NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
December 1948]
NOSHIR
N. DASTUR,
et
al.
237
The results show that both the vegetable and animal rennets are sensitive to heat. Heating
over 50°C. even for short duration, effects the activity of the solutions of these rennets adversely.
(iv) Effect of using boiled milk. Milk was brought to first boil and then allowed to cool to 37°C.
As substrates milk and milk containing CaCl 2 were used. The results are given in Table V.
TABLE V
Effect of using boiled milk on the activity of Withania coagulans enzyme
Milk used
Cow milk raw
.
.
.
.
.
.
.
.
.
.
.
Cow milk boiled
.
.
.
.
.
.
.
.
.
.
.
Cow milk boiled-fortified with
Ca.
.
.
.
.
.
.
.
.
.
.
Buffalo milk
raw.
.
.
.
.
.
.
.
.
.
.
Buffalo milk
boiled.
.
.
.
.
.
.
.
.
.
.
Buffalo milk boiled-fortified with Ca.
.
.
.
.
.
.
.
.
.
.
10 per cent
animal rennet
5 per cent
vegetable rennet
in CaCl 2
(Coagulation time in seconds)
30
39
. .
26
32
27
70
97
81
47
74
62
It was found that heated milk takes longer time to coagulate. Action of vegetable rennet was
much slower than that of animal rennet. Addition of CaCl 2 compensates to a large measure the
effect of heat. The figures for the relative time for the coagulation of cow and buffalo milk are
interesting. Buffalo milk on an average was found to take less time for coagulation either with
animal or vegetable rennets.
(v) Effect of various absorbents. A 5 per cent aqueous solution of acetone precipitated enzyme
was used. 50 ml. of this solution were treated at room temperature with 5 per cent of the follow-
ing :–
(a) Animal charcoal (++++++).
(b) Rice husk charcoal (+++++).
(c) Fuller's earth (++++++++).
(d) Kaolin (+).
(e) Aluminium oxide (++++).
(f) Calcium carbonate (+++).
(g) Calcium hydroxide (++++++++).
(h) Kiesulghar (++).
After keeping for about 30 minutes with occasional stirring, the solutions were filtered through
a filter paper. The degree of clarification obtained was noted visually and is indicated by (+) signs.
Activities of the filtrates were tested using milk acidified to 0.2 per cent lactic acid and fortified
with CaCl 2 . The results are shown in Table VI.
238
Preparation of Vegetable Rennet
[Vol. XVIII, Part IV
TABLE VI
Activity of vegetable rennet solutions treated with absorbants
Original
solution
149
Vegetable rennet solution treated with 5 per cent of
Animal
charcoal
231
Rice husk
charcoal
152
Fuller's
earth
305
Kaolin
Aluminium
oxide
Calcium
carbonate
(Coagulation time in seconds)
243
166
160
Calcium
hydroxide
Over 1,200
Kiesulghar
229
It was found that Ca(OH) 2 wholly destroyed the activity of the filtrate. Similar results were
obtained even with 1 per cent Ca(OH) 2 . Fuller's earth, Kiesulghar, Kaolin and animal charcoal,
calcium carbonate and aluminium oxide and rice husk charcoal decreased the activity in the order
mentioned. Attempts were made to extract the enzyme from these absorbents with various media
without success. Some of the absorbents were alkaline and so the filtrate obtained was acidified
and then the activity tested. Still the solutions did not show enhanced activity.
(vi) Toxicity of Withania coagulans extract. This was tested with rats. Four young male
rats were fed 4 ml. of a 10 per cent extract of the berries for a period of ten weeks. An equal number
of animals were kept as controls. The weight of animals was recorded. No difference was found
in the growth rate of control and experimental groups of animals. It was concluded that the extract
of the berries was non-toxic.
(vii) Effect of different salts and sunlight. That calcium chloride considerably enhances the
activity of the Withania coagulans enzyme has been indicated previously. Effect of other inorganic
salts was investigated. Amongst the salts tried were ZnCl 2 (0.004 per cent), KI (0.005 per cent).
Cu 2 O (0.04 per cent) and bleaching powder (0.04 per cent). The salt to be tested was added to milk.
Vegetable rennet solution was made in 12.33 per cent CaCl 2 . The results are given in Table VII.
TABLE VII
Effect of salts on the activity of vegetable rennet
5 per cent
Vegetable rennet
in water
210
5 per cent vegetable rennet in 12.33 per cent CaCl 2 solution
—
78
ZnCl 2
(0.004 per cent)
194
Salt added to milk
Cu 2 O
(0.04 per cent)
KI
(0.005 per cent)
Coagulation time in seconds
107
84
Bleaching powder
(0.04 per cent)
88
It is seen that none of the substances tried except CaCl 2 enhanced the activity. In fact they all
produced a slight inhibitory effect.
Vegetable rennet solution in H 2 O and in CaCl 2 were exposed to strong sunlight and the activity
determined at intervals. Strong sunlight was found not to effect the activity of vegetable rennet.
Narain and Singh [1942] also state that radiations of different wave-lengths have no influence on
the activity of the enzyme.
December 1948]
NOSHIR
N. DASTUR, et al
239
(viii) Preparation of cheese. Both soft (Surti panir) and hard (Chaddar) cheeses were prepared
from various concentrates of Withania coagulans obtained from time to time. As promising results
were obtained in the beginning with the vegetable rennet, no attempt was made to deviate from the
standard technique followed with animal rennet. A few difficulties were encountered in the begin-
ning but these were gradually overcome. It will, however, be interesting to record some of the main
defects encountered in the beginning for information.
(a) Soft cheese. The texture was very crumbly. Cheese when kept dipped in whey gradually
became thinned at the edges.
(b) Hard cheese. Cheese developed open texture on keeping. There was a bitter and biting
taste which made the product unpalatable. Cheese was soft and would not cut clean.
As better quality rennet material was obtained, these difficulties were gradually overcome and
it was possible to make as good a soft cheese with vegetable rennet as with the product obtained
from animal source. A summary of a comparative operation for the manufacture of Surti panir is
shown in Table VIII.
The method of preparation of Surti panir has been described in detail by Kothavalla and Verma
[1942] and was followed in the present investigation. For dipping vegetable rennet cheese, various
media, including animal rennet whey, 1.5 per cent salt solution, 0.05 per cent lactic acid solution in
1.5 per cent NaCl, were tried and in the end it was found that vegetable rennet whey obtained during
the operation was the most satisfactory.
TABLE VIII
Preparation of Surti panir with animal and vegetable rennets
Particulars of operation
Milk taken (6.5 per cent fat)
.
.
.
.
.
.
.
Temperature of
milk.
.
.
.
.
.
.
Starter added
.
.
.
.
.
.
.
.
Acidity of starter
.
.
.
.
.
.
.
Amount of rennet
added.
.
.
.
.
.
.
Time rennet added
.
.
.
.
.
.
.
Time cut
.
.
.
.
.
.
.
Time first turned
.
.
.
.
.
.
.
Time dipped in whey
.
.
.
.
.
.
.
Fat per cent of whey
.
.
.
.
.
.
.
Remarks: Colour
.
.
.
.
.
.
.
Texture
.
.
.
.
.
.
.
Taste
.
.
.
.
.
.
.
Hansen's liquid rennet
2 lbs. . . . .
95°F
.
.
.
.
1 oz . . . .
0.65 per cent lactic . .
4 drops
.
.
.
.
9-30 a.m. . . . .
10-40 a.m. . . . .
11-45 a.m. . . . .
4-30 p.m. . . . .
0.50 per cent . . . .
White
.
.
.
.
Good
.
.
.
.
Good
.
.
.
.
Vegetable rennet
5 per cent Soln. in
12.33 per cent CaCl 2
2 lbs.
95°F.
1 oz.
0.65 per cent lactic.
24 drops.
9-35 a.m.
10-45 a.m.
11-50 a.m.
4-30 p.m.
0.50 per cent.
White.
Good.
Good.
On the whole Surti panir prepared by animal and vegetable rennets were indistinguishable.
Trials were also carried out to prepare Chaddar cheese with vegetable rennet. By trials it was
observed that longer time is required for the ripening of Chadder cheese made with Withania
coagulans and it should possibly be extended to six months. As the time of ripening increased, the
bitter taste diminished. It has, however, not been possible to remedy this defect completely.
Further work is in progress to purify the enzyme and it is hoped that it will be then possible to get
over this difficulty.
240
Preparation of Vegetable Rennet
[Vol. XVIII, Part IV
SUMMARY
1. Seeds, leaves and stems of Withania coagulans show no milk clotting property. The pulp o
the berries was the most active source of the enzyme. A little activity was also shown by the outer
husk.
2. For the preparation of an enzyme concentrate, the enzyme was first extracted with wate
and then precipitated with two volumes of acetone. The activity of the acetone precipitated enzym
compared well with that of Hansen's liquid and powder rennets.
3. The optimum temperature of Withania coagulans enzyme lay between 45-65°C. At higher
temperatures than 70°C., the activity of the enzyme decreased rapidly and it was completely inactive
at 80°C.
4. The activity of the enzyme increased with an increase in the activity of milk.
5. Boiled milk took a longer time to clot but the lost activity could be restored to a large measure
by adding calcium chloride.
6. Withania coagulans extract is non-toxic.
7. Different absorbents decrease the activity to different degrees.
8. Soft cheese of a quality indistinguishable from that made with animal rennet was prepared.
ACKNOWLEDGMENTS
The authors wish to express their thanks to the Indian Council of Agricultural Research for
giving a special grant for this work. They are also grateful to Dr K. C. Sen, Director of Dairy
Research and Dr Zal R. Kothavalla, Dairy Development Adviser for their helpful suggestions and
criticism.
REFERENCES
Kothavalla, Z. R. and Khubchandani, P. G. (1940). Ind. J. vet. Sci., 10, 284
——and
Verma, H. C. (1942). Ibid., 12, 297
Lea, S. (1883-84). Proc. Roy. Soc., London, 36, 55
Narain, R. and Singh, A. (1942). Ind. J. ret. Sci., 12, 224
UTILIZATION OF WHEY—A MILK WASTE IN THE PRODUCTION OF
'CHHANA'-FOR SUPPLY OF CALCIUM TO THE POOR RICE DIET
BY H. N. DE AND A. B. SOM. From the Nutrition Research Unit, I.R.F.A., Biochemical Laboratory,
University of Dacca, Dacca
(Received for publication on 14 May 1948)
C ALCIUM deficiency is one of the fundamental defects of the poor rice-eaters' diet in India and
attempts have been made in this laboratory to explore some cheaper sources of calcium to be
supplemented with the poor rice diet Basu et al [1942]. It has been shown, that, the lime taken up
in the process of chewing betel leaves and bones of small fish, can make up the calcium deficiency
of the poor rice diet. Further attempts have been made to find out some other cheaper source
of nutritionally available calcium.
which is employed in the manufacture of sweets consumed by the richer section of the population.
The whole amount of whey containing valuable nutritive materials as lactose, calcium, protein etc.
is thrown away every day as waste. In a province like Bengal about 34,000 md., i.e., 1,200 tons of
whey is thrown away daily as such. Our aim was, to see whether this whey which is thrown away
as waste can be utilized as food supplement for supply of calcium to the poor rice-eaters' diet of
India.
EXPERIMENT AND RESULTS
Metabolic experiment was carried out on two healthy adult human subjects of 42 and 44 kg.
body weight and 11 adult rats of 162 to 213 gms. body weight who were kept on the poor rice diet
of the following composition.
TABLE I
Composition of the diet
Name of the foodstuff
Rice . . . . . . . . . . . . . . . .
Pulse. . . . . . . . . . . . . . . .
Vegetables :—
Potato. . . . . . . . . . . . . . . .
Green banana. . . . . . . . . . . . . .
Cabbage.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
Oil-Groundnut . . . . . . . . . . . . . . .
Weight in
grammes
550
100
100
50
50
30
Daily whey supplement
(a) For human subject -200 c.c.
(b) For rats -6 c.c.
In case of the human experiments, the above quantity of the diet was given daily to the subjects
and in case of rat experiments, the basal diet was prepared according to the above composition after
drying all the ingredients and each rat was given 10 to 15 gms. of the diet according to the require-
ment. Whey supplementation was begun after a basal period of six days on the rice diet and this
was continued for further period of six days. In both basal and supplementation periods the collec-
tion of urine and faeces was made on the last three days of the period and their calcium was estimated
according to the method adopted in the previous communication from this laboratory [Basu et al
1942].
241
I
242
Utilization of Whey
[Vol. XVIII, Part IV
In case of human experiments, the whey supplement was administered along with 'dal' and
vegetables after cooking them with it, but in case of rat experiment, the whole diet was cooked with
whey. Whey was daily collected from the market and its calcium content was found to range between
0.62 to 1.3 mg. per c.c. The results of metabolic experiments with human subject and rats are
presented in Table II from which it is observed that both the subjects J. C. D. and K. N. S. maintained
negative calcium balance when living on poor vegetarian rice diet. The average intake and balance
of calcium on rice diet was found to be 237.1 mgm. and—74.3 mgm. respectively. The administration.
of 200 c.c. of whey daily to the diet furnished on average 133.4 mgm. of extra calcium to the diet and
improved the negative balances of the two subjects J. C. D. and K. N. S. from —127.3 to +4.4 mgm.
and from —21.2 to +83.3 mgm. respectively. The percentage utilization of extra calcium of whey
supplement was calculated to be 94.5 and 82.5 respectively.
In adult rat experiments the mean daily calcium intake and balance were found to be 9.01 and
—0.7 mgm. respectively and the supplementation of 6 c.c. of whey to the diet supplied on average
daily 5.73 mgm. of extra calcium and improved the balance from —0.7 to +4.1mgm. In this case
the average percentage utilization was found to be 83.7.
Although the whey is produced as waste from milk, the utilization value of its calcium determined
in the present work, is found to be considerably higher than that of milk. By a series of balance
studies on children and adults Kinsman et al [1939], Steggerda and Mitchell [1939] and Brieter et
al [1941] have observed the utilization value of milk and calcium between 12 to 29 per cent.
TABLE II
The calcium utilization of whey by adult human subjects and rats. Each period consisted of
three days and the figures indicate the daily averages for the three days collection. The periods
which are not indicated in the Table should be regarded as preliminary periods
Human experiment
Experimental subject
J.C.D.
Body wt. -42 Kg.
Age -22 years
K.N.S.
Body wt. -46 Kg.
Age -25 years
Average body wt. of the
rats -
187 gm.
Experi-
mental diet
Rice diet
Rice diet
Rice diet
Rice diet
Rice diet
Rice diet
Supplement
Nil
200 c.c.
whey
Nil
200 c.c.
whey
Period
II
IV
II
IV
Dietary
calcium
mg. per day
248
248+
139.3
=387.3
226.2
226.2+
127.5
= 353.7
Rat experiment
Nil
6 c.c.
whey
II
IV
9.01
9.01 +
5.73
= 14.74
Total out-
put of
calcium
(urinary and
faecal) mg.
per day
375.3
382.9
247.4
269.9
9.71
10.61
Balance
of calcium
mg. per day
—127.3
+4.4
—21.2
+ 83.8
—0.7
+ 4.1
Per cent
utilization
of whey
calcium
––
94.5
––
82.5
—
83.7
December 1948]
H. N. DE AND A. B.
SOM
243
The higher utilization value of whey calcium above 80 per cent observed here in both human
and rat experiment may be due to the presence of sufficient quantity of lactic acid in it.
It has previously been pointed out that about 1,200 tons of whey is daily thrown away as waste
in Bengal. Taking the average calcium content of whey as 0.7 mg. per c.c., the net utilizable calcium
wasted away daily along with whey is approximately 9 tons. If 200 c.c. of whey is given as food
supplement to each adult, the above quantity of whey can make up the deficiency of 5.5 million
people of Bengal. So for practical nutrition, whey thrown away as waste in the production of sweets
for rich section of the people may be used as a calcium supplement for the poor and the present work
opens a new avenue for the fortification of poor Indian diets.
SUMMARY
Administration of 200 c.c. of whey with the poor rice diet would supply about 137 mg. of extra
calcium and make up the deficiency of the rice diet with regard to this essential element. By both,
rat and human experiments the percentage utilization of the whey, calcium was found between 80
to 90 per cent.
REFERENCES
Basu, K. P., Basak, M. N. and De, H. N. (1942). Ind. Jour, Med. Res., 30, 309
Basu, K. P., De, H. N. and Basak, M. N. (1942). Ibid 30, 417
Kinsman, G., Shaldon, D. Jensen E., Bernds. M., Outhouse, J. and Mitchell, H. H. (1939). J. Nutrition 17, 429
Steggerda, F. & Mitchell. H. H. (1939). Ibid. 17, 253
Idem (1941).—Ibid 81, 577
Breiter, H., Hills. R., Dwight, J., McKey, B., Armstrong, W. and Outhouse, J. (1941). 21, 417
246
Khakhra Dhand in the Punjab
[Vol. XVIII, Part IV
Khakhra tributary in which, the Khakhra 'Dhand' lies, is the old bed of the river Beas. The
rivers Sutlej and Beas at one time flowed parallel to each other for a considerable distance in the
districts of Amritsar, Lahore and Montgomery. As the river Beas now falls into river Sutlej at
' Hari-ke-Pattan', about twenty miles above the confluence of the channel of the Khakhra tributary
into the river Sutlej, the Khakhra tributary serves as a drain of the low-lying area between the river
Sutlei and the old bed of the river Beas.
The level of the water in the Khakhra 'dhand' and the channel of the Khakhra tributary rises,
as the level of the water in the river Sutlej gets higher due to the melting of snow, ultimately flooding
the fields lying on either side of the channel and some part of the 'dhand'.
Apart from the Khakhra 'dhand', a few more big pools are found in the Khakhra tributary
in its upper reaches. But, they are not appreciably affected by the rise of level in the river Sutlej
during the floods.
FIG. 1. Khakhra Dhand the spawning ground for Catla and other carps
Evidence of Catla breeding in the 'Dhand'
The actual courtship, wed-lock and spawning have not been observed, but the following facts
indicate fairly clearly that the fish breeds in the 'dhand'.
1. Large-sized Catla, presumably sexually mature, have been observed entering the 'Dhands'
from the river Sutlej in the month of June.
2. Spent Catla have been caught while going down to the river in September.
3. Sixty per cent of the fish population of the 'dhand' consists of young Catla.
4. Flooding of the river and in consequence of the channel and the 'dhand' takes place in June
and July owing to the melting of the snow, but independent of monsoon rains. As floods due to
melting of snow occur every year, the spawning also occurs in the 'Dhand' every year.
5. The sides of the channel as well as of the 'dhand' provide suitable spawning beds when covered
with flood water.
6. The temperature never rises above 90°F., and is therefore, suitable for Catla spawning.
December 1948]
UDE
SINGH
RAI
247
Composition of 'Dhand' water before and during flooding
TABLE I
Results of water-analysis of Khahhra 'dhand' per 100,000 parts, during June and July, 1946
Date
12-6-46
27-6.48
29-7-46
28-7-46
Place of
collection
Surface
bank
Bottom
bank
Surface
middle
Bottom
middle
Surface
Bank
Bottom
bank
Surface
middle
Bottom
middle
Surface
bank
Bottom
bank
Surface
middle
Bottom
middle
Surface
bank
Bottom
middle
Surface
middle
Bottom
middle
Total
salts
204.25
207.63
212.73
220.52
41.37
36.89
29.30
31.29
—
—
—
31.89
31.95
30.25
31.25
Car-
bona-
tes
23.32
20.14
19.08
21.2
2.12
8.48
6.36
12.72
—
—
—
Nil
Nil
Nil
Nil
Bi-car-
bonates
152.04
157.08
156.24
168.00
20.16
18.45
11.70
10.08
—
—
—
21.84
17.64
15.12
16.18
Chlo-
rides
22.8
23.4
21.6
28.08
3.51
3.51
5.85
5.85
—
—
—
5.26
4.12
4.68
5.26
Sul-
phat-
es
6.27
7.98
16.7
9.38
13.4
9.2
5.17
4.87
—
—
—
5.79
10.46
10.15
9.15
Cal-
cium
Traces
Traces
Traces
Traces
2.96
1.92
0.96
1.68
—
—
1.56
1.8
1.48
1.56
Dissolved
oxygen per
litre
2.657 cc.
2.279 cc.
2.519 cc.
2.183 cc.
4.100 cc.
3.543 cc.
3.768 cc.
3.457 cc.
3.756 cc.
3.617 cc.
3.798 cc.
3.788 cc.
3.150 cc.
3.045 cc.
3.642 cc.
3.349 cc.
Required
oxygen
3.59 gms.
3.66 gms.
3.16 gms.
2.92 gms.
0.315 gms.
0.825 gms.
0.230 gms.
0.280 gms.
0.660 gms.
0.640 gms.
0.720 gms.
0.70 gms.
0.515 gms.
0.55 gms.
0.58 gms.
0.44 gms.
pH
8.36
8.38
8.47
8.47
9.96
9.94
9.95
9.93
7.0
7.0
7.0
7.0
7.0
7.0
7.0
7.0
Phos-
phates
Traces
Traces
Traces
Traces
Traces
Traces
Traces
Traces
—
—
—
—
Traces
Traces
Traces
Traces
Nitra-
tes
0.0625
mgms.
0.04
mgms.
0.0125
mgms.
0.015
mgms.
0.0325
mgms.
0.0375
mgms.
0.02
mgms.
0.0325
mgms.
—
—
—
—
0.126
mgms.
0.124
mgms.
0.125
mgms.
0.126
mgms.
248
Khakhra
Dhand in the Punjab
Vol. XVIII, Part IV
TABLE II
Variation in amount of total salts in 'Khakhra dhand' water, from October, 1945 to September 1946
Month
October, 1945 . . .
November, 1945. . .
December, 1945 . . .
January, 1946 . . .
February, 1946 . . .
March, 1946. . .
April, 1946 . . .
May, 1946
.
.
.
12 June, 1946 . . .
27 June, 1946 . . .
28 July, 1946 . . .
31 July, 1946 . . .
10 August, 1946 . . .
21 August, 1946. . .
31 August, 1946 . . .
11 September, 1946 . . .
27 September, 1946 . . .
Total salts per 100,000 parts
67.71 parts to 97.25 parts
78.20 parts to 100.2 parts
81.05 parts to 83.2 parts
82.22 parts to 94.20 parts
88.42 parts to 89.25 parts
100.45 parts to 101.23 parts
127.92 parts to 170.25 parts
145.05 parts to 174.27 parts
204.25 parts to 220.52 parts
29.30 parts to 41.37 parts
30.25 parts to 31.95 parts
29.79 parts to 31.05 parts
28.12 parts to 29.09 parts
75.26 parts to 78.40 parts
71.01 parts to 76.18 parts
88.56 parts to 90.25 parts
114.28 parts to 115.25 parts
From the Table above it is observed that—1. The presence of total salts in the 'dhand' water
continued to increase from October, 1945 to June, 1946. Ultimately in June 1946, the increase
of total salts in the 'dhand' water was found to be ten times to that in the river water, as is evident
from the Table II, given above.
2. River-water entered the 'dhand' during the second fort night of June, 1946 and diluted the
'dhand' water, as is clear from the figures given in Table II.
3. Analysis of 'dhand' water revealed that its pH value was 9.9 before the spawning, and the
pH value was 7.0 after the spawning had taken place due to the dilution caused by the inrush of
river-water into the 'dhand', as is evident from the data in Table I.
The increase of total salts, which are mostly 'kallar' salts, namely—carbonates, bicarbonates,
chlorides and sulphates of sodium and potassium present in the 'dhand' water, is due to the addition
of these salts from the seepage-water received by the 'dhand' water during these months. It is to
be noted particularly that there were no rains from October 1945 to June, 1946, which could wash
into the 'dhand' the ' kallar' salts lying in large quantity in the fields on either side of the' dhand'.
Significance of the composition of water in the behaviour of fish
It has been found out that the 'dhand' contains no adult Catla from October to June. The
adult fish in the river are attracted into this 'dhand' due probably to the presence of brackish water
in the 'dhand', when the level of river-water rises in June-July. They lay their eggs and then leave