NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
S. N. RAY 211
The cause for these widely divergent figures quoted for magnesium require-
ment is probably due firstly to the differences in the requirements of various
breeds and types of animals used by the different workers and secondly to the
facts that the availability of the magnesium present in different food-stuffs
may vary considerably and that the apparent magnesium content of the feed
may not be any guide to the amount of magnesium which will be utilized by
the animal. This theory is supported by the findings of Carbery, Chatterjee
and Talapatra [1937], who found a positive retention of magnesium using
'Aman' paddy straw as a single feed, whereas the administration of 'Aus'
paddy straw alone resulted in a deficiency of the mineral in most of the animals,
even though the latter feed containd a greater concentration of magnesium.
The non-availability of other minerals in the feed is well known. Thus Elvejhem,
Hart and Sherman [1933] found the iron in vegetable and animal material
available for haemoglobin formation to be only a fraction of the total iron
present. Similarly phytin, found in various cereals, is a compound which
animals are unable to assimilate. Consequently the phosphorus it contains
[Bruce and Callow, 1934] is not available to them. Phytin also contains
magnesium and consequently the magnesium present in this form is also lost
to the animals. The total quantity of magnesium present as phytin in the
feed is, however, comparatively small and hence the content of phytin alone
cannot be the sole determinant of the non-availability of this mineral in the
ration. If the negative balance of magnesium in animals on feeds such as
those used in these experiments is to be explained on the theory of the non-
availability of some of the magnesium in the feed, it will be necessary to seek
for other compounds besides phytin from which magnesium is not available.
SUMMARY
The magnesium content of the blood sera of a number of animals suffering
from fainting ' fits ' was found to be low. The calcium concentrations were
found to be normal.
The magnesium concentrations were found to diminish during the fainting
periods and to increase immediately after the ' fit'.
The symptoms in one of the animals were found to disappear on changing
its roughage from wheat-straw to fresh hay.
The administration of magnesium carbonate increased the magnesium
content of the serum but did not relieve the symptoms of the malady.
Metabolism experiments were conducted on four animals and three of
them were found to be on a negative balance with reference to magnesium.
On the particular feeds used, 24 mg. of MgO per lb. of body weight was found
to be just about sufficient to keep the animals in magnesium equilibrium.
ACKNOWLEDGEMENTS
My sincere thanks are due to Mr J. R. Haddow, Officer-in-charge, Patho-
logy and Bacteriology Section and to Dr K. C. Sen, Officer-in-charge, Animal
Nutrition Section for their advice and encouragement. My thanks are also
due to Mr K. Govind Rau for his technical assistance and many careful
analyses.
D
212
Hypomagnesaemia
in Heifer Calves
REFERENCES
Allcroft, W. M. and Green, H. H. (1938). J. Comp. Path. & Therap. 51, 176
Bruce, H. M. and Callow, R. K. (1934). Biochem. J. 28, 517
Carbery, M., Chatterjee, I. and Talapatra, S. K. (1937). Indian J. Vet. Sci. & Anim.
Husb. 7, 155
Dukes, H. H. (1937). The physiology of farm animals. (London) 4th revised edition,
p. 48
Duncan, C. W., Huffman, C. F. and Robinson, C. S. (1935). J. Dairy Sci. 19, 440
Duncan, C. W., Lightfoot, C. C. and Huffman, C. F. (1940). J. Dairy Sci. 23, 125
Elvejhem, C. A., Hart, E. B. and Sherman, W. C. (1933). J. Biol. Chem. 103, 61
Fiske, C. H. and Subbarow, Y. (1925). J. Biol. Chem. 66, 375
Forbes, E. B., French, R. B. and Letonoff, T. V. (1929). J. Nutrition 1, 201
Greenberg, D. M. and Tufts, E. V. (1935). Proc. Soc. Expt. Biol. & Med. 32, 674
Hirschfelder, A. D. (1933). Proc. Soc. Expt. Bipl. & Med. 30, 996
Huffman, C. F. and Duncan, C. W. (1936). J. Dairy Sci. 19, 440
Huffman, C. F. and Robinson, C. S. (1926). J. Biol. Chem. 69, 101
Kemmerer, A. R., Elvehjem, C. A., Hart. E. B. and Fargo, J. M. (1932). Amer. J.
Physiol. 102, 319
Kramer, B. and Tisdall, F. F. (1921). J. Biol. Chem. 47, 475
Kruse, H. D., Orent, E. R. and McCollum, E. V. (1932). J. Biol. Chem. 96, 519
McNeille, L. G. (1934). J. Amer. Med. Assoc. 103, 548
Orent, E. R., Kruse, H. D. and McCollum, E. V. (1932). Amer. J. Physiol. 101, 454
Seekles, L., Sjollema, B. and Van der Kaay, F. C. (1931). Biochem. Zeit. 243, 316
Sjollema, B. (1930). Vet. Rec. 10, 425
—(1932).
Nutr. Abs. and Rev. 1, 621
Sjollema, B. and Seekles, L. (1930). Biochem. Zeit. 229, 358
Talapatra, S. K., Ray, S. C. and Sen, K. C. (1940). Indian. J. Vet. Sci. & Anim.
Husb. 10, 243
Wendt (1925), Handbuch der Biochemie, Edited by C. Oppenheimer, 2nd edn. vol. 8.
183
USE OF SELECTED STARTERS IN THE PREPARATION
OF DAHI
BY
M. R. MADHOK, PH.D., ASSOC. I.A.R.I.
AND
S. L. KAPOOR, M.Sc.
Punjab Agricultural College, Lyallpur
(Received for publication on 13 September 1940)
T HE importance and application of selected starters in the ripening of
cream, in the preparation of cheese and allied products, are too well
known to need emphasis, but in so far as their use in the preparation
of dahi is concerned, their utility has not been duly appreciated in India.
A greater part of the total milk supply in India is consumed as curd
or soured milk (dahi) and unlike the practice in most other countries, this
product is churned for butter. Curd with poor texture and bad flavour
is not acceptable because of its poor palatability and lower butter yield.*
Work done in India connected with milk souring is that of Ram Ayyar
[1928], which describes the presence in dahi of different types of lactic acid-
producing organisms during different seasons, and that of Joshi and Ram
Ayyar [1935] and Karnad [1939] on the aroma-producing organisms in dahi.
The present work describes investigations on the effect of some pure culture
starters alone and in combination on the quality of dahi.
EXPERIMENTAL
Isolation of organisms
The lactic acid-producing organisms were isolated from several curd
samples collected from the bazaar and houses. These were plated in loop
dilutions on purple lactose agar of the following composition :—
Beef extract . . . . . .
Peptone . . . . . . .
Lactose . . . . . . .
Agar agar . . . . . . .
Water . . . . . . .
Reaction . . . . . . .
3 gm.
5 gm.
10 gm.
15 gm.
1000 c.c.
pH 0 .8 with Brom cresol purple
as an internal indicator.
After an incubation of two days the acid-producing colonies were picked
out and transferred to sterilized skim milk in tubes. These cultures were
repeatedly sub-cultured.
* Trials conducted by us have shown that unripe or over-ripe curd, gives a lesser
butter yield by churning with the indigenous method.
213
D2
214
Starters in the Preparation of Dahi
More than a dozen cultures of acid formers were initially isolated and
after a study of their morphological characteristics, three selected forms
were identified by their cultural and physiological behaviour [Bergey, 1939]
as Streptococcus lactis, Streptococcus cremoris and Lactobacillus acidophilus.*
Since yeasts are found commonly associated with lactic organisms in samples
of dahi, a type of yeast (Saccharomyces elipsoides) was also isolated to study
its possible effect on the production of aroma and flavour in curd.
Trials
To test their efficiency in dahi production, the pure cultures were in-
oculated both singly and in different combinations into 20 ml. of cold simmered
or sterilized milk in conical flasks.
Since curd in an Indian house-hold is generally made after simmering
the milk, this investigation was carried out in simmered full cream cow's
and buffalo's milk. But in order to eliminate the influence of any chance
contaminant in simmered milk and to make the conditions technically perfect
for the study of pure cultures, a simultaneous study was also conducted in
sterilized milk samples. Simmering was done on a low flame for 1½ hours
and 20 ml. portions of the simmered milk transferred to sterilized conical
flasks. Sterilization of the milk was done at 10 lb. pressure for 10 minutes
on two consecutive days in 100 ml. conical flasks, each containing 20 ml.
milk.
Since 37°C. is the optimum growth temperature for L. acidophilus and
30°C. for the other types, combinations in which L. acidophilus formed one
of the types were incubated at both temperatures. The following is the list
of the various combinations tried:—
Temperature of incubation
30°C.
.
.
.
.
.
37°C.
.
.
.
.
.
Species in the cultures †
L, C, LY, CL, CY, AL, AC, LCY, CAL, AYL, AYC,
LCAY
A, AY, AL, AC, CAL, AYL, AYC, LCAY
Methods of comparison
1. The titratable acidity of the samples was determined at intervals of
12, 18 and 24 hours by titrating a 20 ml. sample against standard NaOH
using phenolphthalein as indicator. The acidity was reported as percentage of
lactic acid.
* L. acidophilus was isolated from a curd sample prepared with acidophilus
culture obtained from the Pasteur Institute, Kasauli.
† L=S. lactis C—S. cremoris
A=L. acidophilus
Y— Yeast
M. R. MADHOK AND S. L. KAPOOR 215
2. The taste, aroma and flavour of the curds were judged with the kind
cooperation of two experienced members of the Department of Agriculture.
3. Determinations of volatile acidity were made after 18 hours' incuba-
tion by the method used at the Government laboratory [Richmond, 1930].
RESULTS
The results showing the development of acidity, rate of curdling and the
amount of volatile acidity developed in different cases are given in Tables
I—III.
Development of acidity
Table I shows that the acidity was rather low at the beginning but in-
creased considerably in 24 hours. Since the acitivity varied considerably
with different cultures, the results for pure and mixed cultures will have to be
discussed separately.
(i) Pure cultures.—The individual cultures varied greatly in their capacity
to develop acidity. Within the scope of the present experiments A was
found to develop higher acidity than L or C. This property was
found to hold for all the different types of milk used in the experiments.
(ii) Mixed cultures.—Since great differences were observed in the indi-
vidual acid-producing capacity of the three cultures, their associated action
was expected to yield significant results.
(a) Combinations of two types.—C in pure culture developed the
lowest acidity but mixed with L it gave higher acidity than C and
slightly lower than L alone. But in sterilized milk, L and C
developed a higher acidity than either L or C individually.
Similarly A + C developed a higher acidity than C and lower than
A.
The association of yeast with bacterial cultures gave varying results.
With yeast in combination with A the acidity developed in 12 hours
was higher and subsequently lower than by the pure culture, A. In
sterilized milk the increase continued up to 18 hours. The association of
yeast with C developed lower acidity than C.
(b) Combinations of three types.—A mixed culture of A+Y+C developed
higher acidity in simmered milk than A+C or C+Y, but lower than that of
A+Y (Table I). The same results were not given in sterilized milk. The
developed acidity was lower than A+C up to 18 hours and throughout the
incubation period than A+Y. The acidity with A+Y+C was throughout
higher than with C+Y.
Rates of curdling
The results given in Table II show that the time taken to curdle milk
varied with the species in the cultures. No direct relation could be observed
between the acid producing power of A in pure culture or in combination
with other organisms and its curdling property ; thus A gave high values
or acidity but took 36 hours for curdling.
216
Starters in the Preparation of Dahi
Similarly A+Y+C developed a higher acidity than L but took a
longer time to curdle (Tables I and II).
In general it was observed that under similar conditions, relatively
longer periods were required for curdling simmered than sterilized milk.
Quality of curd
To determine the relative quality of the curd produced in different samples
the grading of flavour, aroma, appearance, compactness and the texture
of the different types was undertaken with the help of experts. For com-
parative purposes a few samples from the bazaar and houses were also ex-
amined.
Although all the experimental products scored higher than bazaar and
house samples a few were found to be much superior. These were graded
into three classes in descending order as follows :—
Class I
.
.
.
.
.
.
.
Class II
.
.
.
.
.
.
.
Class III
.
.
.
.
.
.
.
C, A, A+Y
C+Y, A+Y + C
C+L, A + C
It was found that C alone and in combination with other organisms pro-
duced a sweet curd while L and A gave a sour curd. In general,
a close parallelism existed between the taste and acidity of different curds.
Determinations of volatile acidity
Determinations of volatile acidity were clone in order to qualify to a
certain extent the personal factor in grading the product by arbitrary stand-
ards of smell and taste. C and A generally produced the largest
amount of volatile acids (Table III).
SUMMARY
The use of pure cultures alone and in combination for the preparation of
dahi has been investigated.
Four organisms, viz. S. lactis, S. cremoris, L. acidophilus and a yeast
(Sacch. elipsoides) were isolated and various combinations were used for the
preparation of dahi. Titratable and volatile acidity were determined, and
curdling capacity, aroma and taste judged in order to evaluate the quality
of the product of each culture. Seven combinations were selected for their
marked superiority ; these were placed in three classes as follows :—
(1) C,A, A+Y
(2) C+Y, A+Y+C
(3) C+L, A+C
REFERENCES
Bergey, D. H., et al. (1939). Bergey's Manual of Determinative Bacteriology (Bailliere
Tindall and Cox, London)
Joshi, N. V. and Ram Ayyar, C. S. (1936). Indian J. Vet. Sci. & Anim. Husb. 6, 141
Karnad, R. (1939). Indian .J. Vet. Sci. & Anim. Husb. 9, 349
Ram Ayyar, C S. (1928). Agric. Jour. Ind., 23, 107
Richmond, H. D. (1930). Dairy Chemistry. p. 193, (Griffin and Co., Ltd., London)
M. R. MADHOK AND S. L. KAPOOR 217
TABLE I
Development of acidity (percentage) in single and mixed cultures of simmered,
and sterilized cow's and buffalo's milk
Duration
(hrs.)
L
C
A
L + C
A + C
A+Y
C+Y
A+Y
+C
Simmered Cow's Milk
12
18
24
0.29
0.38
0.47
0.14
0.20
0.21
0.40
0.77
0.77
0.27
0.37
0.47
0.30
0.38
0.45
0.44
0.63
0.68
0.14
0.19
0.20
0.29
0.41
0.48
Simmered Buffalo's Milk
12
18
24
0.32
0.50
0.50
0.18
0.22
0.27
0.33
0.71
0.91
0.32
0.46
0.50
0.38
0.51
0.51
0.38
0.65
0.84
0.17
0.21
0.25
0.38
0.53
0.54
Sterilized Cow's Milk
12
18
24
0.37
0.38
0.40
0.18
0.23
0.30
0.51
0.54
0.72
0.37
0.43
0.43
0.40
0.50
0.49
0.52
0.57
0.71
0.18
0.18
0.29
0.36
0.39
0.52
Sterilized Buffalo's Milk
12
18
24
0.36
0.38
0.38
0.24
0.25
0.30
0.34
. .
0.61
0.38
0.40
0.44
0.40
0.38
0.47
0.39
0.46
0.54
0.20
0.23
0.29
0.39
0.37
0.63
TABLE II
Periods required for curdling of milk by single and mixed culture starters a
37°C.
Hours
8
. .
. .
. .
. .
. .
12
L . . .
L+C . .
A+Y+L .
C+A+L .
L+C+A+Y .
16
A+C .
A+Y+C
.
. .
. .
. .
20
C .
C+Y.
A+Y.
. .
. .
24
. .
. .
. .
. .
. .
36
A
218
Starters in the Preparation of Dahi
TABLE III
Volatile acidity (as ml. of 0.1 N, NaOH) per 10 ml. of culture in single and
mixed cultures of simmered and sterilized samples of cow's and
buffalo's milk.
—
L
C
A
LC
AC
AY
CY
AYC
Simmered Milk
Cow's . .
Buffalo's .
0.55
0.9
0.7
2.35
1.35
1.80
0.65
1.35
0.7
1.1
Trace
„
0.75
2.25
0.45
0.55
Sterilized Milk
Cow's . .
Buffalo's . .
0.65
0.30
1.05
1.2
0.85
1.6
0.75
1.15
0.75
0.15
„
„
0.85
0.80
0.4
0.05
PLATE XVIII ]
[ Indian J. Vet. Sci. & Anim. Husb., Vol. XII, Part III
FIG.
1. Plant Lupinus angustifolius
FIG.
2. A close view of the plant showing the pods
STUDIES ON LUPINUS ANGUSTIFOLIUS L.
I. A PRELIMINARY INVESTIGATION ON THE USE OF THE SEEDS OF
LUP1NUS ANGUSTIFOLIUS AS CATTLE FOOD
BY
M. R. SUBRAMANIA SASTRY
C. K. VELAYUDHAN NAYAR
R. RAMASWAMY
AND
K. VENKATACHALAM
The Research Unit, Medical College, Madras
(Received for publication on 5 November 1941)
(With Plate XVIII)
L UPINUS ANGUSTIFOLIUS of the natural order Papilionaceae is a
deep-rooted, hardy and spreading bushy annual (Plate XVIII, fig. 1).
It is of a typical papilionaceous variety with the characteristic bacterial nodules
and is a good, green manure plant. Its deep root-system makes it an effective
soil binder. The plant grows to a height of about three feet and has a spread-
ing habit. It gives out a large number of branches with profuse foliage on all
sides. When grown for green manure purpose, the crop yields about 4000 to
5000 pounds of green stuff per acre. The plants show a preference to deep
clayey soils of a low pH value. Flowers are small, axillary, arranged in
racemose fashion, and show all the characters typical of the order. Pods are
thick and stout and about 2 to 2½ in. long with four to five seeds inside (Plate
XVIII, fig. 2). The seeds are round and mottled and have a hard seed-coat.
The yield of seeds varies from 500 to 600 lb. per acre. The seeds are reported
to be a good substitute for horse-gram and useful as cattle food.
The seeds, because of their high protein content (29.74 per cent) according
to Wehmer [1929], have attracted much attention as a valuable food material
for animals. They are also said to contain a variable amount of alkaloids.
The Director of Agriculture, Madras, who was growing a crop of the sweet
lupins at the Agricultural Research Station, Nanjanad, on the Nilgiri Hills
of the Madras Presidency, in rotation with potato with a view to enrich the
soil, wished to know whether the amount of alkaloids present in the seeds
would not prove injurious to the health of animals and whether the seeds from
this crop could, with safety, be utilized for feeding animals. Investigations
were, therefore, undertaken to find out whether these seeds could be safely
fed to animals.
Isolation of lupanine
Eight kilogrammes of the lupin seeds received from the Agricultural
Research Station, Nanjanad, were powdered and extracted with 95 per cent
alcohol containing one per cent hydrochloric acid by the reserved percolation
process. The extraction was continued till the alkaloidal reactions were nega-
tive. The first few fractions of the percolate contained mostly fatty matter.
The alcoholic extract was evaporated and the residual solution was treated
with 3 times its volume of water, kept on a boiling water bath for a few
219
220
Studies on Lupinus angustifolius L.
hours, cooled and filtered. The dark yellow filtrate was neutralized with 15
per cent sodium hydroxide solution, evaporated to a small volume under re-
duced pressure and made alkaline with the same alkali. The alkaline solution
was repeatedly extracted with chloroform till a fraction of the extract gave no
reaction for alkaloid. The chloroform was distilled off and the residual syrup
made slightly acidic with hydrochloric acid and kept in a vacuum desiccator
over NaOH. The yield of lupanine hydrochloride obtained by this method
varied from 1.5 to 2 gm.
Assay of lupin seeds
The method followed for the assay of the seeds was that of Thom and
Michaelis.
15 gm. of the powdered lupin seeds are well shaken with a mixture
of 50 c.c. of ether and 50 c.c. of CHC1 3 . The mixture is treated with 5 c.c. of 15
per cent solution of NaOH and 5 c.c. of water and kept for 24 hours with
occasional shaking. 50 c.c. of ether are then added, and after the mixture has
been shaken and allowed to settle, 50 c.c. of the clear solution are poured off
into a separator. This process is repeated once more. To the combined solu-
tions, a further 50 c.c. of ether is added and the liquid washed 3 times with
about 20 c.c. of water to remove excess of alkali. The ether-chloroform solu-
tion is then shaken with a measured volume (about 30 c.c.) of N/100 hydrochlo-
ric acid and then twice with 20 c.c. of water adding the water to the acid-
extract. The mixture is then titrated with N/100 NaOH using iodeosin as
indicator. A control experiment was carried out without using lupin seed
powder.
The percentage alkaloidal content of the sample of lupin seeds assayed by
the authors by this method is 0.1302. A higher content of the alkaloid has,
however, been reported by certain other workers and the disparity may be
due to the variations in the alkaloidal content of the seeds themselves as appear
to be not improbable from the following findings :—
(i) Hagen
.
.
.
.
.
(ii) Brahmz
.
.
.
.
.
(iii) Schmidt . . . . .
(iv) Beckel . . . . .
(v) Agricultural Research Chemist,
Coimbatore . . .
0.21 per cent to 0.25
per cent
0.292
per cent
0.7249 „ „
0.9 per cent to 1.2
per cent
0.86 per cent
quoted by
Wehmer
[1929]
Probably such variations are due to the nature of the soil on which they
are cultivated, the climatic conditions to which the crops are exposed or the
part of the year at which the crops are harvested.
Biological experiments
Tests were made using a 5 per cent solution of the alkaloid and adminis-
tering it intravenously to dogs weighing about 10 to 15 lb. A dose of 25 to 50
mg. was found to cause a definite fall in the blood pressure and a dose of 100
to 150 milligrammes (or 1½ to 2¼ grains) proved lethal. This dose may corres-
pond to about 18 to 27 grains of the alkaloid orally (about 12 times the intra-
venous dose). This amount of alkaloid represents 1.9 to 2.9 lb. of the raw
M. R. SUBRAMANIA SASTRY AND OTHERS
221
seeds. If the amount of the alkaloid contained in about 2 to 3 lb. of the lupin
seeds should prove lethal to a dog weighing about 10 to 1.5 lb., it may then be
computed on this basis that, in order that it should prove lethal to a bullock
weighing about 300 to 350 lb., the ingestion of the alkaloid contained in 60 to
70 lb. of the seeds will be required.
Feeding experiments
A goat weighing about 45 lb. was fed with lupin seeds. During the earlier
stages of the experiment the seeds were debittered according to the method
adopted at the Agricultural Research Station, Nanjanad, by soaking them in
cold water for about 24 hours, then boiling for one hour and washing in running
water for about 1½ hours. Four ounces (115 grammes) of the seeds were treated
in this way and fed, daily for a period of 16 days, the total consumption being
1.84 kilogrammes or roughly about 4 lb. Though the animal had to be hand
fed for the first two or three days, it began to eat with relish afterwards. The
goat manifested no untoward symptoms and its condition remained normal.
This confirms the results of the feeding experiments done at the Agricultural
Research Station, Nanjanad, where six pairs of work-bullocks were reported to
have been fed, with debittered seeds crushed and mixed with straw, for three
months, daily, at 1 lb. per head and no harmful effects were reported
to have been observed. The animals are said to have maintained good condi-
tion.
Apart from confirming the results of the experiments carried out at Nan-
janad, raw powdered seeds—with the husk intact and without being subjected
to any process of debittering—were also fed to see if any signs of toxicity could
be induced when fed in this way. The quantity used and the number of days
on which feeding was done together with the observations noted are tabulated
below :—
Quantity
given
each
day
(ounces)
4
6
12
16
12
4
No. of
days fed
6 6
6
7
2
2
No fceding
with
lupins—4 days
7
9
10
13
Weight of the goat
lb.
About 45 . . .
Ditto
About 46
About 47
Ditto
Ditto
About 44
About 46
About 47
Observations noted dur-
ing the experiment
Had to be hand-fed for a
few days
...
On the 4th day refused the
feed and so was hand-
fed
...
...
...
...
Had to be hand-fed for a
few days
The goat ate the seeds
with relish
Remarks
The powder was moistened
with water before feeding.
When hand-feeding was re¬
resorted to, it was sus-
pended in water and ad-
ministered with a bottle
Reduced the quantity as the
animal showed satiation