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SOME EFFECTS OF FEEDING WHITE RATS WITH RATANJOT
(ONASMA ECHIODES) IN VAN ASPATI AND ITS
PATHOLOGICAL SIGNIFICANCE.
By D. A. MUNRO and J. M. LALL, Indian Veterinary Research Institute, Izatnagar
(Received for publication on 6 July 1948)
T HE powdered leaves and roots of the plant Onasma echiodes Linn. (ratanjot) are
in common use in ayurvedic medicine for the treatment of various affections
in the human subject and although much has been learned of the therapeutic use
of this drug, little is known of its activity in the body after absorption. It has
been suggested that the dye extracted from the roots of the plant would be a suitable
agent for colouring vanaspati to distinguish it from ghee when retailed for sale and,
in furtherance of this aim, it was considered desirable to test its toxicity on small
animals. In studies carried out at this Institute [Pande and others] results have
shown the coloured vanaspati to be non-toxic when fed to rabbits, as no symptoms
of toxicity developed. It was, however, considered advisable to reinvestigate this
problem in order to determine if the same factors had any influence on white rats,
and whether the addition of the dye to vanaspati rendered it toxic by its repeated
and continuous absorption in the tissues of the body over a prolonged period and
if so, whether it was sufficient to produce symptoms of clinical importance. White
rats were chosen for this study, as these animals are mostly used for dietetic experi-
ments on account of their adaptibility to a variety of diets; besides in feeding habits
they approximate most closely to man in subsisting on foods compounded from
different sources. In an attempt to use rabbits for our feeding experiments in the
early stages of this work it was found that when ratanjot was mixed with vanaspati
and added to the diet, the feed was totally refused, or eaten very sparingly. Feeding
with the aid of a graduated pipette was then resorted to, but here, there was too
much resistance on the part of the animal, and the danger of the food entering the
trachea and bronchii was always considered a risk, besides a good deal of wastage
occurred making it impossible to estimate accurately the total quantity taken. An
appreciation of these facts led us to abandon the use of rabbits, as bad results are
likely to follow when food unfit for a particular type of animal is forcibly given.
A distaste for the coloured vanaspati or as a matter of fact for vanaspati alone when
fed by drenching showed itself by the animal eating very sparingly and often starving
itself, thus affording an excellent example of the bad results which follow the abuse
of natural laws in connection with dietetics. On the other hand the feeding of rats
with coloured vanaspati and vanaspati alone is quite a simple affair, the rats accustom
themselves gradually to this diet and no interference with the normal process of
digestion is brought about.
11
D
12
SOME EFFECTS OF FEEDING WHITE RATS WITH RATANJOT [Vol. XIX, Part I
METHOD
Concentrated ratanjot in powdered form was obtained from the Director,
Indian Dairy Research Institute, Bangalore. A solution of the dye in vanaspati was
prepared by gently heating the mixture repeatedly till as much of it as possible was
dissolved. This was filtered through cotton wool and the concentrated dye used to
colour the vanaspati to produce colour readings of 5, 12, and 25 red units in a half
centimeter cell in the Lovibond tintometer. The experiment was carried out on
eight rats and two control rats, all within the 85 to 90 days age limit. The rats were
divided into four groups and all were kept on the same stock diet consisting of wheat
flour 65 per cent, maize 15 per cent, casein 10 per cent, wheat bran 3 per cent, salt
1 per cent, yeast 5 per cent and calcium carbonate 1 per cent, plus an allowance of
milk and green daily. In addition, raw goat liver was given twice weekly. The
progress of each animal was watched and each was weighed every ten days. Mea-
sured quantities of coloured vanaspati calculated at the rate of 1.25 c.c. per 100 gram
body weight was given orally once daily to each rat with the aid of a graduated
pipette as follows:
Group 1.—Two rats with a solution of the dye in vanaspati with a colour reading
of 5 red units and heated to 160-170°C.
Group 2.—Two rats with a solution of the dye in vanaspati with a colour reading
of 12 red units and heated to 160-170°C.
Group 3.—Two rats with a solution of the dye in vanaspati with a colour reading
of 25 red units and heated to 160-170°C.
Group 4.—Two rats with a solution of vanaspati alone heated to 160-170°C.
At four months from the commencement of the experiment the animals were
killed and post-mortems conducted.
CLINICAL HISTORY
In general, it may be said that the symptoms which develop are the outcome of a
rich generous diet which in no way impaired the condition of the animal. The
large intake of vanaspati in any form was followed by a depressed appetite, marked
decrease in activity and a greater laying on of adipose tissue. The animals grew
well and the weight increase was progressive till the end of the experiment. No
appreciable differences in weight and normal body size of these experimental animals
as compared with other rats of the same age groups in our rat colony could be detected.
It will be noticed that no significant symptoms were obtained, although slight toxicity
might easily exist in the dye without recognition, so that these cases are more of
interest to the pathologist than to the clinician.
POST-MORTEM FINDINGS
There was a great increase of fat in the subcutaneous connective tissue and in
the folds of the mesenteries of rats receiving the coloured vanaspati and vanaspati
alone. The kidneys of all the rats were covered over with fat, but no alterations in
naked eye appearance were seen. In some of the rats of Groups 1 to 4, the
lungs were seen to contain small nodules which varied in size and number, at times
March, 1949.]
D.
A.
MUNRO
AND
J.
M.
LALL
13
coalesing to form a small mass. On microscopic examination Gram negative cocci-
form bacilli were seen which confirmed the identity of the disease to that caused by
Bacterium pseudotuberculosis rodentium. The most striking feature at post-
mortem was that in the livers of rats receiving the coloured vanaspati, the surface
was studded with haemorrhagic spots giving it a spotted appearance. These lesions
were seen to extend to the parenchyma. Varying degrees of differentiation were
observed in the livers of those receiving more of the dye, which also showed a wider
range of changes on microscopic examination. Macroscopically, the livers on cross
section showed a deeper reddening of the hepatic parenchyma with dilated central
and peripheral veins. All other organs were normal.
HISTOLOGICAL FINDINGS
A histological examination of the tissues of all rats under the experiment was
carried out. The organs examined were the heart, lungs, liver, spleen, kidney,
stomach, intestines, and eye. The examination showed very few changes in the
tissues examined. Slight changes were found in the spleens of rats receiving larger
amounts of the dye which showed congestion and dilatation of the venous sinuses.
Patches of broncho-pneumonic lesions were frequently found in some of the lungs
indicating a secondary infection, but as these lesions had no relation whatsoever
with the experiment itself, they did not merit further consideration. The main
changes were in the liver of rats receiving the coloured vanaspati, details of which are
given below.
Rats in Group 1
A slight widening of the hepatic capillaries with no changes in the liver cells.
Rats in Group 2
Hepatic capillaries engorged. Liver cells are seen to disintegrate, fade and
gradually disappear, leaving pale areas devoid of liver cells.
Rats in Group 3
The hepatic capillaries are more deeply engorged. The liver substance is,
congested, although the inflammatory process is not evenly distributed. Here too
the liver cells are seen to disintegrate and fade, leaving in some parts of the hepatic
tissue pale areas completely devoid of liver cells.
DISCUSSION
It will be seen from the above that the most important changes are brought
about in the liver, following the presence of an irritant in the portal blood. It is
an interesting fact that the absorption of the dye after a period of feeding for four
months does not cause any special symptoms, although the morbid change affects the
liver of the animals in varying degrees. Further, it does appear entirely improbable
that the presence of these lesions in the bodies of the rats should be accidental,
especially since the gradations of the lesions in the three groups of rats vary in
severity with the amount of the dye fed.
14
SOME EFFECTS OF FEEDING WHITE RATS WITH RATANJOT [Vol. XIX, Part I,
SUMMARY
The feeding of ratanjot dissolved in vanaspati (with a colour reading of 5 red
units in a half centimeter cell under Lovibond tintometer and heated to a tempera-
ture of 160-170°C) for a period of four months did not reveal any toxic symptoms,
but on autopsy hæmorrhagic spots were present under the capsule of the liver
which might also be seen in the parenchyma. No changes in the liver cells were
observed.
In the case of rats of groups 2 and 3, which were fed continually for a period
of four months with a higher concentration of the drug having a tintometer reading
of 12 and 25 red units, the livers showed definite changes, with severe destruction of
liver cells. Ba d on the evidence obtained from feeding rats with ratanjot dissolved
in vanaspati with a colour reading of 5 red units, it would appear to be advisable to
use a much lower concentration of the dye to tint vegetable fats to avoid the harmful
effects on the liver, if it is desired to use this substance as a colouring agent. No
abnormal charges were observed in the livers of the control rats.
ACKNOWLEDGEMENT
We wish to express our thanks to Dr N. D. Kehar for his great interest and
assistance in this work.
THE RELATIONSHIP BETWEEN THE DISTRIBUTION OF
THE OX WARBLE-FLY (HYPODERMA LINEATUM DE
VILLERS) AND SOIL MOISTURE IN INDIA
By B. N. SONI, B.Sc.(PUNJAB), B.Sc. AGRIC. (EDIN.), Officer-in-Charge (Hides and
Skins Schemes), Indian Veterinary Research Institute, Mukteswar-Kumaun
(Received for publication on 5 July 1948)
T HE outstanding fact in the regional distribution of warble flies is their occurrence-
in dry, arid localities and their rarity in, or almost complete absence from,
moist areas. Thus, Bishop and his collaborators [1926] working in the States of
America, state that warble grubs are not found in cattle in low swampy areas; whilst,
according to Smith [1932], they are more numerous in Kansas after a dry spring
than after a wet one. Walton [1925], in his account of warble-fly infestation among
cattle in Great Britain observes: 'Tabulated records suggest a diminution in the
abundance of warble larvae following a cool, rainy spring in the preceding year.'
In India, Fletcher and Sen [1930] refer to the infestation being 'as a rule, restricted
to North-Western India, and absent from the moist areas, such as Madras, Assam,
Burma and Ceylon'. Numerous observations made by the present writer in the
course of his warble-fly surveys of different localities in India, point to the same
conclusion.
Thus, during the winter of 1938 the writer examined 178 heads of cattle of the
Dacca Government Cattle Farm in Bengal and none of these showed warble larvae on
the back, in spite of the fact that for some previous years warble-infested bulls
had been imported into Dacca from Hissar (Punjab). A similar observation was
recorded at Trivandrum in South India. Here the writer examined about 20 dairy
cows recently imported from Karachi and all showed fully developed H. lineatum
larvae in their backs, the infection in these cases having been apparently acquired
before importation into Trivandrum; for no tumours were found on local breeds of
cattle or on cows previously imported from Karachi. During 1937-38, the writer,
in collaboration with Mr S. R. Chadha, carried out a survey of the North-West
Frontier Province and the infestation was found to be 'very heavy in the hilly
districts.' In places where the land is marshy all along, e.g., in Bannu, the warble-
fly is very rare but in the dry sandy areas like Dera Ismail Khan it has been most
abundant [Chadha and Soni, 1939]. Likewise, in Rajputana, 5,355 (68.65 per cent)
heads of cattle were found infested out of a total of 7,800 examined there by the
writer in November 1941, whilst in certain tracts not one animal was found free
from the infestation. It is noteworthy that similar observations have also been
recorded in the case of the goat warble-fly (H. crossii), for Cross and Patel [1921],
*Paper read at the 24th meeting of the Indian Science Congress, held at Banaras in
January, 1941. Re-written in the light of the discussion which took palce at the
meeting.
15
E
March, 1949.]
B.
N.
SONI
17
for out of a batch of nine pupae thus treated six fully developed adults emerged, the
average pupal period in these cases varying from 14 to 18 days. A constant tem-
perature of 22°C. was maintained throughout these experiments. These results
are summarized in the following Table I.
TABLE I
Observations made on H. lineatum larvae and pupae kept at known moisture conditions
Percentage of soil moisture
20 . . . . . . . . . .
15 . . . . . . . . . .
10 . . . . . . . . . .
1 to 5 . . . . . . . . . .
Percentage
of
emergences
nil
nil
16.6
66.6
Pupal period
..
..
42 days
14 to 18 days
The laboratory observations described above are claimed to demonstrate con-
clusively the intimate relationship of soil moisture with the pupal development of
H. lineatum and indirectly with the regional distribution of the pest. It is of interest
that they are in accord with the results of some field experiments carried out by
Bruce [1938] in the United States of America, for these results 'showed that although
Hypoderma pupae would not develop in wet soil, the same soils, provided with ample
drainage, would permit complete development and adult emergence.'
SUMMARY
An excess of soil moisture has been found to have detrimental effect on the
development of the Hypoderma lineatum in its pupal stage, which normally extends
over a period of nearly six weeks. Experiments carried out in the laboratory at
Mukteswar have shown that the maximum number of emergences occur at a soil
moisture varying from 1 to 5 per cent. No fly emerged from pupae kept at a soil
moisture of 15 per cent and above. It is concluded that, it is the moisture content
of the soil that accounts for the concentration of the H. lineatum in certain localities
and its absence from others. This conclusion is borne out by field observations on
the regional distribution of the pest in India and in the United States of America.
ACKNOWLEDGEMENTS
It is my pleasant duty to express my grateful thanks to Dr S. Datta, Officer-in-
Charge, Veterinary Zoology Section (now Section of Parasitology) of the Institute
(at present Director of the Institute) for his keen interest during the course of this
investigation. I am also greatly indebted to Rai Bahadur S. K. Sen, Research
Officer (Entomology) (now retired), for useful suggestions.
18 THE RELATIONSHIP OF OX WARABLE-FLY AND SOIL MOISTURE [Vol. XIX, Part I,
REFERENCES
Bishopp et al. (1926). U. S. A. Dept. Agric. Bull. 1369
Bruce, W. G. (1938). J. econ. Ent., 31, 639-642
Chadha, S. R., and Soni, B. N. (1939). Indian J. Vet. Sci. 9, 101-103
Cross, H. E., and Patel, P. G. (1921). Dept. Agric. Punjab Vet. Bull. 3
Fletcher, T. Bainbrigg, and Sen, S. K. (1930). J. Cent. Bur. Anim. Husb. & Dairying Ind., 4,
127-138
Smith, R. C. (1932). J. Kans. ent. Soc, 5, 65-92
Walton, C. L. (1925). Welsh. J. Agric. 1, 195-199
THE NUTRITIVE VALUE OF THE INDIGENOUS GRASSES
OF ASSAM
The aquatic grasses, their chemical composition and nutritive value
By S. K. TALAPATRA, and M. N. DEV GOSWAMI, Animal Nutrition Research Scheme,
Assam, Gauhati
(Received for publication on 14th June 1948.)
T HE importance of the uncultivated grasses in the economic feeding of our
cattle needs no special emphasis. In the eastern provinces of the country
where conditions are favourable, they seem to grow in abundance during their
season time. Organised attempts have not been made to utilise these grasses as
cattle feeds, partly because little is known about their palatability, chemical com-
position and nutritive value. However, data on the composition of some of the
highland indigenous grasses are available [Sen, 1937] and [Kehar, 1947] but there
seems to be no reference in the literature either in regard to the chemical composition
or the nutritive value of the aquatic grasses which grow in abundance almost in
every part of Assam.
In view of the acute shortage of fodder in the country, it has been considered
desirable to explore the possibilities of utilising these stuffs as cattle feeds. We have
recently completed some detailed investigations as to whether the aquatic grasses
can be profitably utilised in the dietary of our cattle as green fodder, hay and silage.
The object of the present communication is to give an account of these investiga-
tions as follows:—
I. The chemical composition with progressive maturity as represented
by the monthly cut samples under different soil conditions.
II. The metabolism of the organic and inorganic nutrients as influenced by
feeding the grasses as green fodder, hay and silage.
The aquatic grasses which can be collected in large quantities are of two kinds
commonly known as dal (Hymenachne amplexicaulis Nees) and uridal (Oryza sativa
var. Fatua) in this part. The dal contains a spongy pith, while the uridal is free
from it. Even when fully mature, they are leafy, soft and succulent. It has been
estimated that the yield of these grasses from a medium size dalani (lowlying area)
of 500 acres is about three lakh maunds per year.
CHEMICAL COMPOSITION
To study the chemical composition on a monthly cut system, samples of grasses
were collected from three different places namely, Khanapara, Tongra and Panja-
bari in case of dal, while uridal was collected from Beltola. For the sake of conveni-
ence and better control in the method of sampling, etc., these areas which were quite
close to our laboratory were selected. The chemical analyses have been done
according to A. O. A. C., 1945 edition and Talapatra S. K., Ray S. C., Sen K. C.
[1942]. The results are shown in the following Tables.
19
F
20 THE NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XIX, Part I,
TABLE I
Protein content of the monthly cut samples on dry basis
Month
April . . . . . . .
May . . . . . . .
June
.
.
.
.
.
.
.
July . . . . . . .
August . . . . . . .
September . . . . . . .
October . . . . . . .
November
.
.
.
.
.
.
December . . . . . . .
Dal grass
Khanapara
27.1
15.6
14.8
14.1
14.0
..
13.8
13.1
9.3
Tongra
28.1
17.8
10.8
11.5
11.0
..
10.8
10.6
9.4
Panjabari
29.4
14.1
13.1
11.9
10.6
13.4
13.8
10.4
10.8
Uridal grass
Beltola
..
16.2
13.5
14.7
12.0
12.2
11.8
10.5
10.2
The data in the above Table will show that the aquatic grasses are rich in protein.
Even in December when most of the grasses turn woody and fibrous, the aquatic
grasses retain as much protein as can be found in some of the cultivated fodder
plants like maize, jowar, Napier and Guinea at their early stages.
TABLE II
Calcium content of the monthly cut grasses on dry basis
Month
April . . . . . . .
May . . . . . . .
June . . . . . . .
July
.
.
.
.
.
.
August . . . . . . .
September
.
.
.
.
.
October . . . . . . .
November . . . . . . .
December . . . . . . .
Dal grass
Khanapara
0.332
..
0.300
0.303
0.270
..
0.210
0.141
0.130
Tongra
..
0.29
0.26
0.29
0.23
..
0.24
0.27
0.20
Panjabari
0.240
0.240
0.280
0.271
0.270
0.260
0.220
0.200
0.140
Uridal grass
Beltola
..
0.401
0.380
0.300
0.340
0.330
0.290
0.270
0.250
March, 1949.] S. K. TALAPATRA AND M. N. DEV GOSWAMI 21
The results in Table II show that there is little variation in the content of calcium
between the samples collected from the three different places. Compared to dal,
the uridal is definitely richer in calcium. With the progressive maturity, the content
of this mineral decreases. Both the grasses may be regarded as poor in calcium.
TABLE III
Phosphorus content of the monthly cut grasses on dry basis
Month
April . . . . . . .
May . . . . . . .
June . . . . . . .
July . . . . . . .
August . . . . . . .
September . . . . . . .
October . . . . . . .
November . . . . . . .
December . . . . . . .
Dal grass
Khanapara
0.294
..
0.300
0.312
0.281
..
0.200
0.210
0.160
Tongra
..
0.541
0.382
0.391
0.342
..
0.351
0.260
0.210
Panjabari
0.441
0.490
0.341
0.331
0.292
0.291
0.211
0.220
0.200
Uridal grass
Beltola
..
0.422
0.382
0.300
0.342
0.335
0.290
0.251
0.210
The data shown in the above Table are self-explanatory. The grasses seem to
be rich in phosphorus, specially at their early stages. With the progressive maturity,
the content of phosphorus also decreases and is lowered by almost half at the flowering
stage. Samples were also collected from the various representative tracts of the
province and analysed. The results are shown below in Table IV.
F 2
22 THE NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XIX, Part I,
TABLE IV
Chemical composition of the grasses from all over the province on dry basis
Localities
Upper Assam
Central Assam
Lower Assam
Upper Assam
Central Assam
Lower Assam
1 . .
2 . .
3
.
.
1 . .
2
.
.
3 . .
1 . .
2
.
.
3 . .
1 . .
2 . .
3 . .
Crude protein
Calcium
Dal grass
15.2
14.4
8.3
12.2
11.1
11.2
21.9
11.0
14.9
12.8
11.5
(pit-soil)
12.9
0.20
0.30
0.19
0.30
0.37
0.30
0.15
0.11
0.11
0.23
0.32
0.11
Uridal grass
12.0
11.6
12.4
0.27
0.25
0.19
Phosphorus
0.28
0.25
0.17
0.35
0.33
0.351
0.16
0.29
0.31
0.21
0.22
0.21
0.23
0.34
0.30
A perusal of the data in Tables I to IV will show that while the content of protein
is fairly constant irrespective of the places from which samples were analysed, the
content of calcium varied from 0.11 in the case of Lower Assam to 0.32 in central
Assam. It may, therefore, be resonably concluded that—
(a) the aquatic grasses are fairly rich in protein;
(b) they seem to be poor in calcium;
(c) they are not deficient in phosphorus.
NUTRITIVE VALUE OF THE AQUATIC GRASSES
Although the aquatic grasses are quite rich in some of the indispensible nutrients
like the protein and phosphorus, contrary to expectation, they are not much relished
by cattle, specially at the early stages when fed as green fodders. It appears that
due to some fishy smell, our cattle refuse these stuffs. Whatever may be the cause,
as a result of this practical difficulty, we were about to abandon our plan of investiga-
tions, but soon it was realised that at least one of these namely, the dal grass could
be fed to appetite at the flowering stage as a green fodder. But soon after the trial
it was found that if the aquatic grasses were fed green, cattle suffered from worm
trouble and that the feeding of these grasses to young animals led to serious conse-
quences.