NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
GLOBIDIOSIS IN INDIAN CATTLE, WITH A DESCRIPTION OF NEW SPECIES.
179
had a patch of eczema about 4 in diameter on the right side of the hump, which
had become almost bald. This skin disease was reported as not yielding to any
treatment. Mange mites were not seen in skin scrapings, which were examined
under a microscope after treatment with 10 per cent. caustic potash.
Case No. 5.—A cow about 8 years old. This first came under the observation
of the Veterinary Assistant in charge, Farrukha-nagar Veterinary Hospital, District
Gurgaon, Punjab, when the animal apparently suffered from an attack of rinder-
pest and the Veterinary Assistant observed all the stages from fever to bloody
diarrhoea. The amount of blood in the faeces was so profuse as to make them al-
most red. The Veterinary Assistant suspected a complication of coccidiosis with
the rinderpest and procured some faeces, but having no other facilities in the dis-
pensary he put the faeces between two pieces of glass which he probably cut from
a glass pane. Although the material when it reached Lahore was nearly dry
yet a large number of spores were found in it on microscopical examination, and the
camera lucida drawings of the parasite are taken from this case. The cow in
question recovered from the illness within a fortnight, and at the end of this time
faecal smears were found negative for globidium spores.
NAKED EYE EXAMINATION.
The appearance of the affected mucous membrane varied according to the
number of cysts and period of infestation. In the majority of cases the lesions
consisted of whitish rounded or ovoid bodies, embedded in an area of opalescent
mucosa, slightly raised from its surroundings, the cysts being clearly visible. The
injected capillary blood vessels gave the pearly centre a rosy background. Pete-
chiae, ecchymoses and haemorrhages of varying dimensions were also not uncom-
mon and when a cyst was situated in these patches its appearance was rather obs-
cure. In a few places where the manifestations were discrete, the mucous mem-
brane appeared practically unchanged, except for a slightly raised eminence.
There were also seen erosions and thickenings and some fibrous connective tissue
in circumscribed small patches, which were probably the relics of old colonies.
The size of the cysts differed according to their age ; the smallest which were
round had a diameter of about 0.4 mm., whereas full grown ones, which were
oval or oblong, measured about 1 mm. in length and 0.8 mm. in breadth.
By exerting uniform pressure on all sides an individual cyst could be made to
stand out and then by tapping with a probe on the top the pressure of fluid could
be demonstrated. With a little more pressure a young cyst could be shelled out,
but in the case of a mature cyst, pressure causes it to burst, setting free an opaque
fluid containing the spores.
180 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, II
MICROSCOPICAL EXAMINATION.
For the purpose of diagnosis during life it appears necessary that several
examinations of the faeces should be made. As the spores and certain other stages
in the development of the parasite, which constitute the only diagnostic elements
present in the faeces, are not easily seen even microscopically in unstained prepara
tions, staining is necessary and the selection of the material is another important
point. In the case of apparently normal faeces scrapings from all sides of the drop
pings may be tried, but the blood stained part is probably the most suitable.
As the faecal specimens received were almost dried up, these were emulsified
in normal saline solution, from which were made fresh smears, which revealed the
spores. The latter were also found by Sheather's sugar floatation method for
worm ova.
With regard to the method of staining, Giemsa's stain was found to be the
most suitable. All other Romonowsky stains, Mann's double stain, Haematoxylin
and Eosin, or Carbol fuchsin and Methylene blue will also serve the purpose.
The contents of a colony were found to be composed of spherical protoplasmic
bodies varying in size. The smallest of these, which were rare, had only one nucleus,
whereas the larger ones, measuring from 60 to 90 μ in diameter, had as
many as 48 or more nuclei in a single optical plane. It appears that during the
development of the parasite, which presumably first remains intracellularly in the
gastro-intestinal epithelial cells, there is some reaction at the site of invasion,
and although the parasite may gain the upper hand, yet the tissues succeed in
forming a thin capsule around the colony. The protoplasm of the degenerated
cells gives place to an opaque watery fluid which surrounds the cysts and due to this
the colony becomes intercellular and is pushed more towards the sub-epithelial
connective tissue. As the maturation of single cysts proceeds, the thinning of
the envelop occurs and the internal pressure is raised. Rupture then occurs due
to pressure and the cysts are liberated in the lumen of the organ. The above
explanation appears to be necessary to account for the presence of numerous
spherical bodies in the faeces, in the absence of any extensive ulceration in the
mucosa of the gastro-intestinal canal. The haemorrhages probably occur in
the congested patches owing to irritation with rough food or to some bacterial
agency. As a complication of rinderpest no explanation for the aggressiveness of
the lesions is necessary, for this observation has already been made by various
authors in connection with the resuscitation of Coccidia in such cases.
In the case of the cyst itself the centrally situated nucleus, which is imme
diately surrounded by a hyaloplasm and then by a finely granular mass of pro
toplasm and finally by a wall, gradually grows bigger and undergoes division,
so that in a mature cyst there appear globules or corpuscles in the centre, around
which there remains a space which was previously occupied by the fluid and the
wall appears to have a double contour. Before rupture takes place, the nuclei,
Plate IX.
Ind. Jour. Vety. Sci. & Anim. Husb. Vol. V, Pt. 2.
Globidium fusiformis n. sp.
1. Individual spores.
2. A stage in the development of
the spores.
3. A lymphocyte.
4. Red blood corpuscles.
GLOBIDIOSIS IN INDIAN CATTLE, WITH A DESCRIPTION OF NEW SPECIES.
181
which are the precursors of the spores, become arranged around the periphery of
the cyst. The nuclei vary in size and around them there is a fluid which fills the
space and gives rise to a fluctuation in the contents of the cyst, when pressure is
exerted. Finally, there appear projections at the periphery of the spherical body
which radiate outwards from the centre. These represent the extremities of mar
ginally situated spores. In a freshly ruptured cyst as many as 20 or more spores
have been seen lying parallel to one another, in which position they remain for
some time due to attachment by the cytoplasmic debris between them.
The spores are elongated, spindle-shaped and slightly curved bodies, measur
ing about 13 μ in length and from 2 to 2.5 in width. In a stain smear of faeces,
when red and white blood corpuscles are also present, the microscopical picture
does not differ much from a blood smear from a case of trypanosomiasis.
These spores are tapering at both ends, one being bluntly rounded, and the
other is finely pointed, looking more or less like a tail. The nucleus is seen towards
the rounded end. It is vesicular in type and oval in shape measuring 3.25 × 2.25 μ.
In almost the centre of the spore there is a round chromatin mass constantly
occurring, which measures about 0.5 to 0.8 and takes an intense stain. The dis
tribution of the remainder of the chromatin in the spores is rather irregular and in
the form of a few granules occurring on both sides of the nucleus and varying
in number in individual spores. No granules showing amyloid reaction to iodine
were seen and no motility could be detected in fresh hanging preparations suspend
ed in dilute carbol fuchsin or normal saline solution. These preparations were
examined under a 1/6th objective and No. 4 occular and under dark ground
illumination. A cuticular covering is appreciable in some preparations stained
by Giemsa.
DISCUSSION.
Although the greater part of the life-history of the Globidium under report
is still unknown, it seems probable from a comparison with the description of the
spores and cysts of other forms, that it belongs to a species hitherto undescribed.
It is, therefore, proposed to refer to it as Globidium fusiformis, n. sp., although it
resembles in some respects G. faurei (G. gilruthi), a species common in the abo-
masum of the sheep in England, France and Australia. The chief points of differ
ence lie in the size, shape and form of the spores and in the behaviour of the spores
and cysts.
According to Triffitt (1925-27), who confirmed and elaborated Chatton's
description of G. gilruthi, the mature spores of G. gilruthi measure approximately
12 μ in length. One end remains bluntly rounded or cone-shaped, but the other
is prolonged to form a rostrum like process often being truncated at the tip. The
body is curved, flattened and frequently slightly twisted. In the case of G.
fusiformis the spores have not been seen with twisted and flattened bodies nor
182 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, II
is one end truncated. The average length is about 13 μ. These spores, as already
described, have elongated bodies and both ends tapering, the longer one of which
is so finely drawn out that the individual appears somewhat like a trypanosome.
The early schizont stages in this parasite are also intracellular, but before the
maturation of the spores the gravid epithelia probably dissolve allowing the cysts
to come in contact with one another while still encapsulated. When the capsule
ruptures the cysts are set free and those already mature burst and thus both
cysts and spores are found free in large numbers in the lumen of the alimentary
canal of the host.
As regards G. gilruthi, quoting Triffitt again, " The cysts occur only in the true
stomach or abomasum where they lie embedded in the mucosa ". She further
says, "On two occasions, in very heavy infections, cysts were found free in the
lumen of the stomach and the upper portion of the small intestine. It is possible,
however, that these had been discharged from the mucosa in the handling and
washing of the stomach, as, in subsequent examination of the surface of the mu
cosa and contents of the unwashed specimens no free cysts were found ".
G. gilruthi was first observed by Moussu and Marotel [ 1902 ], who mistook
it for a stage in the development of Eimeria faurei. Gilruth [ 1910 ] described
it as a common parasite of sheep in Australia, while Chatton [ 1910 ], who also
found it commonly in the slaughter houses of Paris, created a new genus, Gastro-
cystis, for it.
Further work was done on this group of parasites by Besnoit and Robin
[1912 ] and by Franco and Borges [ 1916 ], the latter of whom suggested that a'
new genus Besnoitia be created, but Nöller [ 1920 ], after collecting all the data
available regarding these parasites by different authors under various names,
placed them in one genus, Globidium, G. leucharti being taken as the type species.
Later, however, Henry and Masson [ 1932 ], who studied a similar protozoon
in the camel came to the conclusion that G. gilruthi must lapse as a synonym for
G. faurei, and that there are only four good species:—G. leuckarti, the type
species of the genus occurring in the horse, G. faurei in the sheep and goat, G.
besnoiti in the connective tissue of the ox and G. cameli in the camel. They define
the genus as large coccidia, the development of which comprises on the one hand
large oocysts with thick walls (formed by the fusion of large refractile corpuscles
or globules) and, on the other hand, endogenous reproductive forms which attain
a large size.
In conclusion, it may be observed that the following points are worthy of con-
sideration by Indian workers :—
1. That whatsoever may be the biological position of G. fusiformis, n. sp.,
it is obvious that it is liable to prove harmful, either directly or by exposing the
alimentary canal to other infections.
Ind. Jour. Vety. Sci. & Anim. Husb. Vol. V, Pt.
2.
Plate X.
Globidium fusiformis
n. sp.
1. A freshly ruptured globidium cyst.
2.
Particles of
fæces.
3. Red blood corpuscles.
4. Laked blood.
GLOBIDIOSIS IN INDIAN CATTLE, WITH A DESCRIPTION OF NEW SPECIES.
183
2. That although there is not as yet sufficient evidence to calculate the
economic loss, yet as one of the causes of debility in all cattle either bred for
milk or meat, the organism must be playing an important part.
3. That in this country where rinderpest and other weakening diseases are
so common, the resuscitation of this organism after an attack of one of the conta-
gious diseases should be kept in mind.
It seems possible that this organism may have a much wider distribution in
India than has yet appeared, and that its position may be the same as that pre-
viously held in this country by coccidiosis, regarding which the following remarks
were made in the Annual Report of the Imperial Institute of Veterinary Research,
Muktesar, for 1923-24 :—
" It is rather extra ordinary that the significance of this infection had not
been appreciated before in India, for examination at the Laboratory has shown
that nearly all cattle brought in from the vicinity are carriers of coccidia in
their alimentary tract".
Acknowledgments. I owe a debt of gratitude to F. Ware, Esquire, F.R.C.V.S.,
I.V.S., Director, Imperial Institute of Veterinary Research, Muktesar, for his kind
advice in arranging this paper, and I have also to express my indebtedness to
T. F. Quirke, Esquire, I.V.S., Director of Veterinary Services, Punjab, for his kindly
providing facilities for examination of the material through the District Veterinary
staff of the Punjab. Lastly, my thanks are due to H. Mohammad Hanif, Artist,
Lahore Veterinary College, for the camera lucida drawings.
REFERENCES.
Besnoit, C. H., and Robin, V. (1912). Sarcosporidiose cutanee chez une Bache,
Revue Veterinaire, 11, 649.
Chatton, E. (1910). Le Kyste de Gilruth dans le muqueuse stomacale des ovides.
Arch. Zool. Exper. et. Gen., 5 me. Ser. Vol. V, Notes et Revue, 114.
Flesch, M. (1883). Über ein Sporozoon beim Peerde. Zool. Anzeiger, 6, 396.
Franco, E. E., and Borges, I (1916). Sur la sarcosporidiose Bovine. Arquivos
Inst. Bact. Camara Pestana, 4, 269.
Gilruth, J. A. (1910). Notes on a Protozoon parasite found in the mucous mem-
brane of the abomasum of a sheep. Bull. Soc. Path. Exot., 3, 297.
Henry, A. and Masson, G. (1932). Considerations sur le genre Globidium cameli,
n. sp. parasite du Dromadaire. Annales de Parasitologie, 10, 385.
Nöller, W. (1920). Globidium (Gastrocystis Besnoiti) Handbuch der pathogenen
Protozoen, 2, 919.
Triffitt, M. J. (1915). Observations on Gastrocystis gilruthi, a parasite of sheep in
Britain. Protozoology, a Supplement to the J. Helminth, 1, 7.
—(1927).
Further observations on the Development of Globidium gilruthi-
Protozoology, a Supplement to the J. Helminth, 4, 83.
MINERALS IN PASTURE GRASSES IN INDIA
BY
A. VISWANATHA IYER, B.A.,
Assistant Physiological Chemist, Animal Nutrition Section, Bangalore.
(Received for publication on 13th March 1935.)
Largely through the stimulating lead given by Dr. Orr, Director of the
Rowett Institute, work on the mineral content of crops has been taken up and is
being prosecuted vigorously all over the world. In India practically every
agricultural institute is collecting some data on the subject.
The question was taken up by the Nutrition Section in a tentative form some
years ago. The aim was, and is, to grow and to determine the mineral content of
certain pure species of grass at a number of selected stations representing some
typical Indian soil and climate conditions.
The chief difficulties have been in connection with the establishment of
permanent cultures. In some places one species, in other places another species
is found to be difficult to grow. It has also occurred that after having been estab-
lished a plot may retrogress and eventually fail.
The scope of this work has been extended steadily so that there are now ten
Stations well distributed over the country at which plots have been laid down.
The present paper deals with the crops of two successive years from three
Stations only, namely, Pusa, Sabour and Bangalore.
The procedure has been to take three cuttings from each plot to represent
respectively, the young (just before flowering), the prime (full flower) and the
ripe (seed ripe and falling) stages of maturity. The sample including leaf, stalk
and heads is chaffed as a whole and milled.
The analytical data obtained from these samples will be considered from
various points of view.
I. INFLUENCE OF DEEP CULTIVATION ON THE MINERAL CONTENT OF GRASSES.
Table I shows the significant figures obtained with the Bangalore sample
It will be noticed that Pennisetum cenchroides and Cynodon Dactylon gave
roughly similar values for the two seasons. Therefore the climatic differences
that there may have been were not sufficient to produce marked mineral differ-
ences in these two species. In contrast to the results with Pennisetum cenchroides
and Cynodon Dactylon, Andropogon contortus gave entirely different results in the
2 seasons, the 1932 crop was rich in nitrogen and minerals—the 1933 crop was poor.
( 184 )
MINERALS IN PASTURE GRASSES IN INDIA.
185
It is a fact that the Andropogon contortus crop of 1932 was a new plantation,
the first year's growth after deep cultivation. The richness of this crop must,
therefore, be attributed to the effect of cultivation. Further tests are necessary
to corroborate and extend this important observation, which warns us that the
mineral content of a crop may be entirely misleading if the cultivation factor is
overlooked.
In the present enquiry the practice has generally been to establish a plot as
permanent grass and to sample this undisturbed permanent growth.
II. INFLUENCE OF SOIL AND OF GRASS SPECIES ON THE MINERAL CONTENT OF
GRASSES.
In Table II average values of the three cuts of each sample are given, the
discussion being confined to lime and phosphoric acid.
Certain characteristics of soils seem to be revealed by these figures. In every
case the Pusa crop contains more lime and less phosphoric acid than the average
value for the crop. Bangalore yields average phosphoric acid content and low
lime content. These results agree very closely with the known characteristics
and chemical constitutions of these two soils, the Pusa soil being a highly cal-
careous loam with low available phosphoric acid and the Bangalore soil a red
laterite deficient in lime and tending to be acidic.
With regard to phosphoric acid, it seems to be worthy of note that although
all the grasses grown at Pusa have a phosphoric acid content which is below
the ave age, yet the figures are not as low as we might have been led to expect
from the soil analysis data under the head available phosphoric acid.
Certain very interesting characteristics of grass species are also indicated by
the data of Table II.
Andropogon contortus appears to develop with relatively low amounts of
lime and phosphoric acid in its constitution. Cynodon Dactylon invariably
shows a considerable excess of lime over phosphoric acid, while Pennisetum cench-
roides is remarkable in containing an excess of phosphoric acid over lime, except
at Pusa, where, apparently, this tendency is masked by the highly calcareous
nature of the soil. Plants containing an excess of phosphorus over calcium are
apparently rare. The author has only noted two such cases up to date, namely,
Pennisetum cenchroides recorded here and Sorghum vulgare recently reported on
in another paper [Viswanatha Iyer, 1935]. These observations are considered
to be tentative only at present.
NITROGEN AND POTASH IN GRASSES.
The figures for nitrogen and potash in Table III show that there is a
parallelism in the variations of these two elements. Both are liable to fluctuate
considerably and both fluctuate in the same direction. For example, if, owing
to seasonal effect, the nitrogen content is above normal, the potash will also
be above normal. In contrast, the fluctuations of lime and magnesia are
186 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, II
relatively negligible. These elements remain almost constant while great
variations in the nitrogen and potash occur.
INFLUENCE OF MATURING ON MINERAL CONTENT.
Table IV gives the mineral composition of cuts made at different stages of
maturity.
Of the three grasses, Cynodon Dactylon gives the highest average nitrogen
content. It maintains its nitrogen most satisfactorily during maturing, Pennise-
tum cenchroides comes next in this respect while Andropogon contortus is definitely
inferior when judged by this important quality. The nitrogen content of this grass
falls to a very low level indeed in the course of maturing. It may be noted that
the phosphoric acid generally varies in the same way as the nitrogen and the
potash, but to a smaller extent.
Taking the figures as a whole it appears that maturing exerts a considerable
influence on per cent. of nitrogen and potash in the grass, a smaller influence
on the per cent. of phosphoric acid and only a minor effect on the per cent. of lime
and magnesia.
*
*
*
*
SUMMARY.
The mineral composition of three species of grass grown under different soil
conditions has been studied. The effect of the following factors was noted :—
(1) Influence of soil.—It was found that distinct differences in mineral
contents of the grasses could be attributed to the soil. (For example,
the plants grown at Pusa contained more lime and less phosphoric
acid than the average.)
(2) Influence of species.—It was found that each species of grass showed a
tendency towards a mineral make up peculiar to itself. (For
example, Cynodon Dactylon invariably showed a considerable excess
of lime over phosphoric acid, while on Pennisetum cenchroides the
tendency was for phosphoric acid to exceed the lime).
(3) Influence of maturing.—Maturing was found to exert a considerable
influence on the per cent. of nitrogen and potash in the grass, a
smaller influence on the per cent. of phosphoric acid and only a
minor effect on the per cent. of lime and magnesia.
(4) Influence of cultivation.—There is an indication that cultivation is likely
to effect the mineral content of herbage.
Acknowledgment. My thanks are specially due to Dr. Warth, the Physi-
ological Chemist to the Government of India, Bangalore, for suggesting this
work and for many helpful criticisms during the course of the work. My
thanks are also due to the Imperial Economic Botanist, Pusa, and the
Economic Botanist to the Government of Bihar and Orissa, Sabour, for laying
out the grass plots at the respective places, their maintenance, and the col-
lection of the samples at the proper time.