NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
RICKETTSIA OF DOMESTICATED ANIMALS
(A SHORT REVIEW)
By S. L. MANJREKAR, G.B.V.C., ASSOC. I.D.R.I., PH.D. (BACT.), Assistant
Disease Investigation Officer (Sheep & Goats), Bombay State, Poona
(Received for publication on 7 October 1952)
(With Plates IX-X)
R ICKETTSIA is the generic name given to certain minute, bacteria-like
bodies which are found in the alimentary canal of insects and other arthropods.
They are frequently associated with different forms of typhus or exanthematous
fevers in man.
From the very start rickettsiosis has been regarded as an infectious condition
involving the blood and producing a purpuric fever. Rickettsia, as infective agent
producing spotted fever in man, was first recorded in 1909 (Ricketts) and since then
number of workers—Ricketts and Wilder, Edm. Sergent, H. Foley and Viallette,
da Rocha Lima—are concerned in confirming the above findings.
Rickettsia of veterinary importance
Cowdry [1925] first described Rickettsia ruminantium as the causative agent
of 'heartwater' disease of sheep and cattle, which prior to his discovery was considered
to be caused by an ultra-microscopic virus. Later on various research workers
from different parts of the world have ascribed some species of Rickettsia causing
local as well as systemic disease conditions in domesticated animals.
CLASSIFICATION OF RICKETTSIA
Twelve species of pathogenic Rickettsia of veterinary importance have so
far been reported as causing diseases in domesticated animals [Lestoquard and
Donatien]. In India only R. conjunctivae, R. canis and R. belgaumii have been
found to be causing any disease condition so far. Systematic classification of
Rickettsia was till recently a debatable question, but now, scientists working on
this problem, have classified them tentatively in the family Rickettsiaceae [Pinkerton,
1936].
In the veterinary literature, Lestoquard and Donatien have broadly divided
Rickettsia into two classes, viz. those producing local affections while others producing
systemic diseases.
Rickettsia causing local affections
Under this, are included all those Rickettsia causing conjunctivitis in animals,
viz. (a) R. conjunctivae, (b) R. conjunctivae bovis.
Coles [1931-1936] showed that conjunctivitis of sheep, goats and cattle was
associated with a Rickettsia, in South Africa. His observations have since been
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RICKETTSIA OF DOMESTICATED ANIMALS [ Vol. XXIV, Part IV,
confirmed in North Africa by Donatien [1937] and by Shirlaw and Shankarnarayan
[1947] in India. The condition is associated with acute pain, lacrimation and
photophobia. Strangely enough the condition is restricted to one eye only. Corneal
ulceration has been recorded in a few cases. Clinically too the disease is recorded in
a few cases and in Africa it was found to be similar in many respects to the eye
condition reported in India by Shirlaw and Shankarnarayan. Similar condition
of eye infection amongst cattle and sheep is also recorded in northern parts of Queens-
land but its aetiology is not yet known.
Rickettsia producing systemic diseases
Donation and Lestoquard have again sub-divided these into two groups.
(a) Rickettsia of reticulo-endothelial system.
(b) Rickettsia of monocytes.
(a) Rickettsia of reticulo-endothelial system. Under this sub-head the following
are described.
(i) R. ruminantium, (ii) R. avium.
(i) R. ruminantium [Cowdry, 1925]. 'Heartwater' disease of sheep was shown
to be due to a Rickettsia but it became very difficult to demonstrate the organisms
in the tissues, until Jackson discovered that the scrapings of internal walls of the
larger veins often showed these parasites. The incubation period is nine days.
There is fever and allied symptoms which usually end fatally. Recovered animals
are immune in the sense of carrier immunity, but may act as reservoirs of these
organisms.
On post-mortem one may find hydropericardium, hydrothorax and ascites with
haemorrhages on the endocardium, congestion and oedema of lungs and enlarged
spleen.
The disease is transmitted by the tick Amblyomma hebraeum in which the infec-
tion occurs during larval and the nymphal stages, the disease is transmitted in the
subsequent stage but the Rickettsia does not pass through the egg.
(ii) R. avium [Carpano, 1936]. This micro-organism was described in pigeons
in Egypt, but till now nothing more is known about it.
(b) Rickettsia of monocytes. The following Rickettsia have been classed under
this sub-head:
(i) R. canis
(ii) R. ovina
(iii) R. bovis
(iv) R. belgaumii
(i) R. canis [Donatien and Lestoquard, 1935]. During investigations on the
infection of Piroplasmacanis and Lishmania of dogs in Algeria, this parasite was
encountered and the common dog tick Rhipicephalus sanguineus was incriminated
as the transmitter of the infection. Mudaliar [1944] has recorded R. canis from
a dog showing febrile illness.
Indian J. vet. Sci., Vol. XXIV, Part IV ]
PLATE
IX
R. ruminantium
R. canis
R. ovina × 15.10
R. bovis
R. conjuctivae
PLATE X
[Indian J. vet. Sci., Vol. XXIV, Part IV
Unnamed Rickettsia
R.
belgaumii
R. belgaumii
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RICKETTSIA OF DOMESTICATED ANIMALS [ Vol. XXIV, Part IV,
behaviour quite different from the latter or rather from all the Rickettsiae of domesti-
cated animals. In all these respects it is nearer to R. mooseri of man causing Murine
typhus to which it has been found to be akin on cross immunity tests.
It produces a mild systemic disease amongst sheep and goats characterised
by sudden onset and rise in body temperature by about 3° to 4° F. above the normal.
Fever is of a remittent or intermittent type with no mortality. The vector is found
to be a tick, Haemophysalis bispinosa.
R. belgaumii has been successfully transmitted to healthy guineapigs, which
show a definite thermal and scrotal reaction after an incubation period of three
to nine days. The strain can be easily maintained in white rats for a long time
(roughly 300 days). In this respect it differs from other Rickettsiae of domesticated
animals, since they are not transmissible to the laboratory animals.
That R. belgaumii is a new species has been concluded on the following grounds :
(1) All the Rickettsiae isolated from domesticated animals are not transmissible
to guineapigs—whereas the one isolated by Manjrekar was transmissible to the
male guineapig with a definite thermal and scrotal reaction resembling the Neill-
Mooser's reaction in Murine typhus.
(2) It is intracytoplasmic in the monocytes of the blood of sheep and goats.
(3) Rickettsia isolated was transmissible to white rats and could be kept up to
300 days.
(4) It could be retransmitted to sheep and goats with a definite thermal and
other allied syndrome of symptoms.
(5) It could be recovered and demonstrated under microscope in the peritoneal
and scrotal scrapings of the guineapigs.
(6) (a) Ticks which were found to be carriers were—Haemophysalis bispinosa.
(b) The disease could be reproduced in sheep and goats from these ticks.
(7) Serological findings did not show any agglutination in titres over 1 in 500
against OX19 whereas in Murine typhus it shows agglutination even at higher
dilutions as 1 in 2000 or so.
(8) Neutralization test proved that the strain of Rickettsia belgaumii was different
from that of Murine typhus Rickettsia as the serum of recovered sheep and goats
did not neutralize the Murine typhus Rickettsia whereas R. belgaumii was neutralized
R. phagocytophilia [Angus Fogie, 1951].—This organism, which has been seen
in almost all the leucocytes of the circulating blood of the sheep suffering from
a febrile illness in Scotland, is also shown to be transmitted through ticks.
It is seen in neutrophiles, basophiles, eosinophiles, lymphocytes and the mono
cytes. It causes a mild febrile illness in sheep and goats. Artificially it has been
found to be pathogenic to cattle but not to laboratory animals.
R. suis: Cause of a disease like swine fever.
R. burneti: Cause of 'Q' fever.
Un-named Rickettsiae
(a) Hudson [1950] has definitely established the existence of a new but mild
febrile condition amongst cattle in England, caused by a Rickettsia transmitted
December, 1954 ]
S.
L.
MANJREKAR
221
through ticks. He also detected those parasites in the monocytes of circulating
blood of naturally affected and experimentally infected animals. All these Rickettsiae
are not transmissible to laboratory animals such as guineapigs, etc.
(b) Pellisier, Troquereau and Trinquier [1950] have described a condition in
cattle which they ascribe to a Rickettsia of the exantheramic typhus group.
(c) A member of the same group—the cause of a condition in dog like that of
the exantheramic typhus group has been isolated, for which no special name has
been suggested.
SUMMARY
Rickettsia of veterinary importance have been reviewed in brief. Twelve
species have been described together with the conditions they cause in domesticated
animals.
ACKNOWLEDGEMENT
I feel extremely obliged to Dr J. Richard Hudson and Editor, The British
Veterinary Journal, for allowing me to publish a photo from one of the issues of
The British Veterinary Journal.
I am also very much thankful to the Director, Pasteur Institute, Algerie, for
so kindly permitting to make use of the photographs from their journals.
REFERENCES
Angus Fogie. (1951). Studies on the infectious agent of tick-borne fever in sheep. J. Path. Bact.
63, 1-15
Coles, J. D. W. A. (1931). A Rickettsia-like organism in the conjunctivitis of sheep. Rep. of Dir.
Vet. Ser., Onderstepoort, 17, 175
Carpano M. (1936). L'infezione da Rickettsiae negli uccelli (Rickettsia avium). Profilassi, fasc. 1.
Ann. IX, 1
Cowdry E. V. (1926). Rickettsiae and disease. Arch. Path. Lab. Med. II, 59
Donatien A. and Lestoquard F. (1935). Existence en Algerie d'une Rickettsia due chien. Bull. Soc.
Path. exot, t. 28, juin
Donatien A. and Lestoquard F. (1935). Sur la persistance de Rickettsia canis dans l'organisme du chien,
apres guerison. Bull. Acad. Med., t. 114, 857
Donatien A. and Lestoquard F. (1936). Recherches sur Rickettsia canis, Comparasion arec Rickettsia
conori. Ibid. 99 decembre
Donatien A. and Lestoquard F. (1937). Existence de Rickettsia conjonctivae due mouton en Algeric.
Bull. Soc. Path. exot., t. 30
Donatien A. and Lestoquard F. (1936). Rickettsia boris, nouvelle espece pathogene pour le boeuf. Ibid
Hudson J. R. (1950). The recognition of tick-borne fever as a disease of cattle. Brit vet. J. 106,
1, 3-17
Lestoquard F. and Donatien A. (1936). Sur une nouvelle Rickettsia du mouton. Bull. Soc. Path
exot. t. 29 novembre
Maujrekar S. L. (1949-51). Rickettsiosis in sheep and goats in the State of Bombay. (Thesis sub-
mitted to the University of Poona)
Mudaliar S. V. (1944). Canine rickettsioses in South India,—A preliminary note I.V.J., 20, 163-164
Pellisier, Troquereau and Trinquier. (1950). Bull. Soc. Path. ent. 43, 168-176
104-AR/54.
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RICKETTSIA OF DOMESTICATED ANIMALS
[ Vol. XXIV, Part IV
Pinkerton H. (1936). Criteria for the accurate classification of the rickettsial diseases (Rickettsioses) and
of their etiological agents. (As cited by John C. Snyder, in Viral and Rickettsial Infections of
Man, edited by Thomas M. Rivers, Lippincott Publication)
Von Prowazek. (1907, 1914). (Cited by Philips Thygeson, and John C. Snyder, in the Viral and Rickett-
sial Infections of Man, edited by Thomas M. Rivers, Lippincott Publication)
Ray H. N. (1950). Rickettsiosis in Indian sheep. Sci. and Cult. 15, 284-285
Ricketts H. T. (1909). A micro-organism which apparently has a specific relationship to Rocky Moun-
tain spotted fever. A preliminary report. J. Amer. med. Ass. 52, 379-380
Ricketts H. T. and Wilder R. M. (1910). The relation of typhus fever (tabardillo) to Rocky Mountain
spotted fever. Arch. int. Med., 5, 361-370
Ricketts H. T. and Wilder R. M. (1910a). The transmission of the typhus fever of Mexico (tabardillo)
by means of the louse (Pediculus vestamenti). J. Amer. med. Ass., 54, 1304-1307
Ricketts H. T. and Wilder R. M. (1910b). The etiology of the typhus fever (tabardillo) of Mexico City.
A further preliminary report. J. Amer med. Ass., 54, 1373-1375
da Rocha Lima H. (1912, 1916, 1919, 1930). (Cited by Max Theiler and John C. Snyder, in the Viral
and Rickettsial infection of man, edited by Thomas M. Rivers, Lippincott Publication)
Edm. Sergent H. Foley. C. Villatte. (1914). Sur des formes microbiennes abondantes dans le corps
des Poux infectes par le typhus exanthematique, et toujours absentes dans les Poux temoins non
infectes. C. R. Soc. Biol. t. LXXVII 19, juin, 101
Shirlaw J. F. and Sankarnarayan N. S. (1947). Conjunctivitis of sheep and goats in India. Indian.
J. vet. Sci. 17, 69-73
GLYCOGEN AND ITS PROBABLE SIGNIFICANCE IN
EIMERIA TENELLA RAILLIET AND LUCET, 1891
By B. S. GILL and H. N. RAY,* Indian Veterinary Research. Institute,
Mukteswar, Kumaon, U. P.
(Received for publication on 29 July 1954)
(With Plate XI)
G LYCOGEN and paraglycogen are the most widely distributed carbohydrates
in parasites, the latter so far has, however, been reported only from protozoa
[von Brand, 1952].
Giovannola [1934] found glycogen occurring in insignificant amount in
sporozoites within the oocysts; in schizonts, microgametocytes and young macro-
gametocytes of Eimeria falciformis and E. steidae; its concentration rising high
in older macrogametes and oocysts. Lillie [1947] demonstrated as pinkish matrix
of glycogen in E. steidae occurring in hepatic cells and lying in bile ducts.
Edgar, et al., [1944] employing iodine staining technique demonstrated some
glycogen lying centrally in sporozoites of E. tenella, also in macrogametes which
on growth increasingly accumulated larger quantities of it. Unsporulated oocysts
were full of glycogen which was exhausted on sporulation but they failed to demon-
strate the polysaccharide from schizonts, merozoites, microgametocytes and micro-
gametes.
MATERIAL AND METHODS
White Leghorn chicks, 1-2 weeks old, reared coccidia-free were employed in
these studies. The infective brew of oocysts was raised by feeding a single typical
oocyst of E. tenella [Barber's technique, Barber, 1914 ; Chambers, 1923]. Oocysts
were sporulated and preserved in 2.5 per cent solution (aqueous), of potassium
dichromate. Chicks were heavily infected per os. They were destroyed at four-hour
intervals, from 24 hours to eighth day (inclusive) of infection. Their caeca were
harvested with minimum loss of time after death (within a couple of minutes),
to avoid intracellular degeneration of the polysaccharide. Two to three mm. long
pieces of caeca were promptly fixed in alcohol-formalin mixture (9 : 1) for 24 hours
and the procedure as detailed by Glick [1949] was adopted to prepare the material
and staining the sections with Bauer-Feulgen technique (after Bensley); which
depends upon hydrolysing the polysaccharide into aldehyde, the latter group being
visualised as reddish-violet on. treatment with leucobasic-fuchsin. At each stage of
infection portion of caecum was longitudinally split open, mucosa scraped and this
material smeared over clean glass slides, which were immediately dropped in
Coplin jar containing alcohol-formalin fixative (10-15 minutes), washed in 95 per
cent alcohol and by running through descending grades of alcohol brought down
to water. They were, subsequently, treated in the same way as the sections. This
* At present Professor of Protozoology, School of Tropical Medicine, Calcutta.
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Indian J. vet. Sci., Vol. XXIV, Part IV ]
PLATE
XI
(Photomicrographs showing reactions of Eimeria tenella for various polysaccharides. Coloured
filters have been used, wherever necessary to give the proper photographic effect. Fig. 2 has been
slightly touched to accentuate the reactive zone but nothing has been added. Fig. 3, 11 and 12 are
taken from smear preparations and the rest are from sections. Hours within parentheses denote the
stage of infection.)
Various staining techniques used in the preparations photographed for preparation of this plate
are as under:—
Fig. 1-4: Bauer—Feulgen technique for glycogen.
Fig. 5 : Method of Hotchkiss, and
Fig. 6 & 7 : Method of Jarvi and Levanto for polysaccharides in general.
FIG.1.
Gametocytes (120 hrs. × 1600). Note the considerable glycogen reserve in the parasite.
FIG. 2. Smaller second generation merozoites (120 hrs. × 1400). Note the presence of glycogen
on both sides of the nucleus, the latter appearing as ' unstained' area.
FIG. 3. Second generation merozoites (120 hrs. × 1600). Note ante-nuclear glycogen reserve.
FIG. 4. Mature microgametocyte and macrogametes (140 hrs. × 1200). Inset is an oocyst.
Note intense reaction at this endogenous stage.
FIG. 5. Gametocytes (148 hrs. × 900). Note positive reaction for polysaccharide in the peri-
pheral globules.
FIG. 6. Second generation schizonts (8 hrs. × 900). Note intense reaction for polysaccharide
both in the cytoplasm and karyosomes.
FIG. 7. Second generation merozoites (120 hrs. × 2250). Showing positive reaction for poly-
saccharide in the karyosome.
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GLYCOGEN IN EIMERIA TENELLA
[ Vol. XXIV, Part IV
Evidence has been presented that the parasite metabolizes its glycogen from
host's blood sugar, and this probably occurs though the activity of the enzyme-
alkaline phosphatase.
By employing the technique of Hotchkiss [1948] after saliva digestion, it was
deduced that glycogen is the only detectable carbohydrate present in this coccidium.
ACKNOWLEDGEMENT
The authors are indebted to the Indian Council of Agricultural Research for
the award of Research Fellowship (Parasitology) to one of as (B.S.G.) which made
this investigation possible; and also to the Director, Indian Veterinary Research
Institute, for providing the laboratory facilities.
REFERENCES
Armer, J. M. (1944). J. Parasit., 30 : 131
Barber, M. A. (1914). Philipp. J. Sci; 9 : 307
Blaschko, and Jacobson, W. (1951.) Cytology and Cell Physiology, Published by the Clarendon Press,
Oxford, pp. 358
von Brand, T. (1932). Z. Parasitenk; 4 : 753
—
(1952). Chemical Physiology of Endoparasitic Animals. A. P. Inc. Publishers, N. Y.
Buetschli, O. (1885). Z. Biol., 21: 603
Chambers, R. (1923). J. Bact., 8 : 1
Edgar, S. A., Herrick, C. A. and Fraser, L. A. (1944). Trans. Amer. micro. Soc., 15 : 435-757
Fauré-Fremiét, E. (1913). Arch. Anat. micro., 15: 435
Giovannola, A. (1934). Arch. Protistenk., 83: 155
Glick, D. (1949). Techniques of Histo-and Cytochemistry. A. P. Inc., N. Y.
Hale, A. J. (1953). Quart. J. micr. Sci., 94 : 303.
Hotchkiss, R. D. (1948). Arch. Biochem., 16: 131
Jarvi, O. and Levanto. (1950). Acta path. microb. Scand., 27 : 473
Lillie, R. D. (1947). J. Lab. Clin. Med. 30 : 76
Maupas, E., (1886). C. R. Soc. Biol., 88: 342
Morita, Y. (1938). J. Ocient. Med., 28: 3 (English summary)
Pratt, I. (1940). Trans. Amer. micr. Soc., 59: 31
Ray, H. N. and Gill, B. S. (1954). Ann. Trop. med. & Parasit., 48, 8
Szwejkowska, G. (1929). Bull. into. Acad. Yougoslave Sci. B. 1928 : 489
Trier, H. J. (1926). Z. vergl. Physiol. 4 : 305
Usuelli, F. (1930). Wiss. Arch., Landw. B., 3: 4
Waxler, S. H. (1941). Amer. J. Hyg., 134 : 19
Westphal, A. (1934). Z. Parasitenk ; 7 : 71
Wislocki, G. B. and Dempsey, E. W. (1946). Anat. Rec., 90 : 219
ON THE OCCURRENCE OF MUCOPOLYSACCHARIDES
IN EIMERIA TENELLA RAILLIET AND LUCET, 1891
By B. S. GILL and H. N. RAY,* Indian Veterinary Research Institute,
Mukteswar, Kumaon, U.P.
(Received for publication on 29 July 1954)
(With Plate XII)
WHILE
investigations on the occurrence of nucleic acids in E. tenella were
being conducted (still unpublished) certain stages of the parasite displayed
intense metachromasy with toluidine blue. This led the authors to investigate
the occurrence of mucopolysaccharide in this coccidum by histo-chemical methods.
PROCEDURE
One to two weeks old White Leghorn chicks reared coccidia-free, were heavily
infected per os with a pure line brew of E. tenella. Chicks were destroyed at 4-hour
intervals from 1-8 days after infection. Small pieces (2-3 mm.) of caeca were fixed
in Carnoy's, Serra's, and Helly's fluids. Favourable material from caecal mucosa
was smeared on clean microslides and fixed in fluids mentioned above. Techniques
of Lison [1935] after Sylven [1947] and Brachet [1953] ; and of Hale [1946] were
adopted for demonstrating mucoid sulphate and hyaluronic acid type polysaccharide
respectively.
Sections and smears prepared from birds killed at the end of fifth day, were
submitted to the action of ribonuclease [Kunitz, 1950] and then stained according
to Brachet's method (loc. cit.) to differentiate between the metachromasy due to
mucoid sulphate and ribonucleic acid.
OBSERVATIONS
A. Distribution of mucoid sulphate
Sporozoites lying free in the lumen and freshly penetrated ones contained a
few violet† granules (metachromatic) round the nucleus (Plate XII, fig. 5). Some-
times a few more granules were seen lying in the form of a string between the nucleus
and the siderophilous body.
First generation schizonts in addition to the blue orthochromatic staining which
they took more intensely than the host tissue, also showed definite violet granules
embedded in purplish cytoplasm. Merozoites liberated after their rupture showed
metachromatic violet staining area at the acute (anterior) end, very rarely meta-
chromatic substance was noted at the opposite end also.
* At present Professor of Protozoology, School of Tropical Medicine, Calcutta.
† In addition to revealing the orthochromatic substances in the protoplasm, toluidine blue has the
property of staining the mucoid sulphate as purple, violet or pink (metachromasy).
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