NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
ACTINOMYCOTIC DERMATITIS IN CROSS MERINO LAMBS
By H. K. LALL, B.Sc., M.R.C.V.S., Assistant Animal Husbandry Commissioner
with the Government of India and V. R. RAJAGOPALAN, Dip. Bact. (Vict.)
Indian Veterinary Research Institute, Mukteswar-Kumaon
(Received for publication on 4 November 1948)
(With plates I to III)
A PECULIAR form of dermatitis was noticed among the lambs at the Govern-
ment Livestock Farm, Hissar, and, at one time, it caused some concern as
mortality among the affected ones was rather high. The disease was observed in
1942 among a few lambs but it attracted particular attention when it assumed
serious proportion in 1944. The disease at first, occurred in one of the blocks of
the farm and spread to another one, about four miles away, in about two weeks.
Clinical features
The outbreaks occurred in the rainy months of July, August and September
and higher incidence was recorded, in pens which were damp, being in low lying
areas. The Hissardale—a breed evolved as the result of crossing Bikaneri with
Merino, possessing finer wool suffered more than the Lohis or Bikaneris.
Out of 268 Hissardale lambs 91 (34 per cent) got the disease and 26 (28.6 per-
cent) of the affected animals died. The disease among lambs of the Lohi breed
was relatively mild; 21 out of 170 lambs (12.36 per cent) were affected but only
one (4.8 per cent) died. No cases were recorded among the Bikaneri lambs. The
disease was confined to very young suckling lambs not above a fortnight in age
No adult sheep contracted the disease.
SYMPTOMS AND COURSE
The very early cases usually passed unnoticed. With the progress of the
disease, symptoms were more pronounced and the affected lambs showed dullness
and depression. The affected portion of the skin thickened and was hard to feel.
The parts usually involved were the skin of the back and the neck although some
times, scale-like lesions also developed around the eyes, nose, lips and ears. The
skin of the abdomen, inside of the legs and below the fetlock, was never affected.
The skin of the back was often extensively involved and moulded itself into a sort
of hard, rigid, inflexible, inelastic plate, the patient then becoming imprisoned in
a kind of plaster of Paris jacket. Because of its peculiar disposition, the skin of
the neck was wrinkled (Plate I, fig. 2) and the rigidity of the neck interfered with
suckling. The wrinkled skin sometime cracked and wounds developed which
invariably got infested with maggots, if left untreated (Plate I, fig. 1). The skin
on the back lost all wool and presented a raw appearance. In a number of cases,
nodules exuding yellowish pus could be seen (Plate II, fig. 1). In recovered cases,
2
ACTINOMYCOTIC DERAMATITIS IN CROSS MERINO LAMBS [Vol. XIX, Part I,
it took a long time, for the skin to assume normality and the wool remained dis-
coloured for quite sometime. Some of the scabs took longer to fall off and with
the new wool growing under them, they looked like an odd crop of mushrooms
(Plate II, fig. 2). The crop of wool after recovery was got analyzed, as regards
crimp, length, elasticity and tensile strength and was found to be normal. It,
therefore, appears, there is no permanent damage to the wool follicles.
There was no itching in the initial stages, but in later stages some animals were
seen scratching and responded to the scratch reflex.
The course varied from four to fifteen days. The appetite was maintained in
those cases in which there was no rigidity of the neck. There was a rise of tempera-
ture of one or two degrees in fatal cases, usually on alternate days.
PATHOLOGY
Post-mortem lesions
The skin was hard, congested, and about ten times thicker than the normal.
In some cases there were small depressions at the surface of the skin and in others
nodular eruptions, but in the majority of cases, however, the skin was like a sheet
of parchment. The cut surface, showed the presence of minute pustules (Plate II,
fig. 1). The blood vessels of the skin were extremely congested. No lesions, directly
referable to the main disease, were detected in any of the internal organs.
HISTOPATHOLOGY
The lesions are referable to a local infectious disease of the skin with the seat
of the germ in the epidermis. There is an inflammatory change of the cutis at the
epithelial margin, leading to rapid proliferation of the cells of the germinal layer with
cornification or parakeratosis and hyperplasia of the prickle cells (Plate III, figs. 1
and 2). The cells of the parakeratotic transitional layer preserve their nuclei which
are flattened and in section appear lentil-shaped. The cells of the superficial layers
are devoid of nuclei and stain poorly and uniformly. These parakeratotic cells, on
account of the pressure of the cornified tissue from above and the strong degree of
resistance of the wavy papillary body from below, are thrown into ridges and taps
and look under low power as a series of pillars composed of arched cells. There
are focal accumulations of polymorphonuclear leucocytes, usually at the base of the
arches at their junction with the prickle cells.
The blood vessels of the dermis are congested and those of the papillæ are
laden with a large number of leucocytes. In some cases the wool folicles are fewer
in number. The sebaceous glands are compressed and are not easily distinguishable
in grossly affected places.
The collogen fibres of the hypodermis are very thick and numerous as compared
with the normal and the blood vessels are congested and contain many polymorphs.
The causal organism is rod-shaped with infrequent branching. In sections
they appear of varying thickness. By gram weight they are Gram positive and
appear to stain uniformly but under higher power, most of these rods comprise of
rules of biscuit-shaped discs. These discs apparently stain more intensely along the
Indian J. Vet. Sci. Vol. XIX, Part I]
[PLATE I
FIG. 1. Wounds showing raw surfaces
FIG. 2. Showing hardening of the skin and wrinkles
PLATE II]
[Indian J. Vel. Sci. Vol. XIX, Part I
FIG. 1. Inside of the skin showing pitting and pus nodules
FIG. 2. A lamb four months after recovery showing lumpy wool
Indian J. Vet. Sci. Vol. XIX, Part I]
[PLATE III
FIG. 1. Hyper keratosis of the epidermis
FIG. 2. Mycelia in the epidermis
March, 1949]
H.
K.
LALL
AND
V.
R.
RAJAGOPALAN
3
margin and so appear sometimes as bipolar rods placed side by side in series. At
the surface of the skin, these rods break up into coccoid elements. The organisms
are distributed in largest numbers at or near the surface of the skin. They work
their way up along shaft of the wool follicles and sometimes reach as far down as the
papillæ of the hair. They are also found working into the epidermis in tortuous
courses along the poles of the sweat glands. Sometimes fragments are found dis-
tributed throughout the cornified tissues but they never apparently reach the prickle
cells or the dermis.
The peculiar distribution of the parasite suggests that they are normally present
on the surface of the skin and in the hot rainy months when there is a large outpour
of sweat, the stratum cornium becomes soft and affords the parasite a suitable site
for its growth and multiplication and the infection probably extends along the
sweat pores but the inflammatory changes induced at the stratum germinativum
and the attraction of leucocytes through the intercellular lymph spaces of the
prickle cells is a distant action possibly effected through the agency of a diffusible
toxin.
BACTERIOLOGY
The causal organism which was clearly a mycelium in morphology, could not be
cultivated as the lesions, in each case, had been previously treated with preparations
containing iodine. Detailed work on the bacteriology of the organism from other
outbreaks is under investigation and will be published on completion of the work in.
due course.
DISCUSSION
From the available literature it appears, that this condition has not been previ-
ously recorded in India. Seddon [1927-28] described a form of disease very similar
to what we observed except that in our cases the matting of the wool was not a
prominent feature. Bull [1922] isolated a mycelium from such cases which he named
Actinomyces dermatonmus. Morais [1922] described apparently the same disease
under the term 'Lumpy wool' and he associated the disease with an excess of mois-
ture owing to continued rainfall. Cases also were marked by 'Lumpy wool'.
Morais [1930] was unable to transmit the disease by contact or by scarification, but
Fethers [1939] was readily able to transmit the disease.
Among those who have described similar diseases in other species is Albiston
[1933] who reported the disease in calves. The organism isolated conformed to the
general description of A. dermatonomus except that colonies of his cultures were
greyish-white in contrast to the yellow colour typical of the type species. Stable-
forth [1937] and Edgar and Keast [1940] encountered similar conditions in the horse
caused by the same organism. The lesion started at the lip and spread to other
places. The disease has been reported in the goat by Beaton [1932] in which
lesions were found in the abdomen and the inner surface of the forelegs and thighs.
SUMMARY
A form of dermatitis in Hissardale lambs (Bikaneri-Merino crosses) caused by
a mvcelium is described along with the symptoms, lesions, and histopathology.
4
ACTINOMYCOTIC DERMATITIS IN CROSS MERINO LAMBS
[Vol. XIX, Part I,
REFERENCES
Albiston, H. E. (1933). Austr. Vet. J. 9, 107
Beaton, W. G. (1932). Annual Report of the Veterinary Deptt. Nigeria for 1932, 20.
[Cited by Hudson J. R. (1937).] Proc. Royal Soc. Med. 30, II, 1957
Bull, L. B. (1922). Aust. J. Exp. Biol. 6, 3, 301. Cited by Mason and Bekker (1934) vide infra
Edgar, G. and Keast, J. C. (1940). Austr. Vet. J. 16, 120
Fethers, Geo. (1932). The Pastoral Renew and Graz. Record 49, 156
Morais, I. P. (1922). Farming in South Africa 4, 452
Seddon, H. R. (1927-28). Vety. Res. Rep. No. 5, Deptt. of Agr., New South Wales, Australia
Stableforth, A. W. (1937). Proc. Roy. Soc. Med. 30, 2, 1455
STUDIES ON THE DRYING OF RINDERPEST VIRUS
By N. B. DAS, Indian Veterinary Research Institute, Mukteswar
(Received for publication on 6 August 1948)
T HIS paper describes experiments on the drying of rinderpest virus so as to
conserve its virulence. The undried virus does not retain the virulence for
very long in the Indian field conditions unless maintained on ice or otherwise suitably
refrigerated. The dried virus, on the other hand, can resist better the environmental
factors, chiefly climatic, which are responsible for the rapid deterioration of the
undried product.
Pfaff [1938] dried goat spleen vaccine in vacuo over calcium chloride and showed
that the dried vaccine was equal in its immunizing effect in cattle and buffaloes to the
untreated fresh spleen vaccine. Nair et al. [1942] also reported similar results.
Robles and Generoso [1934] prepared a vaccine by slow drying of spleen and lymph
glands of rinderpest infected animals over calcium chloride in the cold. We have
desiccated the virulent virus according to Pfaff as well as by other methods and have
also studied the stability of the dried product at such temperature ranges as would
be experienced in the transit of dried virus from the laboratory to the field.
EXPERIMENTAL
The virus contained in leucocytes as well as in spleen tissues has been desiccated
by different methods. The dried products were tested on hill bulls (Kumauni
cattle) for virulence at varying intervals after preparation to study the survival of
the virus under certain methods of drying and when exposed to different tempera-
tures for varying periods.
Experiments with leucocyte-cream
Blood of a hill bull, inoculated with rinderpest virus (E-line of this Institute)
was collected in sterile flasks containing sodium oxalate (0.2 per cent final concen-
tration). The oxalated blood was centrifuged to separate plasma and the white
cells were carefully collected with a pipette. The white cells were freed from the
contaminated R. B. C. by repeatedly centrifuging the cell suspension in normal saline.
One c.c. of this cream was kept at 4°C as control; 0.1 c.c. of this control sample
was tested on a hill bull along with the other samples described below.
Drying on filter paper
A piece of Whatman filter paper (No. 40; 2 sq. c.m.) was soaked with 1 c.c. of
the above leucocyte-cream and was dried in a vacuum desiccator over calcium
chloride, the desiccator being in an ice-salt mixture for the first few hours and then
at 0°C overnight. The evacuation was repeated a few times during the first five
hours in order to maintain a high vacuum. Ultimately the dried material was
ground in an agate mortar with 2 c.c. normal saline and the whole suspension was
injected subcutaneously into a hill bull (Table I).
5
B
6
STUDIES ON THE DRYING OF RINDERPEST VIRUS [ Vol. XIX, Part I
Drying by cold acetone
One c.c. of the leucocyte-cream was mixed with 5 c.c. of acetone (cooled in ice-
salt mixture for two hours prior to use) in a small test tube and was left in ice-salt
mixture for five minutes with occasional shaking. The whole precipitate was then
collected on a small filter paper (Whatman No. 40) in the refrigerator. The precipi-
tate, along with the filter paper, was dried in a vacuum desiccator over calcium
chloride as described above. The dried material was ground with 2 c.c. normal
saline and was inoculated into a hill bull (Table I).
Drying by cold alcohol
One c.c. of the leucocyte-cream was mixed as above with 5 c.c. of cold absolute
alcohol and was kept in ice-salt mixture for five minutes before filtration. The residue
on the filter paper (Whatman No. 40) was dried as above. The dried material was
similarly ground with 2 c.c. normal saline and was injected into a hill bull (Table I).
Arrangement of animals, inoculation and recording of temperature, etc.
All the experimental animals described here were kept in alternate stalls in
disinfected sheds in order to prevent spread of infection from one to the other.
Separate syringes were used for injecting different animals and the operator disin-
fected his hands after every injection. The control animals, inoculated with un-
treated virus, were isolated from the experimental animals by accommodating them
in a different shed and their temperature was always taken last of all. The thermo-
meters were sterilized by washing first with alcohol and then with carbolic acid
solution before recording the temperature of each animal.
TABLE I
Comparative rate of inactivation of leucocyte-virus dried by different methods
Method of drying
Fresh (untreated) . . . . . . . . .
Dried on filter paper . . . . . . . . .
Acetone dried . . . . . . . . .
Alcohol dried . . . . . . . . .
Reaction
Good
Good
Good
Delayed
Survival days
10
9
15
17
March, 1949 ]
N.
B.
DAS
7
From the above Table it appears that the leucocyte-cream virus was not com-
pletely destroyed either by drying on filter paper or by precipitation with cold
acetone. The delayed reaction in case of alcohol dried virus appears to be due to
the changed properties of the virus.
The above results prompted us to study if virulent rinderpest spleen virus could
be dried without loss of potency?
Experiments with spleen tissue
The spleen was collected from hill bulls on the 5th day of inoculation with the
E-line virus in a sterile vessel covered with ice and was minced twice in a cool and
sterile mincer. The resulting spleen pulp was dried by the methods described below.
Drying by cold acetone
The spleen pulp was agitated with five volumes of cold acetone (cooled in ice-salt
mixture) for three minutes and was filtered over Buchner funnel under suction. The
residue was treated again as described above and was finally kept in a sterile beaker
in vacuo over calcium chloride in order to get rid of the last traces of moisture, the
desiccator, covered with a black cloth, being in ice-salt mixture for two hours
and then at 0°C.
After 40 hours the dried tissue weighing 1.1 gm. (equivalent to 5 gm. fresh tissue)
was emulsified in 15 c.c. sterile broth and was inoculated subcutaneously into a
hill bull. The other half of the dried tissue was kept in a sealed tube at room tem-
perature and was tested after one month (Table II).
Drying by cold acetone and ether
The pulp was treated twice as described above with cold acetone and then with
twice its volume of cold ether on a Buchner funnel. The tissues which were almost
dry were finally kept, as above, over calcium chloride.
After 40 hours the dried tissue, equivalent to 5 gm. fresh tissue, was injected
into a hill bull as mentioned earlier and the other half was kept at room temperature
to be tested after a month's storage (Table II).
Drying by cold acetone in the presence of phosphate
The pulp was mixed with a solid phosphate mixture yielding pH 7.0 and was
treated with ten volumes of cold acetone for five minutes before filtration under
suction. The fine dry powder was then kept over calcium chloride as above.
After 40 hours the dried tissue, equivalent to 5 gm. fresh tissue, was injected into
a hill bull and the other half was stored at room temperature (Table II).
8
STUDIES ON THE DRYING OF RINDERPEST VIRUS [Vol. XIX, Part I,
TABLE II
Potency of virus dried by different methods and stored for one month at room temperature
(about 12°C)
Method of drying
Cold acetone
.
.
.
.
.
Cold acetone and ether
.
.
.
.
.
Mixed with phosphate and treated with acetone .
First test
Reaction
Good
Good
Good
Survival
days
15
10
10
Second test after
a month
Reaction
Good
Good
Moderate
Survival
days
11
13
13
From Table II it appears that spleen virus is not completely inactivated by
treatment with cold acetone under different conditions.
It was then proposed to determine the comparative rate of inactivation of the
virus dried by different methods as described above. Tables III, IV and V give
the figures when the virus is dried by cold acetone, cold acetone followed by ether
and cold acetone compared to that dried in vacuo over calcium chloride respectively.
At least two hill bulls were used for each test.
TABLE III
Comparative rate of inactivation of virus dried by cold acetone
Quantity of tissue inoculated. (Equivalent of fresh
tissue)
1.0 gm
.
.
.
.
.
.
.
.
0 01
gm
.
.
.
.
.
.
.
.
0.0001 gm . . . . . . . .
Dried tissue virus
Reaction
Good
Good
Delayed
Average
survival
days
11
14
21
Fresh tissue virus
Reaction
Good
Good
Good
Average
survival
days
12
11
10
Cold acetone dried virus appears to be active even in 0.01 gm. dose while in
0.0001 gm. dose the activity appears to be doubtful. The delayed reaction
followed by death may be due to the changed properties of the virus.
March, 1949.]
N.
B.
DAS
9
TABLE IV
Comparative rate of inactivation of virus dried by cold acetone and ether
Quantity of tissue inoculated. (Equivalent of fresh
tissue)
1.0 gm
.
.
.
.
.
.
.
.
0.01 gm. . . . . . . . .
Dried tissue virus
Reaction
Moderate
Delayed
Average
survival
days
15
17
Fresh tissue virus
Reaction
Good
Good
Average
survival
days
8
14
Cold acetone and ether appear to have inactivated the virus to a considerable
extent.
Drying in vacuo over calcium chloride
The pulp was uniformly spread over a sterile dish in a thin layer and was kept
in a vacuum desiccator with calcium chloride, the desiccator being in an ice-salt
mixture during the process of evacuation and then in the refrigerator.
The dried tissue was then ground to a fine powder and was emulsified in normal
saline immediately before inoculation (Tables V and VI).
TABLE V
Comparative rate of inactivation of virus dried with cold acetone and in vacuo over
calcium chloride
Quantity of tissue inoculated.
(Equivalent of fresh tissue)
1.0 gm . . . .
0.01 gm . . . .
Acetone dried
Reaction
Good
Delayed
Average
survival
days
9
15
Calcium chloride
dried
Reaction
Good
Good
Average
survival
days
10
12
Fresh tissue virus
Reaction
Good
Good
Average
survival
days
11
9
From Table V, calcium chloride dried virus ppears to be better than acetone
dried virus. This lot of acetone dried virus did not react in 0.01 gm. dose as described
in Table III.
Attempts were then made to determine the comparative rate of deterioration
on storage of the virus dried by cold acetone as well as in vacuo over calcium chloride
when inocculated in doses equivalent to 1.0 gm. fresh tissue (Table VI).
C
10
STUDIES ON THE DRYING OF RINDERPEST VIRUS [Vol. XIX, Part I
TABLE VI
Comparative rate of deterioration of dried virus at different temperatures
Temperature of storage
Freshly dried
.
.
. .
4°C . . . .
4°C . . . .
28°C. Being originally at 4°C for
4 months.
37°C
Room temperature (about 12°C) .
Period of storage
. .
2 months
4 months
7 days
15 days
3 months
Acetone dried
Reaction
Good
Good
Nil
..
..
Nil
Average
survival
days
9
11
No death
..
..
No death
Calcium chloride
dried
Reaction
Good
Good
Good
Good
Good
Nil
Average
survival
days
10
9
13
10
10
No death
Table VI shows that calcium chloride dried virus is superior to acetone dried
virus. Calcium chloride dried virus is active even after fifteen days storage at 37°C
and after four months storage at 4°C.
SUMMARY
Rinderpest virus contained in leucocytes can be dried by cold acetone as
well as on filter paper in vacuo over calcium chloride.
Experiments on the drying of rinderpest virus contained in spleen tissue
by cold acetone, cold acetone followed by ether, cold acetone in the presence of
phosphate as well as in vacuo over calcium chloride are described.
Virus dried in vacuo over calcium chloride appears to be the best and is
active up to 0.01 gm. dose while virus dried by cold acetone was sometimes found
inactive in that dose.
Virus dried over calcium chloride has been found potent after at least fifteen
days storage at 37°C and four months storage at 4°C. It is not inactivated on
storage for seven days at 28°C after being kept at 4°C for four months.
Acetone dried virus was found potent after two months storage at 4°C.
This work was carried out under the guidance of Mr J. R. Haddow.
REFERENCES
Nair, K. S., Krishnamurti, R. & Kalyanasundaram, G. S. (1942). Indian J. vet. Sci. 12,305.
Pfaff, G. (1938). Onderstepoort J. Vet. Sci. & Ani. Ind. 11, 263.
Robles, M. M. & Generoso, J. D (1934). Phillip. J. Ani. Ind. 1, 33.