NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
N. S. DOCTOR, B. N. BANERJEE AND Z. R. KOTHAVALLA 77
but there was no increase in fat content. This effect, though marked and
characteristic, may not be very pronounced as a number of other factors such
as breed, season and lactation period also influence the composition of the
milk fat.
The effect of feeding oil cakes on the nutritive value of ghee, as measured
from its carotene and vitamin A content, is also marked. The feeding of
linseed cake decreases the vitamin A and carotene content considerably but
both increase with foods rich in carbohydrates and with green grass.
In studying the effect on ghee, the results obtained show that cows
of different breeds exhibit differences in their abilities to transfer carotene
from the food into their butterfat. The butterfat of the foreign breeds con-
tains more carotene than that of Indian breeds. Even the butterfats from
individuals of the same breed of cow fed on the same rations show marked
differences in carotene and vitamin A content. Buffalo butterfat differs in
this respect from cow butterfat. The physiological mechanism of the buffalo
appears to cause very little secretion of carotene into its milk fat.
From these results, it is evident therefore that the excessive feeding of
oil seed and cakes must affect the composition, texture and flavour of butterfat.
The results indicate that the composition of the milk fat of animals fed on a
particular oil seed or cake tends to become similar to that of the oil. How-
ever, the results of a short period experiment, while indicating the way the
quality of butter and ghee is affected, cannot be taken strictly to represent
any herd, seasonal or regional variation without limitations.
Standards for grading of ghee.—The problem of fixing grading standard
for particular types of ghee is difficult in spite of much available data and
literature on the subject. The main reason is that the range of variation is
very wide and depends on so many factors, such as climate, breed and care of
the animals, feed, idiosyncracy of individuals, period of lactation, etc. Further,
the problem is complicated in a country like India, where large quantities of
ghee are being prepared by small stock owners, owning only one or two cows
or buffaloes, generally half-starved animals kept under unhygienic conditions.
Under these conditions no standard of grading, however liberal, will prove
useful as an all-India grading standard under all circumstances. However,
to facilitate the task of the public analyst the data obtained in this experiment
will be of value in correlating variations due to breed, feed and method of
preparation of ghee.
The limits of variation of various constants under the condition of this
experiment are as follows :—
B. R. reading at 40°C...
Reichert value
....
Polenske value
....
Kirschner value
....
Saponification value . ...
Iodine value
.....
Percentage of free fatty acids calculated
as oleic acid
....
Cow
40—45.2
23—30
1.9—3.0
17.0—24.2
219—230
29.0—42.3
0.21—0.91
Buffalo
42.5—43.6
30.2—31.7
1.5—2.0
26.6—26.8
229—239
30.8—34.9
....
L
78 Factors affecting Physical & Chemical Constants of Ghee
The most debatable constant in the grading standard is the Reichert
value of ghee. The Reichert value indicates the extent to which the steam-
volatile, water-soluble fatty acids (low molecular weight acids mostly butyric
to caproic) are present in fat. Butterfat or ghee contains much higher
amounts of these acids than any other fat from which it might be desirable
to distinguish it. Hence the fixing of this value requires very careful and
accurate data. Further, in view of the results obtained with regard to feeding
oil seeds and cakes, the importance of the iodine value should not be over-
looked. Smith and Dastur [1938] have recorded a marked fall and rise in the
Reichert and iodine values respectively in the milk fat of fasting cows. In
the first two days of fasting the Reichert value fell from 26 to 14.5 and the
iodine value rose from 36 to 47 .1. Various workers have observed a correlation
beween the iodine value and refractive index of ghee. The figures tabulated
by Elsdon [1926], those obtained by Kothavalla and Doctor [1939] and in
this experiment are conspicuous in this respect. Hence the iodine value
should also be incorporated in the specified standards for testing purity and
the grading of ghee.
The results obtained in the present investigation and from the data
obtained by other workers [Bhattacharya and Hilditch, 1931 ; Elsdon, Taylor
and Smith 1931 ; Bolton and Revis, 1911 ; Sunawalla and Kothavalla, 1935;
Trieman, 1913 ; Ghosh, 1920 ; Kothavalla and Doctor, 1939] for the analytical
values of cow and buffalo ghee show the range of variation for the various
constants as follows :—
Reichert value . .
Polenske value . .
Kirschner value . .
Iodine value . .
B. R. reading at 40°C .
Cow ghee
Not less than 23
1.5—3.0
Not less than 19
26—38
40—46
Buffalo ghee
Not less than 30
1.5—3.0
Not less than 25
30—38
40—46
It does not, therefore, follow that any sample of ghee which does not
fall within this range is adulterated or artificially blended, and that which
is within the range is a genuine one. It is of importance, therefore, when
coming to a decision with regard to butter or ghee which appears to be
adulterated with small quantities of vegetable or animal fats, to obtain,
wherever possible all the information available regarding the history of the
sample, as it is not reasonable to express the opinion that the abnormal sample
should always be considered as adulterated. The cumulative evidence of
all constants and the history of the sample should, therefore, be taken into
consideration in judging the quality or purity of ghee and butter.
SUMMARY
The effect of methods of preparation, breed and feeding of oil cakes to
dairy cattle on the physical and chemical constants of ghee has been described.
Methods of preparation of ghee have no appreciable effect on the Reichert
value and other constants of ghee.
N. S. DOCTOR, B. N. BANERJEE AND Z. R. KOTHAVALLA 79
Different breeds of cattle and even individuals of the same breed differ
in their ability to transfer carotene in the milk from the food into their butter-
fat.
The composition of the milk fat of animals fed on oil cakes tends to become
similar to that of the oil.
Groundnut, linseed, gingili cake and cotton seed have been shown to
increase the iodine value and decrease the Reichert value while coconut
cake decreases the amount of unsaturated acids but has no effect on the
Reichert value.
The quality of butter produced by feeding linseed and coconut cake has
been shown to be improved while that of ghee to be deteriorated because
of the oily flavour imparted by the cakes.
The feeding of oil cakes has a marked effect on the body and texture of
butter and ghee. Gingili cake produces a soggy soft butter and ghee.
The carotene and vitamin A contents of butterfat decrease slightly on
feeding oil cakes, and increase with food rich in carbohydrates and on green
grass. Foods rich in carotenes appear to give a higher carotene content in the
milk fat.
The effect of feeding oil cakes, breed, and method of preparation on the
composition of ghee has been discussed, and the range of variation of its
physical and chemical constants has been defined.
REFERENCES
Banerjee, B. N., and Doctor, N. S. (1938). Agric. & Livestock in Ind. 8 , 158
Bhattacharya, R., and Hilditch, T. P. (1931). Analyst 56, 161
Bolton, E. R.. and Revis, C. (1911). Analyst 36, 372
British Standard Methods for the Chemical Analysis of Butter, 1938, Brit. Standards
Inst. 769
Brouwer, E. (1931). Vereen Exploit. Proefzuvelboerd. Hoorn. Ann. Rept. 113
Channon, H. J., Drummond, J. C., and Golding, J. (1924). Analyst 49, 311
Cranfield, H. T. (1911). Analyst 36, 445
–––––––––––
(1916). Analyst 41, 336
Dean, H., and Hilditch, T. (1933). Biochem. J., 27, 889
Eckles, C. M., and Palmer, L. S. (1917). Analyst 42, 389
Elsdon, G. D. (1926). 'Edible Oils and Fats'
Elsdon, G. D., Taylor, R. J., and Smith, P. (1931). Analyst, 56, 515
Ferguson, W. S., and Bishop, G. (1936). Analyst 61, 515
Ghosh, T. K. (1920). Analyst 45, 444
Hilditch, T. P., and Sleightholme, J. (1930). Biochem. J. 24,1098
––––––––––––––––––––––(1931).
Biochem. J. 25, 507
–––––––––––and Thompson, H. M. (1936). Biochem. J. 30, 677
Hunziker, O. F., Mills, H. C, and Spitzer, G. (1912). Ind. Agric. Expt. Sta. Bull. 159,
311
Holland, E. B., and Buckley, et al, (1918). J. Agric. Res. 12, 719
––––––––––––––––––––––(1923).
J. Agric. Res. 24, 365
Kothavalla, Z. R., and Doctor, N. S. (1939). Ind. J. Vet. Sci. & Anim. Husb. 9, 151
80 Factors affecting Physical & Chemical Constants of Ghee
Lyons, J. (1936). Economic Proc. Royal Dublin Soc. 3, 2, 17
Mohr, W. (1925). Milchw. Forsch. 2, 24
Neasham, E. W., and Gelpi, A. J. (1934). Nat. Butter & Cheese J. 12
Platon, B., et al, (1935). Rept. Centr. Agric. Res. Inst., Stockholm 451
Report of the Analytical Methods Committee, (1936). Analyst 61, 404
Siegfeld.M. (1907). Analyst 32, 256
Smith, J. A. N., and Dastur, N. N. (1938). Biochem. J. 32, 1868
Sunawalla, S. D., and Kothavalla, Z. R. (1935). Agric. & Livestock in Ind. 5, 480
Trieman, S. H. (1913). Analyst 38, 247
Wright, N. C. (1937). Rept, on the development of Cattle and Dairy Industry in India,
159
SOME OBSERVATIONS ON THE VIABILITY OF DOYLE'S
DISEASE VIRUS OF FOWLS UNDER DIFFERENT
CONDITIONS*
BY
S. GANAPATHY IYER, G.M.V.C.
Imperial Veterinary Research Institute, Mukteswar
(Received for publication on 10 February 1939)
INTRODUCTION
D OYLE'S disease is an acute, highly contagious and infectious disease of
poultry, known to the veterinary profession since 1926. It is
caused by a filterable virus. This disease was observed in England,
Dutch East Indies, Philippine Islands, India, Ceylon, Korea, Japan
and Australia, but little information as regards the source from whence
the disease was introduced has been forthcoming.
The nomenclature of this disease has caused some confusion, various
names having been given during recent years for the same disease. For
convenience, the disease has been termed Doyle's disease in this paper.
A knowledge of the survival of the virus outside the animal body is a
necessity in the control of epizootics. This paper records the results of experi-
ments conducted with reference to the viability of the virus under labora-
tory conditions as well as a review of the available literature on the subject.
The laboratory where this work was done is situated in the Kumaun hills of
the United Provinces, at an altitude of 7,500 ft. and the climate is temperate.
It is believed that the virus, under climatic conditions such as obtain
in the plains of India, will seldom remain alive for periods longer than those
recorded in this paper. Only scattered information on the subject is avail-
able from literature. Doyle [1927] in his first paper on the discovery of a
new virus causing this disease among fowls, ' Newcastle Disease ', recorded
that the virus in dried tissues was mostly viable by the 100th day. Virus
in saline was active up to 86 days but inactive at the 100th day.
Spleen virus as kept in a petri-dish was viable after 80 days but dead
after 150 days. While studying the effects of heat, in the only experiment
carried out, the virus was viable after 48 hours at 37° C., but was
inert after 72 hours. The same author [1935] considers that the
virus will remain alive for at least six months in the muscles and bone marrow
of carcases under ' trade chilling conditions '.
Picard [1928] found that indirect contact by placing eight healthy fowls
in a small coop in which ten fowls had previously died successively and no
cleaning was done, resulted in the death of three fowls and the above experi-
ment was repeated on a smaller scale. The fowls were left in a cage soiled
with faeces and discharges of a few sick fowls that died 14 days pre-
viously. Nothing happened to the fowls for six weeks, thus indicating that
*Paper read at the Indian Science Congress, 1939.
(81 )
82
Viability of Doyle's Disease Virus of Fowls
the virus had died out. Saliva from sick fowls, diluted 1: 10, was stored
in a tube plugged by cotton wool and after exposure to full day-light, it was
shown that the virus was dead after 24 hours. Brain substance
dried at 27° C. over quick lime was active for 35 days.
Farinas [1930] stated that direct sunlight killed the virus after one-hour
exposure. Spleen and liver in a petri-dish covered with earth and stored at
room temperature for 12 days, when fed to chickens, produced no disease.
Virulent crop contents absorbed by cotton wool and allowed to dry at room
temperature became a virulent in five days. Virus in dilute crop contents by
storing at 37.5°C. was also rendered aviable after three days. Virus in
garden soil under experimental conditions after exposure for four days to
diffused sunlight did not produce infection by feeding to chickens. The
virus was suspended in stagnant water and after four days the virus was
found to be dead. This writer also found that active virus could be recovered
from a bird seven days after death.
Kylasamier [1931] reports that cotton wool swabs charged with throat
material by officers in the field and sent by mail to Madras, the journey period
being two to three days, were invariably active on animal inoculation.
MATERIALS AND METHODS
Liver and spleen tissues from fowls that died of Doyle's disease after
experimental inoculation with passaged virus was used for the different tests
except where otherwise stated. The virus had undergone over 70
fortnightly passages in fowls with no material alteration in virulence. About
1 gm of liver and spleen tissues was put into 2 c. c. test tubes plugged by
cotton wool. When due for animal inoculation, after the prescribed period,
the whole tissue from one tube was pulped and emulsified with 10 c. c.
saline Susceptible fowls were inoculated subcutaneously with 1 c. c. of
emulsion which roughly contained 0.1 gm of the tissue pulp. The fowls
were kept in separate iron cages during the period of observation. The
cages were kept in a shed away from the one used for routine Doyle's disease
work, with separate attendants and every possible care was taken to prevent
accidental infection. The aging of the virus was effected by storing the
tubes containing the tissue virus inside a glass cupboard and in the incubator
at 37°C. as the experiment required. The temperature of the room was
recorded twice a day, once at 9-30A.M. and again at 4-30 P.M. The maxi-
mum average daily temperature every month during the period when the
experiment was in progress, has been noted.
TABLE I
Average daily maximum temperature for each month during the course of the
experiment
July 1936
August 1936
September 1936
....
October 1936
....
November 1936
....
21° C.
21° C.
20° C.
17° C.
15° C.
S.
GANAPATHY
IYER
83
Viability of the virus after aging at room temperature.—The results of bird
inoculation tests (Table II) were summarized after the susceptibility of non-
reactors and those that survived after the experimental disease had been
tested. While drawing conclusions, it was borne in mind that aging might
attenuate virus to such an extent as to set up only a mild disease and thereby
confer protection to the fowls. Invariably, the fowls that escaped infection
from the original inoculation died when subsequently tested with fresh virus.
Thus it would appear that the change from viability to aviability is rather
abrupt. Owing to anomalous results, not unusual in experiments in virus
diseases, the viability tests had to be repeated in the same fashion but with
virus from different donors on different dates.
Periodical bird inoculations were made after aging from 3 days up
to 28 days. After the 17th day there was a notable drop in the
number of birds which became infected. Four fowls were inoculated with
virus after aging at room temperature for 21 days on different dates, but only
in one fowl was the disease reproduced.
TABLE II
Results of bird inoculation with virus after aging at room temperature
Virus
stored
for
(Days)
3
5
7
14
14
14
14
15
Date of
inoculation
25-10-1936
27-10-1936
13-7-1936 .
20-7-1936 .
15-9-1936 .
5-11-1936 .
16-9-1936 .
Fowl
No.
257
256
251
89
88
55
104
224
294
295
226
Result of inoculation
Original
Died or survived
Died
....
Do
Reacted and survived .
Died
....
Reacted and survived .
Died
....
Survived . . .
Died
....
Do
Reacted doubtfully and
survived
Survived . . .
Time
taken
to die
(Days)
16
18
...
6
...
10
...
12
12
...
...
Subsequent immunity
test with fresh
passaged virus
...
...
Survived . . .
...
Do. . .
...
Died.
...
...
Reacted and survived
Died
Remarks
Immune
Immune
84
Viability of Doyle's Disease Virus of Fowls
Results of bird inoculation with virus after aging at room temperature—contd.
Virus
stored
for
(Days)
15
16
17
18
18
19
20
21
28
Date of
inoculation
6-11-1936 .
17-9-1936 .
7-11-1936 .
18-9-1936 .
8-11-1936 .
19-9-1936 .
9-11-1936 .
20-9-1936 .
10-11-1936
21-9-1936 .
11-11-1936
27-7-1936 .
22-9-1936
12-11-1936
3-8-1936
Fowl
No.
297
296
227
299
298
228
300
301
229
302
303
230
310
161
311
120
119
162
313
140
139
Result of inoculation
Original
Died or survived
Survived . ..
Reacted and survived .
Do.
Survived . ..
Died
....
Survived
Died
....
Do
Survived
Do.
...
Died
....
Survived
Do.
...
Reacted and survived .
Survived . ..
Do.
...
Do.
...
Died
....
Survived . .
Do.
...
Do.
...
Time
taken
to die
(Days)
...
...
...
...
7
...
4
14
...
...
7
...
...
...
...
...
..
13
...
...
...
Subsequent immunity
test with fresh
passaged virus
Died.
Do.
...
Do.
Do.
...
Died
...
...
Survived .
Died
...
Died
Do.
None . . .
Died
Do.
Reacted and survived
...
Died
Do.
Do.
Remarks.
Susceptible even
after the ori-
ginal reaction
Immune
ori-
ginally
Paralytic symp-
tom
Viability of the virus after storage at 37°C.—Cotton plugged tubes contain-
ing the tissue virus were stored in the incubator at 37°C. The test inocula-
tions were made after three days and, at intervals of 2-4 days, subsequent
tests were continued until evidence was obtained that the virus had died
out.
S.
GANAPATHY
IYER
85
As will be seen from Table III, the virus was active after 24
days but apparently died out after 31 days when stored at 37°C.
In this connexion mention must be made of the appearance of the tissue
at the time of removal from the incubator. The tissue containing the virus
had lost much of its moisture content from the third day onwards. It seems
that the virus in desiccated tissues will remain viable for longer periods than
in saliva and crop contents where the virus was found to die out after three
days at 37.5°C. Farinas (loc. cit.) experiments with saline emulsions of
buccal washings from fowls dead of experimental Doyle's disease, stored at
37°C., showed that the virus remained viable after three days but was dead
after four days (not shown in Table III).
TABLE III
Effect of heat (37° C.) on the virus
Virus
stored
for
(Days)
3
5
7
7
9
9
11
11
13
13
17
21
24
31
Date of
inoculation
4-9-1936 .
6-9-1936 .
8-9-1936 .
29-10-1936
10-9-1936 .
31-10-1936
12-9-1936 .
2-11-1936 .
14-9-1936 .
4-11-1936 .
28-12-1936
27-12-1936
30-12-1936
6-1-1937
Test
fowl
No.
182
181
183
278
279
185
280
281
204
283
284
221
288
287
480
488
489
492
Result of inoculation
Original
Died or survived
Died .
Do. .
Do. .
Survived
Died .
Survived ....
Do.
Died .
Do. .
Survived
Died .
Survived
Died .
Do. .
Do. .
Do. .
Do. .
Reacted and survived
Time
taken
to die
(Days)
13
10
7
...
1½
...
...
12
9
...
11
...
12
8
6
23
18
...
Subsequent immunity
test with fresh
virus
...
...
...
Died
...
Reacted and survived.
Died
...
...
Died
...
Died
...
...
...
...
...
Survived
Remarks
Other causes
Viability of the virus exposed to sunlight.—The effect of direct sunlight on
the virus was studied at about 11 A.M. in the month of October. Ten c. c,
of a 1 in 10 organ emulsion (liver and spleen) in saline, free from gross particles.
M
86.
Viability of Doyle's Disease Virus of Fowls
was poured into a sterile petri-dish (4 in. diameter) and exposed to direct sun-
light. Healthy fowls were injected with this virus soon after the scheduled
time and the syringe barrel was covered with actinic paper during injection.
The work could not be extended on account of lack of sunlight. The virus
contained in organ emulsion seems to be unaffected in virulence after one
hour exposure to sunlight (Table IV). Longer periods seem necessary to
kill the virus, unlike the finding of Farinas (loc. cit.). This may perhaps be
due to geographical differences.
TABLE IV
Viability of the virus exposed to sunlight
Virus
exposed
to
sunlight
for
minutes
16
20
30
40
50
60
Date of
inoculation
24-10-1936
27-10-1936
Do. .
Do. .
Do. .
Do. .
Test
fowl
No.
254
255
272
276
269
268
275
277
273
274
154
270
Result of Inoculation
Original
Died or survived
Died
....
Reacted, paralysis sur-
vived
Reacted and survived..
Died .
Do.
.
Do.
.
Do.
Do.
.
Do.
.
Do.
.
Do.
.
Very late reaction sur-
vived
Time
taken
to die
(Days)
8
...
...
10
20
13
13
17
8
12
18
...
Subsequent immunity
test with fresh
virus
...
Survived .
Do
...
...
...
...
...
Died
Remarks
Immune
Do.
SUMMARY
The paper records the results of experiments conducted with reference
to the viability of the virus under laboratory condition as well as a review of
the past literature on the subject.
The virus used for the different tests was mostly contained in organs
such as liver and spleen. Ioyle's disease virus after aging at room tempera-
ture (about 17°C.) remained viable for 21 days.
At 37° C. the virus d d not die after 24 days but was inert after 31
days.
S.
GANAPATHY
IYER
87
Saline emulsion of buccal swabs from fowls dead of experimental Doyle's
disease gave variable results. It remained viable at 37°C. for three days
but was found to be dead after four days.
Exposure to direct sunlight for one hour had no lethal effect on the
virus.
REFERENCES
Doyle, T. M. (1927). J. Comp. Path. Ther. 40, 144-69
–––––––––––(1935).
ibid
48, 1-20
Farinas, E. C. (1930). Phili. J. Agric. 1, 311-364
Kylasamier, K. (1931). Ind. Vet. J. 7, 342
Picard, W. K. (1928). Dept. Agric. Ind. Commer. Dutch East Indies Vet. Bull. 65.
OBSERVATIONS ON THE PHYSIOLOGICAL
CONSTITUENTS OF EQUINE BLOOD*
BY
N. D. KEHAR, M.Sc., Sc.D.
Acting Officer-in-Charge, Animal Nutrition Section, Imperial Veterinary
Research Institute, Izatnagar
MOHAN SINGH, B.A., L.V.P.
Research Scholar, Government of the Punjab
AND
GOVIND RAO, B.A., G.B.V.C.
Serology Section, Imperial Veterinary Research Institute, Mukteswar
(Received for publication on 25 August 1939)
T HE lack of physiological data appears to be keenly felt in formulating a
scientific basis in modern therapy. It is believed that a sound knowledge
of the amount of the normal constituents of blood will be of incalculable value
in the diagnosis and treatment of several hitherto baffling diseased conditions
as a standard of comparison with samples from pathological cases. The
importance, therefore, of ascertaining the normal physiological standards of
the blood of domestic animals cannot be overemphasized as a fundamental
to our knowledge of pathology.
A considerable amount of work has been done in other countries on the
blood chemistry of domestic animals including horses, but a critical examina-
tion of the available literature reveals that no attempts have been made so
far in establishing these standards in India. Since heredity, diet, climate and
topography, besides other factors, are likely to introduce variations in the
blood constituents of animals in different parts of the globe as has been
observed in human beings [Sokhey, 1936], it was considered advisable to
attempt an investigation into the amount of sugar, creatine, creatinine,
calcium, inorganic phosphorus, chlorides, cholesterol and different protein
fractions of the blood of normal horses.
Fourteen apparently healthy and good-sized horses were selected for this
investigation. All of these animals were in a satisfactory condition of health.
Blood samples were collected by jugular puncture, unless otherwise stated,
in the morning after about 12 hours' fasting. All observations were made in
duplicate and the average values of two observations are given in Tables I
and II. Table III shows the maximum, minimum and the mean of the
average values.
* This work was carried out at the Physiological Laboratory (Serology Section),
Imperial Veterinary Research Institute, Mukteswar.
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