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ORIGINAL ARTICLES
"ACTINOMYCOSIS"* AND ACTINOBACILLOSIS IN
ANIMALS IN INDIA †
BY
M. Y. MANGRULKAR, M. Sc., M.R.C.V.S. D.T.V.M. (EDIN.)
Assistant Pathologist, Imperial Veterinary Research Institute,
Mukteswar-Kumaun
(Received for publication on 4th May, 1938)
(With Plates XIX—XXIV.)
THE object of writing this paper is to call the attention of veterinarians
in India to the existence of " Actinomycotic " disease-conditions in animals,
and to emphasize the necessity of making a scientific diagnosis and allotting
the correct nomenclature to these conditions. For this purpose, a résumé
of the literature is also placed before them.
It appears that, in this country, the clinical side of the subject has been
sadly neglected, the few cases so far recorded indicating that the disease, which
is referred to broadly as " Actinomycosis ", was in most cases, discovered on
microscopic examination of material collected post mortem. A previous
clinical diagnosis of the disease has been suggested only in a few cases, while
in others, a chance finding of the actino-body has been instrumental in the dis-
covery of the " actinomycotic " character of the granuloma. It may be ar-
gued that the disease is of rare occurrence in India, but the records of this
Institute show that, contrary to the belief of veterinary research workers in
the past, typical cases of ' Lumpy Jaw ' and ' Wooden Tongue ' were not un-
common in North Bihar. Similarly, enquiries made, by the author, of stu-
dents who visit this Institute for training have elicited the information that
conditions resembling " Actinomycosis " in cattle do occur. It was suspected,
however, that the clinical diagnosis was usually not a scientific deduction and
that attempts to have such diagnosis confirmed microscopically were seldom
made. Under such circumstances, one need not wonder at the paucity of
material for cultural examination.
In this Institute, only fixed tissues have been received which have shown,
often unexpectedly, the presence of an " actinomycotic " disease. It will be
realised that in interpreting such granulomata, one has to go by the results of
work done in other countries without an opportunity to take up independent
research. It is highly probable that it is not the rarity of " actinomycosis "
* Although not in favour of the use of the non-specific comprehensive term "Actino-
mycosis ", the author has used it in this article for the sake of brevity and has indicated
such use by placing the term usually within inverted commas, as above.
† Presented at the 25th Session of the Indian Science Congress, Calcutta, 1938.
( 271 )
M. Y.
MANGRULKAR
277
In the collection of this Institute are specimens which have been received
from different parts of the country for microscopical examination and diag-
nosis. Although the material is only meagre it is sufficient to prove the exist-
ence of at least some of the actinophytoses in India. To add to the evidence,
and for comparative study, some material was especially obtained by Mr.
T. J. Hurley, M.R.C.V.S., D.V.S.M., I.V.S., (Offg. Veterinary Research Officer
incharge of Pathology, Imperial Veterinary Research Institute, Mukteswar)
from the collections of the Veterinary College, Madras. To Mr. Hurley and
to the authorities of the Madras College, I am greatly indebted for the help.
The following record of the result of examination of the various specimens is
not intended to give a histological study of the materials, for this has already
been done by many other workers [Bosworth, 1923 ; M'Fadyean, 1932]. It
is intended to show how with the help of available staining methods the etio-
logy of the actino-body can be ascertained with great success.
A. SPECIMEN 1357/1928
The material was received in May, 1928, in the form of two small pieces
of tissue removed from the tongue of a six-year old country-bred, emaciated
cow showing " swelling of the tongue with granulation tissue, difficulty in
taking food ; foaming saliva coming out from the mouth". The condition
was suspected to be " actinomycosis ".
The tissue on histological examination proved negative at first, but on
examination of sections prepared later " beautiful actinomyces clubs " were
demonstrated but the responsible organism was not determined. As the
material was exhausted, the author had to be satisfied with re-examination
of available stained sections. Fragments of an actino-body, surrounded
by a zone of cellular reaction, were seen in each section. The entire lesion was
microscopic, non-elevated and situated superficially, embedded in the marked-
ly hypertrophied epithelial layer of the tongue (Plate XIX fig. I). The
focus had an extremely ill-formed outer capsule.
On restaining some of the preparations, not Gram-positive but Gram-
negative small bacilli were detected, the lesion resembling the one described
later (Specimen 327A/1936, p. 283).
This case appears to be the first in the records of this Institute, which
from the clinical and histological features can be interpreted as one of actino¬
bacillosis.
B. SPECIMEN 1557/1929
The information available in connection with this specimen is rather limi-
ted. The tissue was removed from the shoulder of an ox and carcinoma was
suspected.
A diagnosis of " actinomycosis " was given in 1929. In freshly cut sec-
tions (1937), the tissue showed itself to be the product of connective-tissue
reaction in the subcutis. In it were found microscopic pus-foci which faded
out peripherally into the surrounding granulomatous tissue. The centre of
C
278
"Actinomycosis"
and Actinobacillosis in Animals
several foci revealed actino-bodies with clubs which were larger and more
prominent than in actinomycosis (Plate XIX, fig. 2). In suitable sections, it
was discovered that the minute central portion of the ray-colonies was occupied
by a cluster of Gram-positive bacilli. Their exact identity could not be deter-
mined but such characters, as are usually associated with diphtheroids, were
observed, e.g., uneven staining, filamentous and coccoid forms, and club-like
swellings. Fortunately a preparation of Corynebacterium pyogenes in tissue was
available for comparison which afforded strong suggestive evidence in favour
of the view that the bacteria in the actino-bodies were diphtheroids.
In and around the pus-foci eosinophiles were very frequently seen but a
careful search for worms or worm-ova, in sections and in caustic potash prepa-
rations, proved entirely negative. No foreign body giant-cells were seen.
Of the skin, only the deeper layers of the epidermis were visible in certain
sections but no carcinoma was detected.
The case was evidently one of actinophytosis due to a pleomorphic Gram-
positive bacillus (? Corynebacterium) but not due to a Streptothrix.
C. SPECIMEN 1857/1931
The material, which unfortunately could not be traced, was received from
the Principal, Madras Veterinary College. It belonged evidently to the
buffalo, referred to in the Annual Report of the Civil Veterinary Department,
Madras, 1927-1928. It may be recalled that a Gram-positive Streptothrix
was observed in sections. Although all necessary information is not avail-
able with regard to this material, it is possible that the case was one of actino-
mycosis.
D. SPECIMEN 1862/1931
This specimen forms the subject of the paper presented by Datta [1933]
to the Indian Science Congress in 1933. The material was received from the
Superintendent, Sind and Rajputana, and consisted of " actinomycosis tu-
mours removed from both parotid glands of a
buffalo heifer ".
The tumours were situated " about three inches below the base of the ears "
and " measured three inches in diameter ". They were first noted as walnut-
sized growths when the animal was about a year old and had grown to the size
mentioned in six months at the end of which period they had begun to " emit
a discharge " (fistula-formation) of the nature of stringy " thick inspissated
pus ". There was difficulty in deglutition and breathing. After removal of
the tumours, iodine treatment was adopted and " the heifer showed marked
improvement ".
On first examination of the tumour in October, 1931, " actinomycosis "
was confirmed except that no Gram-positive filaments were seen. Subse-
quent examination of sections made in August, 1932, and stained by Gram¬
Weigert revealed mycelial filaments inter-twined inside the " ray-fungus ".
Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Pt. IV ]
PLATE XIX
FIG. 1.
Actinobacillosis of bovine tongue
(1357×1928; ×36)
FIG. 2.
Actinophytosis of bovine shoulder
(1557×1921) ; × 396)
FIG. 3.
Actinobacillosis of parotid gland
(1862×1931 ; ×54)
FIG. 4.
Actinobacillosis of parotid gland
(1862×1931 ; ×792)
PLATE XX
[ Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Pt. IV.
FIG. 1.
Hill bull 749, showing Actinomycosis of the head
FIG. 2.
Colonies of Actinomyces in cheek nodule
(175/1934; ×81)
FIG. 3.
Actinomycotic lesion in the skin (175/1934 ; ×40)
M. Y.
MANGRULKAR
279
A re-examination of the material was undertaken this year (1937) and
numerous sections cut and stained by different methods. Histologically,
the tissue represented an advanced chronic lymphadenitis with formation of
abundant connective tissue wherein several small actino-bodies, usually in
groups, were seen (Plate XIX, fig. 3). These were surrounded by a varying
amount of cell-detritus. By Gram-Weigert method of staining, no evidence of
a Gram-positive organism was detected. By modified Ziehl-Neelsen method
(using two per cent sulphuric acid instead of twenty-five per cent), however,
a beautiful and instructive picture was obtained (Plate XIX, fig. 4). The peri-
pheral clubs were well formed and numerous. No filaments of streptothrix
were observed in the colonies. Instead, centrally to the zone of clubs, and
rarely in the peripheral zone itself, numerous masses of (Gram-negative)
short bacilli were seen which could be clearly made out in the smaller (? early)
single colonies.
The results of the last examination agree with those of 1931 but cannot
be reconciled with the observations recorded in 1932. The possibility of an
actinomycetic infection is so remote as to be ruled out in view of the fact that
actinomycosis does not spread by the lymphatics. I may here quote from
Cope [1930] :—
" Concerning the conduct of the actinomycotic fungus in the tis-
sues of the body, there are three points which are peculiar
and characteristic. The first is the way in which the pro-
cess advances almost always by contiguity of the tissue. Blood
transmission occasionally occurs, as for example infection of
the liver from the appendix through the portal veins, but
usually the slow inflammation progresses by contiguity of tissue,
often receding in one place and advancing in another. The second
characteristic is the curious immunity of the lymph glands to the in-
fection. I have never seen a lymphatic gland infected by Actinomy-
cosis, and have never seen any authentic record of such a happening.
This feature places it in sharp contrast not only with the actinobaci¬
llosis infection of cattle, but also with almost every other variety of
infection both acute and chronic ".
The clinical history and histological features including absence of Gram-
positive filaments in this case are thus indicative of a certain actinobacillary
infection. It is unfortunate that no information about the existence of disease
in other organs was available.
E. SPECIMEN 175/1934
This material was collected locally from a clinical case in a Hill-Bull (No.
from 749) while it was alive (Plate XX, fig. 1) and consisted of a nodular
growth the right cheek, and a piece of skin. " Typical lesions of Actino " was
the diagnosis given. Bone-lesion was suspected during life.
In this case it was unfortunate that no other observations were recorded.
The animal was subsequently destroyed but no post mortem examination was
conducted. The site of the removed skin was similarly not recorded, but was
probably in the neighbourhood of the nodule.
280
"Actinomycosis" and Actinobacillosis in Animals
On histological examination there could be no doubt about the actino-
mycotic nature of the lesions. In the cheek, the " nodule " consisted of an
abscess, about 0.75 cm. in diameter, in the thickness of the cheek away from
the outer surface. The focus consisted mainly of polymorphs and contained
numerous actino-bodies in different stages of formation. The Gram-positve
branching streptothrix could be easily demonstrated in the lesion (Plate XX,
fig. 2). It showed occasional terminal swellings, and slightly uneven Gram-
positive character.
In the skin the dermis showed signs of connective tissue proliferation
(Plate XX, fig. 3) and contained a few pus-foci with actino-bodies due to the
Gram-positive streptothrix.
F. SPECIMEN 245, B/1934
The material was from a twelve-months old, cocoanut-sized, inoperable
tumour of the lower lip of a bullock from Sind. Actinomycosis was suspected
and confirmed by the evidence of " a few actino-granules " and " numerous
mycelial masses ".
It was unfortunate that old sections of this material were not available
and that the material had become exhausted except for pieces of what appeared
to be the reacting well-formed connective tissue. Repeated attempts to verify
the diagnosis by making fresh sections proved of little value as nothing but the
old granulomatous tissue was encountered in sections.
Yet, by the situation of the lesion [Cf. Specimens 175/1934 (p. 279) and
194/1937 (p. 284)] and by the diagnosis already given in 1934, there seems
little doubt that the case was one of actinomycosis.
In connection with the formation of abundant connective tissue in this
case, the quotation from Cope [1930] may be continued :
"
Thirdly, everyone who has seen much of actinomycosis must
have pondered over the remarkable reaction which the fungus causes
in the connective tissues. I think it was Unna who pointed
out that here we have an almost unique example of tissue reaction at
a distance, for the changes may take place at a considerable distance
from the place where the fungus is situated, and it is possible to cut
many sections of the curiously hard gristly fibrous tissue, without find-
ing any filaments of the parasite. This connective tissue reaction
differs considerably from that found in tuberculosis and syphilis not
only in the relative proportion of the various kinds of cells found in it,
but also in the fact that the blood vessels are not obliterated. Exact¬
ly how the fungus causes the reaction is unknown ".
G. SPECIMEN 192/1936
This specimen consisted of a piece of tumour from the nostril of a buffalo
from the Civil Veterinary Department, Agra, U. P., presumably suspected for
Schistosomic nasal granuloma. No other information was available in this
case. A preliminary diagnosis of " actinomycosis " was given, but as no Gram-
positive micro-organism was seen in sections a detailed study of the specimen
was continued. At the end, the growth was diagnosed as actinobacillosis.
PLATE XXI
[ Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Part IV
FIG. 1.
Actinobacillary nasal growth (192/1936 ; ×67)
FIG. 2.
Actinobacillary nasal growth
(192/1936; ×720)
FIG. 3.
Villous growths on the heart
(296/1936 ; × 18)
Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Part IV ]
PLATE XXII
FIG. 1.
Villous growths on the pericardium (296/1936 ; × 32)
FIG. 2.
Pericardium. Streptothrix colony
(296 1936 ; × 50)
FIG. 3.
Pericardium. Streptothrix colony
(296/1936; × 211)
FIG. 4.
Pericardium, showing Streptothrix colony and
worm-eggs (296/1936 ; × 50)
M. Y.
MANGRULKAR
281
The tumour consisted of a highly cellular granulomatous tissue under the
ciliated nasal mucous membrane with numerous foci of cellular reaction,
containing polymorphs and showing occasional giant-cell formations. In
these foci were seen, as a rule, groups of small, often apparently confluent
actino-bodies (Plate XXI, fig. 1) with well-formed clubs. Again, no Gram-
positive mycelium was seen, but by modified Ziehl-Neelsen method of staining
masses of (Gram-negative) bacilli (Plate XXI, fig. 2) were detected amongst or
just under the clubs except in the smallest (? early) colonies, where they were
situated in the centre of the actino-body. The centre of the large (? older)
colonies was left as an unstained more or less homogeneous mass.
This specimen with which Specimen 1862/1931 (p. 278) affords a very re-
markable comparison, offers good material for the study of typical actinobaci¬
llosis. Enquiries made for obtaining more information and material, especially
for cultural examination, were unfortunately disappointing, the animal having
died during the interim.
H. SPECIMEN 296/1936
This material, the only one of its kind in the collection of the Institute,
was received from Mr. Mutappa, Veterinary Assistant Surgeon, Ponampet,
Mercara, to whom I am indebted for also furnishing a complete history of the
case. The patient was a cross-bred spaniel of about two years which after a
course of treatment for suspected distemper came under the observations of
Mr. Mutappa on the 6th October 1936. High fever, dyspnoea, and an anxious
look were noticed. The animal usually stood with the elbows turned out.
The pulse was weak and irregular. Daily injections of Omnadin did little
good and on the third day the animal was much worse, with laborious breath-
ing and an almost inaudible heart-action. The patient died on the 9th
October 1936. On autospy the " thoracic cavity was found to be filled with
serosanguineous fluid. The pericardial sac was also highly enlarged with the
sero-sanguineous fluid. Round about the pericardium, there were fibrinous
growths. The lungs were normal. No parasites either in the lungs or the
intestines were detected by me".
The material submitted for examination was heart muscle and pericar-
dium in formalin.
With the naked eye, nothing beyond a fine villous proliferation on the
outer surface of the heart (epicardium) and on the inner surface of the pericar-
dial sac could be seen. On histological examination the suspicion of a neo-
plastic disease was at once proven to be mistaken. The fine velvety appear-
ance of the tissue was revealed to be a highly vascular granulomatous reaction
(Plate XXI, fig. 3, and Plate XXII, fig. 1).
In the epicardial lesion no actino-bodies were seen but fragments of a
Gram-positive micro-organism in the form of fine branching filaments and
coccoid forms, some contained within wandering cells, were observed,
Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Part IV ]
PLATE XXIII
FIG. 1.
Pericardium showing worm-eggs
(296/1936 ; × 50)
FIG. 2.
Actinobacillosis of bovine tongue
(327-A/1936; × 36)
FIG. 3.
Actinomycosis of gum of cow
(191/1937; × 81)
PLATE XXIV
[ Ind. Jour. Vety. Sci. & Anim. Husb., Vol. VIII, Part IV.
FIG. 1.
Actinomycosis of gum of cow
(194/1937; × l35)
FIG. 2.
Actinophytosis of bovine udder
(194 1937 ; × 81)
FIG. 3.
Actinophytosis of bovine udder
(194 1937 ; × 576)