NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
STUDIES ON THE EFFECT OF VARIOUS FACTORS ON
THE VITAMIN C CONTENT OF THE MILK OF
COWS AND BUFFALOES
BY
ZAL R. KOTHAVALLA
AND
HARNAND SINGH GILL
Imperial Dairy Department, Bangalore
(Received for publication on 28 March 1942)
I N the human diet vitamin C is considered to be one of the vital factors in
the maintenance of health. Fresh milk is one of the sources for our supply
of vitamins. Since milk and milk products play a very important part in the
dietary of the majority of the Indian population, vitamin C is of special signi-
ficance. It is very sensitive to the action of light and heat, and in this country
where the milk is transported in open vessels for distribution, etc. and subject-
ed to high temperatures of boiling, before consumption, its study is made all
the more important.
Whitnah and Riddell [1937] found that the average values of vitamin C
content in milk for the different breeds of cows were, Ayrshire 24.1, Guernseys
26.0, Holsteins 23.5 and Jerseys 29.2 mg. per litre. Kon and Watson
[1936] observed that the vitamin C content of colostrum was slightly higher
than that of milk. The work of King and Waugh [1934] showed that the des-
truction of vitamin C in milk was much more in the ' holding method' than in
the ' flash method ' of pasteurization. In the same work the authors showed
that prolonged heating of milk, exposed to air, tended to destroy vitamin C
more severely than heating it to the boiling point for a short time. Kon and
Watson [1936] found that half an hour's exposure to sunlight was sufficient to
change all the ascorbic acid in a pint bottle of milk into the reversibly oxidized
form. Schwartze, Murphy and Hann [1930] showed that the rate of des-
truction of the vitamin C varied with the kind of metal with which the milk
came into contact during heating. It decreased by 20 to 40 per cent with
aluminium ; slightly greater with tinned copper and up to 80 to 90 per cent
with naked copper.
In India no work has so far been carried out to study the effect of various
factors on the vitamin C content of milk. The climatic and other conditions
of this country are so different from those of temperate countries that the
results obtained abroad could not be expected to hold good here. It was
therefore, thought that work done in this country on the lines followed by
workers in other countries might prove useful. Although this investigation
cannot be claimed to be an exhaustive one, it is hoped that the results obtained,
will prove useful for further investigation when so little data are available on
the subject in this country.
35
ZAL R. KOTHAVALLA AND HARNAND SINGH GILL 37
available. A representative sample was made up by taking 15 ml. of milk
from every sample. The vitamin C content of this sample was determined
on the lines indicated above. Daily variations in the values for each breed
of animals for 14 days, both morning and evening, have been determined and
the average figures for this period are presented in Table I.
TABLE I
The average vitamin C content of milk of cows and Murrah buffaloes as
determined before and after the hydrogen sulphide treatment
Class of animals
Sindhi cows
....
Gir cows . . . .
Crossbred cows....
Ayrshire cows....
Murrah buffaloes. . .
Vitamin C in mg. per litre of milk
Morning
Before
treatment
with H 2 S
26.4
26.3
23.0
22.7
27.0
After
treatment
with H 2 S
29.0
28.7
25.1
24.9
29.6
Evening
Before
treatment
with H 2 S
27.2
26.8
23.5
22.9
27.4
After
treatment
with H 2 S
29.3
28.9
25.6
25.0
29.8
It will be seen from Table I that of the four breeds of cows
studied, (a) the Sindhi cows produced milk of the highest and the Ayrshire
cows produced milk of the lowest vitamin C content; (b) the vitamin C con-
tent of the milk of Murrah buffaloes was also very high and higher than most
of the breed of cows ; and (c) the ascorbic acid content of the evening milk
was slightly higher than that of the morning milk.
(ii) Effect of breed on the vitamin C content of colostrum. The vitamin C
content of the colostrum of different breeds of cows and Murrah buffaloes
was also determined both morning and evening from the first to the fifth day
of calving. For the estimation of reduced and total ascorbic acid in the
colostrum, the same procedure as was adopted in the case of milk was followed.
TABLE II
The vitamin C content of the colostrum of cows and Murrah buffaloes
No. of
days after
calving
1
2
3
4
5
Class of animals
Sindhi cow
Vitamin C in mg.per litre of milk
Morning
Before
treatment
with H 2 S
27.3
27.6
27.6
27.0
26.4
After
treatment
with H 2 S
29.7
30.0
29.4
29.4
29.4
Evening
Before
treatment
with H 2 S
27.9
28.5
28.2
27.6
27.3
After
treatment
with H 2 S
30.3
30.6
30.0
30.0
29.4
38
Vitamin C Content of Milk of Cows and Buffaloes
TABLE II—contd
No. of
clays after
calving
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
Class of animals
Gir cow . .
Crossbred . .
Ayrshire . .
Murrah buffalo
Vitamin C in mg. per litre of milk
Morning
Before
treatment
with H 2 S
27.3
27.0
27.0
26.4
26.4
24.0
24.0
23.7
23.4
22.8
23.4
23.4
23.1
22.8
22.8
27.6
27.6
27.5
27.3
27 .3
After
treatment
with H 2 S
30.0
29.7
29.4
29.4
28.8
26.3
26.0
24.6
24.6
24.6
25.4
25.5
25.2
25 .2
24.9
29.9
29.9
29.7
29.7
29.7
Evening
Before
treatment
with H 2 S
27.6
28.2
27.6
27.0
27.0
25.5
25.2
25.2
24.6
24.6
23.7
23.4
23.4
23.4
23.1
27.7
27.7
27.6
27.6
27.3
After
treatment
with H 2 S
30.0
30.3
30.0
29.7
29.1
27.9
26.9
26.6
26.5
26.1
25.5
25.7
25.5
25.5
25.2
30.6
30.6
30.3
30.0
30.0
Table II indicates that: the ascorbic acid content of colostrum was
higher than that of milk.
(iii) Effect of pasteurization on the vitamin C content of milk. The
vitamin C content of milk was determined before and after it was pasteurized
by the ' holding ' and 'flash ' methods. In the ' holding method ' the milk
was heated to 145°F. for 30 minutes and after cooling it its ascorbic acid
content was estimated by titrating against 2 : 6-dichlorophenol indophenol
reagent. Four trials were made with this method of pasteurization. In the
' flash method ' the milk was heated to 165°F. and held at that temperature
for 30 seconds and then cooled and titrated for the vitamin C content. A
similar number of trials was made in this case also. In this as well as the
following experiments, no attempt was made to determine combined ascorbic
acid as it was time consuming and as the object was only to obtain compara-
tive data for the values of milk under various treatments. The figures given
in Table III represent only the reduced ascorbic acid contents.
ZAL R. KOTHAVALLA AND HARNAND SINGH GILL 39
TABLE III
The vitamin C content of milk before and after pasteurization
No. of
experi-
ments
1
2
3
4
Method of pasteurization
Holding . ..
Do.
...
Do.
...
Do.
...
Vitamin C in mg. per litre of milk
Vitamin C
before pasteu-
rization
25.6
25.2
25.8
24.6
Vitamin C
after pasteu-
rization
20.0
19.0
19.2
18.6
*Percentage
loss in pasteu-
rization
22.0
24.6
25.6
24.3
*N.B.–Average
percentage loss in 'holding' pasteurization amounted to 24.1.
1
2
3
4
Flash . . .
Do. . . .
Do.
...
Do.
...
26 4
24.9
25.5
24.6
21.8
19.8
20.7
21.3
17.4
20.5
18.8
13.4
*N.B.—The average percentage loss in ' flash ' pasteurization amounted to 17 .5.
In Table III the destruction of a significant portion of the vitamin
C content by the ' holding method ' of pasteurization can be accounted for
as due to the prolonged period of oxidation, brought about partly as a result
of exposure to air and partly to a longer contact with the metallic
surface. The lesser destruction of vitamin C in milk by the ' flash method '
of pasteurization may probably be due to a very short heating period and
lesser exposure to metals.
(iv) Effect of heating milk to the 'first boil' (Indian household method of
boiling milk) on the. vitamin C content of milk. The vitamin C content of
milk was determined before it was subjected to the ' first boil '. The milk
was heated and its ascorbic acid content was tested at the boiling point.
Thereafter, at 10, 20 and 30 minutes of boiling. The milk after being heated
was cooled under the tap and then tested for its vitamin C content.
TABLE IV
The vitamin C content of milk before and after heating
Time of boiling
0 minutes
.....
10 minutes
.....
20 minutes .....
30 minutes
.....
Vitamin C in mg. per litre of milk
Before
heating
25.8
25.8
25.2
24.6
After
heating
20.1
8.6
0.9
0.0
Per cent
loss after
boiling
22.1
66.7
96.4
100.0
Table IV shows that:
(a) The vitamin C was destroyed to the extent of about 22 per cent by
the time the first boil was attained.
(b) After 10 minutes of boiling (which is approximately the time devoted
to boiling milk in most of the Indian homes) over 66 per cent of the ascorbic
acid content was destroyed.
40
Vitamin C Content of Milk of Cows and Buffaloes
(c) After 20 minutes of boiling almost the whole of it was destroyed.
And
(d) After 30 minutes of boiling the ascorbic acid was completely des-
troyed.
(v) Effect of storage of milk on its vitamin C content. The ascorbic acid
content of fresh milk was estimated and the sample of milk then divided
into two portions and put into stoppered flasks. One portion was kept in
cold storage and was tested for its vitamin C content at intervals of 24 hours.
The other portion was kept at the room temperature and tested at intervals
of three hours until the milk curdled on heating.
TABLE V
Effect of storage on the vitamin C content of milk
(Initial vitamin C 25.2 mg.)
Vitamin C in. mg. per litre of milk
Room temperature
After 3 hours . .
After 6 hours . .
After 9 hours . .
After 11 hours . .
23.4
21.6
20.4
19.2
Cold store temperature 45°F.
After one day...
After two days . . .
After three days . . .
After four days . . .
23.4
22.2
20.4
18.6
Table V indicates that:
Milk kept at room temperature lost nearly 24 per cent of its ascorbic
acid content at the end of 11 hours and that kept in the cold store lost over
26 per cent at the end of four days.
(vi) Influence of light on the vitamin C content of milk. The ascorbic acid
content of milk was determined and one lb. of the milk was then distributed
in small glass containers. One container was exposed to direct midday sun-
light (in early June), one to diffused light and the other was kept in a dark
place. The vitamin C content of each sample was estimated at intervals of 15
minutes.
TABLE VI
Influence of light on the vitamin C content of milk
Time of
exposure
(minutes)
15
30
45
15
30
45
15
30
45
Method of exposure
Direct sunlight
Do.
Do.
Diffused light
Do.
Do.
Darkness
Do.
Do.
Vitamin C in mg.
per liter of milk
Before
exposure
24.9
24.9
24.9
24.9
24.9
24.9
24.9
24.9
24.9
After
exposure
16.8
8.9
0.0
23.5
22.2
20.8
24.6
23.9
23.5
ZAL R, KOTHAVALLA AND HARNAND SINGH GILL 41
It was found that:
(a) Vitamin C was completely destroyed when milk was exposed to
direct midday sunlight for 45 minutes.
(b) A little less than 17 per cent of it was destroyed when milk was
exposed to diffused light for the same length of time, and
(c) When milk was kept in the dark over the same period the loss in
ascorbic acid content was just a little over 5 per cent only.
(vii) Effect of the kind of metal on the vitamin C content of milk. The
vitamin C content of milk was estimated before it was subjected to the action
of various metal surfaces in the course of pasterurization. Four vessels of
the same size and capacity constructed of aluminium, nickel, tinned copper
and naked copper were obtained and to each of these 2 lb. of milk were
added. The milk was heated to 145°F. and this temperature was maintained
for 30 minutes. It was then cooled and the ascorbic acid content determined
for each vessel. Three trials were made with the ' holding method' for every
kind of metal utensil used. A similar number of trials with the same vessel
was also made with the ' flash method ' of pasteurization. The loss in the
ascorbic acid content of milk due to the action of different metals in both
the methods of pasteurization was then estimated.
TABLE VII
Action of metals on the vitamin C content of milk during pasteurization
Method of pasteurization
Holding . . . .
Flash . . . .
Holding . . . .
Flash . . . .
Holding ....
Flash ... ..
Holding....
Flash . . . .
Metal used
Nickel....
Nickel....
Aluminium....
Aluminium....
Tinned copper....
Tinned copper ....
Naked copper....
Naked copper....
Average per cent
of vitamin
C lost in
pasteuriza-
tion
8.3
5.1
26.1
21.1
28.4
23 .5
78.4
69.2
It is evident from Table VII that:
(a) The destruction of the ascorbic acid of milk was lowest in a nickel
pasteurizer and greatest in a naked copper pasteurizer.
(b) The destruction with ' holding method ' of ascorbic acid was over
78 per cent whereas in the ' flash method ' of pasteurization it was 69 per
cent with the copper surfaces.
(viii) The loss in vitamin C content of milk during its handling and pro-
cessing in a commercial dairy. This part of the work was done in order to find
out to what extent the vitamin C content of milk was affected in the normal
course of handling. and processing market milk in a dairy. As soon as the
milk was drawn from the animals its vitamin C content was determined.
Samples were taken just before pasterurization and again after the milk had
been pasteurized, cooled and bottled. Three hours after pasteurization
(which is the usual time taken to deliver milk to the customers in the rounds)
42
Vitamin C Content of Milk of Cows and Buffaloes
the vitamin C content of the unsold milk brought back to the dairy was
determined, both from half and one pound size bottles of milk.
TABLE VIII
Effect, of handling and processing milk in a dairy on its ascorbic acid content
Immediately after milking
Just before pasteurization
......
Immediately after pasteurization (flash) ...
Three hours after bottling (½ lb. bottles) ...
Three hours after bottling (1 lb. bottles) ...
Vitamin C in mg. per litre
of milk
24.8
24.3
18.4
9.5
13.6
It was found that:
In the normal process of handling, pasteurizing and bottling of milk,
in a commercial dairy the loss of vitamin C content was over 60 per cent
in half pound bottles and almost 50 per cent in one pound bottles by the
time the milk was delivered at the customer's door.
SUMMARY
The vitamin C content of the milk and the colostrum of the Sindhi
cows was found to be higher than that of the Gir, the Crossbred and the
Ayrshire cows. The Murrah buffaloes produced milk and colostrum of a
higher ascorbic acid content than the cows of most of the breeds experimented
with.
The destruction of the vitamin C content of milk was greater in the
' holding ' than in the ' flash method ' of pasteurization.
With the first boil attained, the milk lost about 23 per cent of its
ascorbic acid content, and after 10 minutes of boiling over 66 per cent and
after 20 minutes of boiling almost the whole of it.
Milk kept at room temperature lost nearly 24 per cent of its ascorbic
acid content at the end of 11 hours, while that kept in the cold store (45°F.)
lost only about 26 per cent at the end of four days.
Vitamin C was completely destroyed when milk was exposed to
direct midday sunlight (June) for 45 minutes, about 17 per cent when it was
exposed to diffused light and only 5 per cent when it was kept in the dark
for the same length of time.
The destruction of ascorbic acid was lowest when the milk was heated
in a vessel made of nickel, slightly higher in aluminium, still higher in tinned
copper and highest in naked copper. In the last case the destruction was
over 78 per cent in the ' holding' and 69 per cent in the ' flash method ' of
pasteurization.
Milk handled and processed in a commercial dairy and delivered to
customers in half pound bottles lost over 60 per cent and in one pound bottles
over 50 per cent of its vitamin C before it was delivered at the customer's
door.
ZAL R. KOTHAVALLA AND HARNAND SINGH GILL 43
ACKNOWLEDGEMENTS
The authors are grateful to the staff of the Imperial Dairy Institute,
Bangalore, for arranging and supplying the samples of milk, etc. as required.
REFERENCES
King, C. G. and Waugh, W. A. (1934). J. Dairy Sc. 17, 489
Kon, S. K. and Watson, M. B. (1936). Biochem. J. 30, 2273
Manaker, M. H. and Guerrant, N. B. (1938). Ind. Eng. Chem., Anal. Edi. 10, 25
Schwartze, E. W., Murphy, F. J. and Hann, R. M. (1930). J. Nutrition, 2, 325
Whitnah, C. H. and Riddell, W. H. (1937). J. Dairy Sci. 20, 9
PLEUROPNEUMONIA IN GOATS WITH SPECIAL
REFERENCE TO PASTEURELLA INFECTION
BY
P. G. PANDE, M.Sc., M.R.C.V.S.
Veterinary Investigation Officer, United Provinces, Lucknow
(Received for publication on 18 September 1941)
(With Plates V and VI)
P LEUROPNEUMONIA which has been described as one of the most
serious diseases affecting goats has been reported from several provinces
and has also on previous occasions been investigated at the Imperial Veterinary
Research Institute, Mukteswar. The object of this paper is to record the
results of an investigation on an outbreak of this disease in which the rô1e of
a pasteurella organism as a causative factor has been established.
HISTORY OF THE DISEASE
The first outbreak of this disease in a virulent form was reported in 1923
from Garhwal district, practically eliminating the goat population. In 1924,
outbreaks of pleuropneumonia prevailed in Almora district and accounted for
the death of 1,500 animals. Most of these outbreaks remained undiagnosed,
and no organisms of possible etiological significance were isolated. In 1926,
a form of pneumonia was investigated by Cooper in hill-goats, and in this
connection the following is quoted from Shirlaw [1939]: ' Impressed by the
large number of cases of pneumonia spontaneously occurring in goats at
Mukteswar, Cooper conducted bacteriological examination on the diseased
lungs from which he isolated an organism of the pasteurella genus which
proved pathogenic to rabbits. From an outbreak in Garhwal hills he isolated
a similar organism, thus satisfying himself that the disease of goats naturally
occurring at Mukteswar did not radically differ from that reported from the
provinces.' About the same time Haddow [Shirlaw, 1939] investigated the
disease at Izatnagar among the local goats and concluded that the condition
was an acute bacterial pneumonia, the organism involved being diplococcus,
probably a pneumococcus.
During 1937-38, a form of pneumonia in kids was investigated at the
Goat Breeding Farm, Etah, and diagnosed as bronchopneumonia. As all
these cases occurred in winter, chill was attributed to be the pre-disposing
cause. The condition was not investigated bacteriologically and its etiology
remained obscure.
An outbreak of pleuropneumonia which provided material for cultural
and experimental studies appearing in this paper occurred in August 1940,
at the Goat Breeding Farm, Etah. Six animals succumbed to the disease
out of 20 affected in a herd composed of 200 goats. The disease had pre-
vailed among the male stock, affecting only the adult bucks, and the young
44
P. G. PANDE 45
male goats and nannies kept under similar conditions had escaped infection.
The outbreak, however, subsided with the death of all the affected bucks.
The first victim of the disease at this farm fell ill on 30 August 1940, and
died on the following day ; the second was observed on 2 September 1940 and
died two days later. Of two more cases which developed on September 4,
one died on the 9th and the other on the 10th September. The last two
cases developed on September 5 and were destroyed on the 10th for the pur
pose of post-mortem examination and collection of material for further work.
SYMPTOMS
It was difficult to ascertain the incubation period of the disease in animals
in the field, but in experimental cases it varied from one to three days.
Goats of all ages were equally susceptible to experimental infections, although
cases on the farm were observed only among the adult stock. The symptoms
in both natural and experimental cases were observed to be identical, the
first sign of the disease being disinclination to eat grain and a tendency to
isolate from the main herd. The condition was in every case characterized
by extreme dullness, high temperature varying from 105° to .108° F. in the
morning and appearence of serous or thick mucoid discharge from the nose
and watery secretion from the eyes. The affected animals stood with
hanging heads and the hind legs brought forward under the belly. Sali-
vation in a few was so profuse that the ground in front was found wet with
the discharge. Respiratory symptoms were most marked. There was heavy
breathing mostly abdominal, in type. Groaning was audible at frequent
intervals and twitching of the nostrils was observed during each act of respira-
tion. At this stage the animals stood with their forelegs apart. They had
a little or no tendency to move, and if forced to move groaning at each step
could be heard. Symptoms of cough may not be noticed in acute cases of the
disease, but its occurrence has been observed in every chronic case, both under
natural and experimental attacks. Death in acutely affected animals has
been observed to supervene within 1. 2 to 24 hours from the onset of symptoms,
but at the same time a number of cases both natural and artificially infected
were also observed to continue from 4 to 6 days or more. Recovery is excep-
tional, and none of the affected animals showed diarrhoea suggestive of intes-
tinal involvement.
PATHOLOGY
Gross lesions
Autopsy on two natural cases of the disease revealed that the main
changes were confined to the lungs, the pleura and the pericardium, and varied
in extent depending upon the severity and the duration of the disease. These
post-mortem findings are summarized in detail below.
Case 571. Both the lungs were firmly adherent to the pleura on each
side ; the pleural cavity contained about 20 c.c. of slightly blood-tinged
fluid ; the apical and the middle lobes of both the lungs were found enveloped
in a thick fibrinous deposit; all the lobes of the right lung and the apical and
the cardiac lobes of the left were completely consolidated and showed
Indian
J.
Vet.
Sci.
&
Anim.
Husb.,
Vol.
XIII,
Part I] [
PLATE V
FIG. 1. The right lung showing complete consolidation of one lobe and patches of consolidation on
other lobes
FIG. 2. The cut surface of a hepatized piece of lung
showing thickening of the interlobular septa
PLATE VI
[ Indian J. Vet. Sci. & Anim. Husb., Vol. XIII, Part I
FIG. 1. Showing intense leucocytic
infiltration of the pleura and
an interlobular septum × 40
FIG. 2. Showing an early stage of hepatization commencing with the
filling up of the alveoli with sero-cellolar exudate × 40
FIG. 3. Showing a patent bronchiole and throm-
bosis of bronchial vessels Section
prepared from an area of pulmonary
infarct, and a scrutiny will reveal
congestion of the alveolar capillaries × 40