. 51, 295-323.
Laveran, A. and Nicolle, M. (1899). C. R. Soc. Biol. 6, 748-751.
Nuttall, G. H. F. and Graham-Smith, G. S. (1908). Parasitol. 1, 134-142.
Rees, C. W. (1934). J. Agric. Res. 48, 427-438.
Smith, T. and Kilbourne, F. (1893). Bull. Bur. Anim. Indus., U. S. Deptt. Agri.
1, 177-304.
Thomson, J. G. and Fantham, H. B. (1913). Ann. Trop. Med. & Parasit. 7,
621-630.
PLATE V
[ Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Pt. III
FIG. 1.
Section of growth from nose of bullock showing
asci of rhinosporidium. The asci are in close
proximity of epithelium. × 60
FIG. 2.
Section of rhinosporidial growth from the nose
of a pony. × 60
FIG. 3.
Section of rhinosporidial growth from nose
of a man. × 60
FIG. 4.
Section of rhinosporidial growth, Bovine.
Note the ascus bursting through the
epithelium. × 60
RHINOSPORIDIOSIS IN BOVINES IN THE MADRAS
PRESIDENCY, WITH A DISCUSSION ON THE
PROBABLE MODES OF INFECTION*.
BY
M. ANANT NARAYAN RAO, G.M.V.C.
Lecturer in Parasitology, Madras Veterinary College, Madras.
(Received for publication on 22nd February, 1938)
(With Plates V-VIII)
CONTENTS
1. INTRODUCTION
2. SITE OF INFECTION
. .
. .
3. SEX AND AGE INCIDENCE
. .
. .
. .
. .
. .
. .
. .
. .
. .
4. DURATION OF THE DISEASE AND RECURRENCE AFTER
EXCISION
5. CONCURRENT INFECTION
6. CLINICAL FEATURES
. .
. .
. .
. .
. .
. .
. .
. .
. .
. .
. .
. .
7. HISTOPATHOLOGY AND LIFE-HISTORY OF THE ORGANISM
IN TISSUE
. .
8. IDENTITY OF THE PARASITE
. .
. .
. .
. .
9. DISSEMINATION OF SPORES FROM THE LESIONS
10. ATTEMPTS TO CULTURE RHINOSPORIDIUM
11. PROBABLE MODES OF INFECTION
12. SUMMARY . .
. .
. .
. .
. .
. .
. .
. .
. .
. .
. .
. .
. .
Page
187
189
190
190
190
190
191
192
192
193
194
196
1. INTRODUCTION
THERE is a remarkable paucity of record of the occurrence of rhinospori¬
diosis in animals. Only four reports are available from different parts of the
world, recording the occurrence of the disease in eight animals. Zschokke
[1913] found Rhinosporidium in the nasal growth from a horse in South
Africa sent to him by Theiler. Fourteen years later Quinlan and de Kock
[1926] found the parasite in the nasal growths of two mules in South Africa.
Cordero and Vogeslang [1928] reported their finding of the fungus in the nasal
*Paper read at the Jubilee Session of the Indian Science Congress, held in Calcutta,
January, 1938.
( 187 )