NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
222
Rhinosporidiosis in Equines
The following are the available details about the animal :—
District .
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Sub-division .
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Village .
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Breed and sex .
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Age
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Nature and size of growth . . .
Symptoms . . . . .
Date when first seen . . .
Date when growth removed . .
Subsequent history . . . .
Sambalpur.
Bargarh.
Bargarh.
Country-bred chestnut mare.
14 years.
Soft about 1" long and ½" high.
Slight blood-stained mucous discharge
from the affected nostril.
14th February, 1937.
15th February, 1937.
Not known, the animal being sold off.
The animal was brought to the hospital as it had audible breathing and
there was a slight bloodstained mucus discharge from one of the nostrils. The
Veterinary Assistant Surgeon while examining the affected nostril found a
small growth in the anterior part of the nasal cavity and he removed it.
The growth was about one inch long and half inch high and on naked eye
examination, it presented a cauli-flower-like appearance with a soft irregular
surface. When examined under a hand lens, it presented numerous pale
white specks embedded in soft tissue, which on histological examination proved
to be cysts or sporangia of Rhinosporidium.
Histologically the growth appeared to be identical in every respect with
the one described by Krishnamurti Ayyar. It was covered with stratified
squamous epithelium which presented an irregular outline and was in places
thrown into folds. In the superficial layers of the underlying highly vascular
sub-epithelial connective tissue were present large numbers of sporangia in
various stages of development. They were either oval or spheroidal in outline
and each was enclosed in a capsule which was thicker in the young than in the
older sporangia.
Inside each sporangium were present numerous spores in various stages of
development, the fully formed ones being usually in the centre and the small
ones at the periphery. In a few the fully-formed ones were situated at one
pole and the small ones at the other. The spores were enclosed in a thick
chitinous envelope with granular bodies inside them. Those bodies are consi-
dered by some to be refrangent spherules and by others to be sporules or
sporozoites.
As the sporangia grew older, they increased in size and ultimately pressed
upon the overlying epithelium which gradually diminished in thickness and
ultimately burst along with the capsules of the sporangia, thus enabling the
spores to get discharged on the surface of the nasal mucous membrane.
The case described in this paper is the second of its kind to be discovered
in a horse in India. Careful enquiries failed to reveal the existence of more
cases in the locality.
Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Pt. III]
PLATE XIV
A mature sporangium rupturing to discharge the spores with other
sporangia in various stages of development.
LAKSHMI
SAHAI
223
In view of the close relationship between Rhinosporidium equi and Rhinos-
poridium seeberi, the causal organism of Rhinosporidiosis in man, enquiries
were made to ascertain if human Rhinosporidiosis had been recorded in the
locality but the author learnt that the Medical Department was not aware of
the existence of the disease in Bihar. There is, however, a record of one case
of human Rhinosporidiosis in the Patna General Medical Hospital and two
in the Cuttack district of Orissa. In view of the fact that the disease has now
been shown to exist in this part of the country, besides Madras Presidency,
systematic search will probably reveal the existence of the disease in other
parts of India also.
REFERENCES
Ayyar, V. Krishnamurti (1927). Trans. of the Far
East. Ass. of Trop. Med.
Seventh Congress held in India, 3, 658-664.
––––––––––(1932).
I.J. Vet. Sci. & Anim. Husb. 2, 1, 49-52.
Cordero, E. H. and Vogeslang, E. C. (1928). Quoted by V. K. Ayyar (1932).
Quinlan, J. and de Koch, G. (1926). Quoted by V. K. Ayyar (1932).
Zschokke, E. (1913). Quoted by V. K. Ayyar (1932).
THE OCCURRENCE OF CORYNEBACTERIUM EQUI IN A
SHE-BUFFALO
BY
V. R. RAJAGOPALAN, G.M.V.C.
AND
V. R. GOPALAKRISHNAN, G.M.V.C.
Imperial Veterinary Research Institute, Mukteswar-Kumdun
(Received for publication on 25th March 1938)
(With Plate XV and one text-figure)
CORYNEBACTERIUM EQUI was first described by Magnusson [ 1923 ] as
a possible etiological agent in specific pneumonia in foals and he recorded
twelve cases of its incidence in a breeding establishment in Sweden. He
also conducted experiments which indicated that only equines and pigs were
susceptible to infection with the bacillus responsible for pneumonia in foals.
Subsequent workers have also reported the occurrence of the disease
in equines in other countries ; Meissner and Wetzel [ 1923 ], Lutje [ 1923 ],
and Lund [ 1924 ] in Germany ; Bull [ 1924 ] in Australia ; Dimock and
Edwards [ 1931 ] in America and Rajagopalan [ 1937 ] in India.
This organism has so far been recorded as occurring exclusively in the
equine species. The object of this paper is to record the occurrence of an
organism indistinguishable from Corynebacterium equi in its morphological,
cultural, biochemical and pathological characters in the genital tract of a
she-buffalo, the subject of abortion.
We are indebted to Mr. Tulsi Ram, Veterinary Assistant Surgeon on
special duty at Okara, for the material from the she-buffalo. The material
was received as swabs taken from the os uteri. The following history was
supplied with the material:
"Buffalo No. 92/4-3 in first calving—aborted about eight days ago—eight
months pregnancy—purchased from outside in pregnant state. Symptoms :
After-birth came out normally after abortion—discharge scanty, but purulent
and offensive—swab taken from vaginal wall near os uteri, as the animal was
badly straining and not properly secured."
At the same time and in the same place three other she-buffaloes also
aborted with the same clinical history.
As the case was suspected for bovine contagious abortion, the usual
cultural examination and small animal inoculation were done. On cultural,
biological and serological examinations, the material turned out to be negative
for evidence of infection with Br. abortus. But an Escherichia species and
a pigmented diphtheroid (C. equi) which is the subject of the present report,
were isolated.
( 225 )
226
Corynebacterium equi in a She-Buffalo
THE IDENTITY OF THE ORGANISM
Morphology.—Morphologically the organism grown on solid medium,
appears as short rods, mostly straight, occasionally slightly bent, with rounded
or clubbed ends. Most of them are small, almost ovoid, approaching the
spherical quite frequently. It is Gram-positive, non-motile, non-spore-forming
and non-capsulated. In broth-culture the bacillary form is more pronounced
than the coccoid.
Cultural characters.—Grown on plain agar plate, colonies are raised
and circular, with entire margin and moist and shining surface. They are
fawn-coloured and slimy in consistency. The colonies coalesce and tend
to run down and accumulate at the bottom of the slant. If stored at the
ordinary room temperature and exposed to light, after an incubation of
twenty-four hours at 37°C., the colour of the growth gradually deepens,
becoming salmon-pink.
In broth, growth is manifested by an uniform turbidity with a slight
granular deposit at the bottom which disintegrates completely on shaking.
The deposit develops a faint fawn colour after two or three days. Fre-
quently a ring of granular growth on the surface is also noticed.
It grows aerobically as well as under micro-aerophilic conditions, but
growth under anaerobic conditions, if any, was not noticeable.
The biochemical and other properties are tabulated below (Table I)
along with those of C. equi, for comparison :—
TABLE I
Tests
Adonite . . . .
Dextrose . . . .
Inulin . . . .
Maltose . . . .
Rhaffinose . . . .
Salicin . . . .
Xylose . . . .
Arabinose . . . .
Dulcite . . . .
C. equi.
—
—
—
—
—
—
—
—
—
Organism
isolated
—
—
—
—
—
—
—
—
—
Remarks
V. R. RAJAGOPALAN and V. R. GOPALAKRISHNAN 227
TABLE I—contd.
Tests
Glycerol . . . .
Lactose . . . .
Mannitol . . .
Rhamnose . . . .
Sorbitol . . . .
Galactose . . . .
Dextrin . . . .
Inositol . . . .
Laevulose . . . .
Mannose . . . .
Sucrose . . . .
Starch . . . .
Trehalose . . . .
Voges-Proskauer . . .
Methyl red . . . .
Indol production . . .
Nitrate reduction . . .
Hydrogen sulphide . .
Catalase . . . .
Litmus milk . . .
Gelatine . . . .
M.B. Reductase . . .
Potato . . . .
C. equi.
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
+
—
+
No change
No liquefaction
—
Moderate
dry
growth
red-
dish-yellow in
colour.
Organism
isolated
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
—
+
—
+
No change .
No liquefaction
—
Moderate
dry
growth
red-
dish-yellow in
colour.
Remarks
Readings taken
upto eleven
days.
15 days
228
Corynebacterium equi in a She-Buffalo
TABLE I— concld.
Tests
Growth on Sodium tellurite
medium.
Loeffler's medium . .
Blood agar .
.
.
.
C. equi.
Deep black .
Moderate growth
less abundant
than on agar.
Light pink.
No haemolysis .
Origanism
solated
Deep black.
Moderate growth
less abundant
than on agar.
Light pink.
No haemolysis.
Remarks
It is, therefore, evident that the organism isolated has characters identical
with those of C. equi.
SEROLOGICAL TESTS
(1) Agglutination.—When the material was first received, suspecting
the case might be one of abortion due to infection with Br. abortus—the
common causal organism of bovine abortion—a guinea-pig had been inoculated
intra-peritoneally with the material. This was bled twenty-seven days
later, and the separated serum was put to agglutination test against the or-
ganism under report as well as against Br. abortus. No agglutination took
place in any dilution in both instances.
Two rabbits were immunised, one with C. equi of foal pneumonia origin
and the other with the diphtheroid from the specimen. They were injected
intravenously with saline emulsions at four hundred millions per c.c. of the
respective organisms, three times at five days intervals. The dosage for the
successive injections was 0.25, 0.5 and 1 c.c. respectively. They were bled
on the eighth day after the last injection. The two sera were put to the
agglutination test against their homologous organisms. At the same time
they were put to cross-agglutination tests, the serum of the equine source
against the buffalo-strain of diphtheroid, and vice versa. No agglutination
was obtained in any of these tests. These tests were repeated in saline con-
centrations up to 1 . 5 per cent, still with negative results.
The fact, that the guinea-pig serum failed to reveal any titre for the
organism isolated and the sera of rabbits specially treated. for the purpose
also failed to give any agglutination, is not, however, very disappointing.
For, working with C. equi isolated from, cases of foal pneumonia, we have
never been able to demonstrate agglutinins in the naturally infected subjects,
and only occasionally in small animals intensively treated for the production
of agglutinating sera. This seems to have been the experience of most other
V. R. RAJAGOPALAN and V. R. GOPALAKRISHNAN 229
previous workers as well. Magnusson [ 1923 ] has reported his failure to
obtain agglutination of the bacillus with the blood of sick animals and
also from rabbits which had received repeated intravenous injections
of the culture. Bull [ 1924 ] also failed to demonstrate agglutinins in
a horse injected subcutaneously with an emulsion of the culture. Dimock
and Edwards [ 1931 ] however, claim to have produced agglutinating serum
(titre 100), by giving the horse four injections of culture with increasing
dosage at intervals of seven days.
(2) Complement fixation test.—As the agglutination tests were of no
avail in proving the identity of the two organisms, recourse was then had
to the complement fixation test, and cross complement fixation tests.
The sera from the two immunised rabbits, already mentioned, were used
as the source of amboceptors. The Kolmer's technique was followed in
these tests, using two full units of complement in one c.c. volume and 0.5 c.c.
quantities of each of the four other components.
The haemolytic serum was titrated using falling quantities of a 1 in 1,000
diluted serum, fixed quantities of complement, i.e., 0.3 c.c. and½ c.c. quantities
of a two per cent suspension of sheep red blood corpuscles. The mixture
was incubated for one hour at 37°C. and the titre at which haemolysis occurred
was read off. The stock haemolysin was then diluted for the test accordingly,
so that 0.5 c.c. contained two units of haemolysin.
Serum for complement was obtained from a healthy guinea-pig and was
diluted initially 1 in 30. Dilutions of complement from 0.1 to 0.5 c.c. with
a graded increase of 0.05 c.c. were made in normal saline solution to which
two units of haemolysin and two per cent suspension of sheep R. B. C, each
in half c.c. quantities were added and incubated for one hour at 37°C. The
smallest amount of complement giving complete haemolysis was noted and
the next higher, which is the full unit was taken as a guide to prepare a dilution
to contain two full units in one c.c. quantities.
Graded dilutions, in geometrical proportion, were made of the antigen,
for titration, from 2 to 32. Half c.c. quantities of a 1 in 10 dilution of a heated
healthy serum and one c.c. quantities of complement containing two full
units were added. After two hours' incubation at 37°C, haemolysin and
R. B. C. in required quantities were added followed by a further incubation
of one hour at 37°C. The smallest amount of antigen producing some in-
hibition of haemolysis was noted and the stock antigen diluted for the test
in such a way that one-third of the anti-complementary unit was contained
in ½ c.c. volume.
Having titrated the components the complement fixation test was con-
ducted using the sera of the two rabbits which had been immunised against
C. equi and the buffalo diphtheroid respectively. The sera were heated
overnight at 55°C. Dilutions of the serum from 1 in 10 to 1 in 800 were made,
and the titrated quantities of antigen and complement were subsequently
230
Corynebacterium equi in a She-Buffalo
added and incubated for two hours at 37°C. Then the requisite quantities
of haemolysin and sheep R. B. C. were added. The contents were mixed
well and incubated for one hour at 37°C.
A healthy rabbit serum was also tested against both the antigens for
control.
In addition, a complete set of the usual controls for the reliability of the
components were kept as follows:—Serum control with one-tenth dilution
of serum, complement, haemolysin and R. B. C, but with no antigen ; antigen
control with ½ c.c. of antigen, complement, haemolysin and R. B. C, but
with no serum; haemolytic system control with two units of haemolysin
complement and R. B. C, but with neither serum nor antigen; R. B. C.
control with two per cent sheep R. B. C. in normal saline only.
All controls were satisfactory.
The result is tabulated below (Table II).
TABLE II
Serum tested (after
heating at 55° C.
overnight
C. equi of foal
pneumonia origin
Do. .
Diphtheroid of buff-
cow origin.
Do. .
Healthy rabbit se-
rum
Do. .
Serum dilutions
1/10
+ + + +
+ + + +
+ + +
+ + +
+
+
1/50
+ + +
+ + +
+ +
+ +
—
—
1/100
++
+
+
+
—
—
1/200
—
—
—
—
—
—
1/400
—
—
—
—
—
—
1/800
—
—
—
—
—
—
Antigen
C. 13
C. 200
C. 13
C. 200
C. 13
C.200
Controls
Se r
um
—
—
—
—
—
—
Antigen
—
—
—
—
—
—
Haemolytic
system
—
—
—
—
—
—
R. B. C.
+
+
+
+
+
+
NOTE :—C. 13 = C. equi of foal pneumonia origin.
C. 200
= Diphtheroid of she-buffalo origin.
(—)
= Complete haemolysis.
.
.
.
No fixation of complement.
( + )
=25 per cent inhibition of haemolysis .
( + +)
= 50 per cent inhibition of haemolysis .
( + + +) =75 per cent inhibition of haemolysis .
( + + + +)= 100 per cent inhibition of
haemo¬
lysis
Degrees of fixation of comple-
ment.
PLATE XV
[ Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Pt. III.
The lung of Foal No. 417 that was infected with the buffalo strain of diphtheroid, showing
abscesses in the anterior portion of both the main lobes
V. R. RAJAGOPALAN and V. R. GOPALAKRISHNAN 231
It will be seen that the foal pneumonia organism fixes complement in
the presence of the buffalo-diphtheroid-serum at about the same titre as in
the presence of its homologous serum and vice versa.
PATHOGENICITY
The organism was found to be non-pathogenic to rabbits and guinea-pigs.
With a view to ascertain the pathogenicity of the organism for young equines,
Foal No. 417, aged fourteen days was swabbed inside both nostrils with an
emulsion of the buffalo strain of diphtheroid. The animal behaved exactly
like a foal infected with C. equi (Text-fig. 1). It showed a distinct rise in
temperature on the evening of the sixth day. The animal was dull and
lacking in appetite. It had an occasional cough and became progressively
weak. The respiration was distressed and gurgling noises could be heard on
auscultation of the lungs. The temperature continued to be high until just
before death when there was a slight fall. The foal died on the sixteenth
day after infection. A post mortem examination, the anterior portion of
both the main lobes of the lungs were found studded with characteristic
abscesses (Plate XV). An organism indistinguishable from C. equi was
re-isolated from the lung abscesses in pure culture.
Whether the organism is capable of causing abortion or purulent metritis
in she-buffaloes under experimental conditions could not be determined owing
to lack of pregnant she-buffaloes. In the hope that a pregnant cow might
react in a similar way, Cow No. 5, nine months pregnant, was instilled intra-
vaginally with five c.c. of a saline emulsion of the organism made from four
48-hrs. old agar slants, combined with an oral administration of an equal
quantity of a similar emulsion. Twelve days later the cow gave birth to
a calf which died soon after birth. The organism could not be recovered
from the uterine discharges of the cow, nor from, any of the internal organs
of the dead calf.
Another experiment was conducted to find whether this organism could
cause pneumonia or joint-ill in bull-calves, in the same way as C. equi does in
young equines. Two calves, aged two to three months, were swabbed in both
nostrils with an emulsion of the organism. No lesions attributable to in-
fection with C. equi developed in either of them.
232
Corynebacterium equi in a She-Buffalo
Fig. 1.
Clinical chart of Foal No. 417 (born on 26th June 1937) infected with the buffalo
strain of C. equi.
V. R. RAJAGOPALAN and V. R. GOPALAKRISHNAN 233
DISCUSSION
From the cultural and biochemical characters of the organism coupled
with its serological affinity to C. equi and pathogenicity to young foals, there
remains no doubt the organism under consideration is a strain of C. equi.
This is the first time that the organism has been recorded in a species
other than equine. It was isolated from the uterus of a she-buffalo that
had aborted. Following abortion there was a foetid purulent slimy discharge
from the uterus and the organism was obtained from this discharge. The
only other organism that was grown besides C. equi was a coliform organism
and the question naturally arises why the latter should not be held as the
cause of the condition. It is true that certain strains of coliform organisms
are capable of causing inflammatory conditions of the urinogenital tract
of man and animals. Such strains are highly virulent for the guinea-pig
and mice, causing, on fresh isolation, by intraperitoneal inoculation, a suppura-
tive or purulent peritonitis or a fatal septicaemia. But in this case, guinea-
pigs and mice that had received intraperitoneal injections of the material
developed no illness, and on destruction revealed no pyogenic lesions. It
would appear, therefore, that the coliform organism was a banal organism
devoid of pathogenic properties. There is, therefore, the probability that
the pyogenic lesions met with in the buffalo were due to the other organism
isolated namely C. equi. This organism is non-pathogenic to laboratory
animals, which fact should explain why the guinea-pigs and mice inoculated
with the purulent material from the buffalo containing the diphtheroid deve-
loped no lesions.
But the mere isolation of an organism from lesions does not definitely
establish it as the pathogen. Lesions similar to those in the buffalo could
not be induced in a pregnant cow by the intravaginal instillation of the isolated
organism. Owing to lack of pregnant buffaloes, the pathogenicity of the
organism to the species from which it was isolated could not be tried under
experimental conditions. It has not been possible, therefore, to advance
experimental evidence definitely incriminating this organism as the cause
of either post-abortion purulent metritis or of abortion although the cir-
cumstances of the case under report are strongly suggestive of its being the
cause of one or both. However, irrespective of its pathogenic significance,
the mere occurrence, in the buffalo, of Corynebacterium equi, an organism
which has so far been naturally met with exclusively in the equine species,
is of sufficient interest and is deserving of report.
SUMMARY
Corynebacterium equi has been recognised by several workers as a cause
of pneumonia in young equines. It has not so far been recognised as occurring
in any other species of animals. Its occurrence in pyometra following abortion
in a she-buffalo is, therefore, recorded in this article. Its identity with Cory-
nebacterium equi has been proved by morphological, cultural, biochemical,
234
Corynebacterium equi in a She-Buffalo
serological and biological tests. The probability of its being an etiological
factor of either abortion or of post-abortion pyometra is discussed.
REFERENCES
Bull, L. B. (1924). J. Comp. Path. Ther. 37, 294.
Dimock, W. W. and Edwards, P. R. (1931). J. Amer. Vet. Med. Ass. 79, 809.
Kolmer, J. A. and Boerner, F. (1931). Approved Laboratory Technique.
Lund, L. (1924). D. T. W. 32, 4.
Lutje, F. (1923). ibid, 31, 559.
Magnusson, H. (1923). Archiv.f. Wiss. w. prakt. Tierheilk, 50, 22.
Meissner, H. and Wetzel R. (1923). D. T. W. 31, 449.
Rajagopalan, V. R. (1937). Ind. J. Vet. Sc. & Anim. Husb. 7, 38.
PSEUDANOPLOCEPHALA CRAWFORDI BAYLIS, 1927.
BY
S. VAITHIANATHA MUDALIAR, G. M. V. C.
Assistant Lecturer, Madras Veterinary College
AND
K. S. GOPALAKRISHNA IYER, G. M. V. C.
Veterinary Assistant Surgeon, Madras Veterinary College.
(Received for publication on 30th March 1938.)
[With Plate XVI and two text-figures.]
INCIDENCE of adult cestodes in the pig appears to be very limited. The
cases on record so far are by Maplestone and Southwell [1923], Baylis [1927]
and Hall [1924]. Maplestone and Southwell [1923] described Paramoniezia
suis from an Australian wild pig. Baylis [1927] described Paramoniezia pha-
cochaeri from an African wart hog and Pseudanoplocephala crawfordi obtained
from a wild pig in Ceylon. Hall [1924] appears to have recorded an undeter-
mined Anoplocephalid tapeworm in a pig in America. It is thus seen that no
adult tapeworms have so far been reported from India and hence this record of
a tapeworm from this part of India. In the last week of December 1937, a col-
lection of helminths of the pig was made by the writers and the worm to be
described belongs to that collection. In the museum of parasitological collec-
tions of this College, about half a dozen specimens of tapeworms from the
pig collected by M. Anant Narayan Rao and L. S. Parameshwara Iyer during
1932 and awaiting identification were also examined. All the specimens
resemble, in almost all details, the parasite P. crawfordi Baylis, 1927. A
description of the parasites is given below :
The worms measure from 21 to 31 cm. in length and 3 to 4 mm. in
width. The segments are wider than long in the anterior portion while there
is a tendency to an increase in the length at the posterior end of the
strobila. The gravid segments are very much contracted.
( 235 )