NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
September, 1950]
S.
K.
TALAPATRA
187
The data in the above table indicate that the acid-producing property of the
grass had little effect on the consumption of the dry matter. The administration
of wood-ash in the diet to bring about normal alkaline condition had little effect
except that the animals in the wood-ash group consumed on an average about 10 per
cent more of the dry matter.
TABLE IV
Digestibility co-efficients of the whole ration
Animal number
Wood-ash
No wood-ash
1
2
3
4
5
6
Dry
matter
44
47
47
45
45
44
Crude
protein
50
52
52
51
53
51
Ether
extract
60
61
65
57
53
57
Crude
fibre
49
51
51
50
49
48
N-free
extract
46
49
50
47
48
50
Total
carbohyd-
rates
48
50
50
48
50
49
The data shown in the above table indicate that the digestibility co-efficients
of the various organic nutrients are not affected following the feeding of the acid-
producing ulu ; under both sets of feeding, the results are more or less the same. A
careful examination of the data, however, reveals that there is slight but definite
increase in the digestibilities of the ether extract when wood-ash was administered.
The S. E. (Kellner's) and the D. P. have been found to be 10.7 and 0.8 lb. res-
pectively per 100 lb. of the green grass.
TABLE V
Nitrogen balance
Animal number
Wood-ash
No wood-ash
1
2
3
4 5
6
Intake.
gm.
48.73
44.69
47.71
44.42
43.90
42.23
Excretion.
Faeces
gm.
24.26
21.41
22.80
21.57
20.57
20.60
Urine
gm.
15.95
16.92
17.97
18.00
14.20
14.90
Balance
gm.
+8.52
+6.36
+6.94
+4.85
+9.13
+6.73
188
NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XX, Part III
The nitrogen balance is definitely positive and there is hardly any difference
in the faecal and urinary excretion under the two sets of feeding.
The grass ulu at the worst green stage
It has already been shown that the feeding of the grass at the prime stage
along with a concentrate mixture is not likely to do any harm. However, in spite
of all these evidences, it was decided to bring about an extreme condition by feeding
the grass at the worst stage (October and November cut), the concentrate supplement
was also withdrawn. The same animals were taken as experimental subjects. They
were fed exclusively on this grass for about two months. The grass is consumed
at this stage at the rate of 1.6 lb. per 100 lb. body-weight. The digestibility co-
efficients are shown in Table VI. It may be seen from the data that all the animals
showed regularity in digestion and nothing abnormal was noticed.
TABLE VI
Digestibility co-efficients of ulu at the worst stage
Animal
number
1
2
3
Dry
matter
58
62
60
Crude
protein
27
30
32
Ether
extract
32
30
28
Crude
fibre
72
76
73
N-free
extract
54
59
57
Total
carbohyd-
rates
62
67
65
As a result of long continued feeding of late cut ulu, which though induced
acidic urine and brought about unthrifty appearence as is ordinarily observed on
a protein deficient diet but nothing abnormal was noticed. The S. E. (Kellner's)
and D. P. have been found to be 15.6 and 0.44 lb. respectively per 100 lb. of the
raw material.
The crude fibre digestibility of the grass was high when it was fed as a single
feed. It is rather interesting to note that the fibre digestibility was lowered when
fed with a concentrate mixture as shown in Table VII.
The above data will show that in the absence of a miantenance ration, our
animals made a desperate effort to utilise as much of the carbohydrate from the
rass as possible.
The nitrogen balances of the animals under ulu as a single feed are shown in
Table VIII. As expected all the animals showed a high negative balance.
September, 1950]
S.
K.
TALAPATRA
189
TABLE VII
Protein intake and fibre digestibility
Animal number
Wood-ash
No wood-ash
Ulu as a single feed
1
2
3
4
5
6
1
2
3
Protein
per
cent
13
13
3.5
Fibre
digesti-
bility
51
53
53
51
51 50
72
76
73
TABLE VIII
Nitrogen balance under ulu as a single feed
Animal number
1
2
3
In take
gm.
17.5
16.2
14.5
Excretion
Faeces
gm.
12.2
11.4
9.9
Urine
gm.
9.7
11.9
12.5
Balance
gm.
—4.4
—7.1
—7.9
Growing animals as experimental subjects. It is quite clear that the feeding of
the grass to adult animals under a variety of conditions had apparently no bad
effect either on the health or on the metabolism of the organic nutrients. With
a view to furnish further evidences in regard to the suitability of the grass as a
cattle feed, we have carried out a number of experiments with rapidly growing calves,
so that any deleterious effect, however little, which had previously remained either
unnoticed or undeveloped should manifest at least in young animals when fed for a
sufficiently long time. Accordingly three Sindhi calves of about one and a half
year old were taken for the experiments. The early cut ulu was chopped to about
four inches in length and fed to appetite along with a concentrate mixture consisting
of 600 gm. of oil-cake and 600 gm. of rice bran and 100 c.c. of a sodium chloride
190 NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XX, Part III
solution containing 10.8 gm. of Cl 2 . The late or the prime ulu was replaced by the
early cut one, as the young calves did not seem to relish very much the mature
grass. This ration was fed for a period of two months and at the end of which
metabolism experiments were conducted to ascertain the digestibility co-efficients.
The young animals continued to pass urine which was slightly acidic to litmus
right from the fourth day of feeding the grass. No abnormality, however, was
noticed even after feeding for more than two months. When the experiment on
the basic ulu diet was over, the animals were fed 20 gm. of precipitated calcium
carbonate prepared in this laboratory. The feeding of calciun carbonate brought
about some change in the animal system, the urine was alkaline. The animals
were kept on this diet for a period of about one month, at the end of which,
a metabolism experiment was conducted to test the comparative efficiency of the
two sets of feeding. The organic nutriants digested under the two sets of feeding
are shown in Table IX.
TABLE IX
Digestibility co-efficients under acidic and alkaline diet
Animal number
No CaCo 3
CaCo 3
77
89
71
77
89
71
Dry
matter
50
54
56
46
52
51
Crude
protein
45
50
50
50
54
54
Ether
extract
31
31
29
42
40
46
Crude
fibre
56
61
61
56
58
57
N-free
extract
56
58
61
52
51
54
Total
carbohyd-
rates
57
59
61
54
58
56
The data in Table IX tend to show that there is little difference in the digestibility
co-efficients under the two sets of feeding. On an average intake of about 240 gm.
or 0.53 lb. of digestible protein, the animals gained on an average about 51 lb. in
59 days on a basal ration of ulu plus concentrate mixture. This normal growth
apparently indicates that the acidic ulu ration had no detrimental effect on the
general health of the calves. It may be seen from the nitrogen balance data in
Table X that when calcium carbonate was added in the diet of the same animals to
bring about physiologically normal condition, the animals were better balanced but
continued to grow at the same rate. As the period of calcium carbonate feeding
was short, no attempt is made to study the rate of growth under the two sets of
feeding.
September, 1950]
S.
K.
TALAPATRA
191
TABLE X
The nitrogen balance of calves on an acidic and alkaline diet
Animal number
No CaCo 3
CaCo 3
77
89
71
77
89
71
Intake
gm.
76.85
77.22
76.77
75.00
73.62
73.58
Excretion
Faeces
gm.
41.8
38.3
38.6
37.2
34.0
34.0
Urine
gm.
32.7
34.1
31.9
28.8
30.1
24.7
Balance
gm.
+2.35
+4.82
+6.27
+9.0
+9.52
+14.88
II. The metabolism of the inorganic nutrients Ca and P with and without the
Supplementation of calcium salt under various conditions of ulu feeding
It has already been stated that no deleterious effect has been observed as a
result of continuous feeding of the grass either to adult or young animals ; yet,
it is true that the feeding of the grass almost at any stage of growth leads to a sort
of acidosis, the degree of which appears to increase as the plant advances in maturity.
Cattle urine being normally alkaline, attempts have been made to correct the acidic
character of the diet by feeding the cheapest and easily available substances like
wood-ash and calcium carbonate.
We have previously reported that such additions of chemicals in the diet have
no special advantage in so far as the utilisation of the organic nutrients is concerned.
It will be of some interest now to note to what extent the metabolism of the inorganic
nutrients, which are known to be absorbed from the gut under favourable condition
of pH, is influenced by such treatments. Both adult and growing animals were
taken as experimental subjects. The results of these investigations will now be
reported.
Adult animals as experimental subjects. Six local Assamese bullocks were
taken for the experiments. They were fed for a sufficiently long period with ulu
(prime and late cut) balanced by a concentrate mixture, except in one instance
when it was fed as a single feed. The experiments may be described as follows :
The grass ulu (3.5 per cent protein) was fed as a single feed (period 1). It was
then fed at the prime stage (5 per cent protein) with a concentrate mixture with
and without the supplementation of wood-ash (period II). The grass was finally
fed also at the prime stage (5.8 per cent protein) with and without the supplementa-
tion of precipitated calcium carbonate prepared in this laboratory (period III).
The calcium and phosphorus balances as affected by feeding the grass in different
periods are shown in Table XI.
H
192
NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XX, Part III
TABLE XI
Calcium balances as affected by various conditions of ulu feeding
Animal number
Intake
Ulu
gm.
Con-
centrated
mixture
gm.
Added Ca
gm.
Total
gm.
Excretion
Faeces
gm.
Urine
gm.
Total
gm.
Balance
gm.
Period I
Sole feed
1
2
3
8.22
7.61
6.81
..
..
..
..
..
..
8.22
7.61
6.81
7.14
6.11
6.66
1.73
2.24
1.20
8.87
8.35
7.86
—0.65
—0.74
—1.05
Period II
No wood-ash
Wood-ash
1
2
3
4
5
6
10.28
10.07
9.93
12.05
10.39
11.63
1.8
1.8
1.8
1.8
1.8
1.8
..
..
..
5.0
5.0
5.0
12.08
11.87
11.73
18.85
17.19
18.43
14.42
13.38
13.63
21.27
17.89
20.10
1.45
1.30
1.43
1.35
0.84
1.05
15.87
14.68
15.06
22.62
18.73
21.15
—3.79
—2.81
—3.33
—3.77
—1.54
—2.73
Period III
No
CaCo 3
CaCo 3
1
2
3
4
5
6
8.06
8.50
8.47
9.63
9.71
8.13
2.01
2.01
2.01
2.01
2.01
2.01
..
..
..
6.0
6.0
6.0
10.07
10.51
10.48
17.64
17.72
16.14
9.88
11.25
10.58
15.45
15.38
12.88
1.01
1.53
1.90
1.08
1.47
0.47
10.89
12.78
12.48
16.53
16.85
13.35
—0.82
—2.27
—2.00
+ 1.11
+0.87
+2.79
In period I when late cut ulu was fed as a single feed, about 80 and 20 per cent
of calcium could be accounted for in the faeces and urine respectively. This indicates
that about 15 per cent of the absorbed calcium of food origin entered into the blood
circulation, but as the conditions were unfavourable due to low intake of phosphorus
(Table XII) and acidosis, the balances of Ca and P were negative.
In period II when more phosphorus was added in the diet making the Ca/P
ratio 1 : 1, the balances were still negative. This perhaps indicates that the prime
or late cut ulu causes acidosis or creates a condition in the animal body whereby
calcium is absorbed but a positive balance is not assured. The administration of
wood-ash, however, checked acidosis but had no effect on the negative calcium
balance in this period, the faecal calcium was increased by amounts added to the
basal ration, indicating that the mineral from wood-ash was of little consequence.
In period III when precipitated calcium carbonate was added, the alkalinity
and ineffectiveness of wood-ash feeding could be avoided. The animals recorded
definite positive balance, the urine turned slightly alkaline with negligible amounts
of titratable alkali.
September, 1950]
S.
K.TALAPATRA
193
TABLE XII
Phosphorus balances as affected by various conditions of ulu feeding
Animal number
Intake
Ulu
gm.
Con-
centrated
mixture
gm.
Total
gm.
Excretion
Faeces
gm.
Urine
gm.
Total
gm.
Balance
gm.
Period I
Sole feed
1
2
3
4.37
4.04
3.62
..
..
..
4.37
4.04
3.62
5.2
4.6
4.2
0.06
0.06
0.07
5.26
4.66
4.27
—0.89
—0.62
—0.65
Period II
No wood-ash
Wood-ash
1
2
3
4
5
6
5.73
5.61
5.23
6.71
5.79
6.49
6.3
6.3
6.3
6.3
6.3
6.3
12.03
11.91
11.53
13.01
12.09
12.79
13.82
14.33
13.61
16.18
13.60
14.09
0.10
0.07
0.05
0.06
0.15
0.13
13.92
14.40
13.66
16.24
13.75
14.22
—1.89
—2.49
—2.13
—3.23
—1.66
—1.43
Period III
No CaCo 3
CaCo 3
1
2
3
4
5
6
6.06
8.39
6.37
7.24
7.30
6.12
7.64
7.64
7.64
7.64
7.64
7.64
13.7
14.03
14.01
14.88
14.94
13.76
14.00
16.01
15.42
14.01
14.20
12.05
0.07
0.05
0.10
0.10
0.15
0.09
14.07
16.06
15.52
14.11
14.35
12.14
—0.37
—2.03
—1.51
+ 0.77
+0.59
+1.62
The phosphorus balances in Table XII support the figures for calcium shown
in the previous table.
The calcium and phosphorus balances as affected by feeding the grass to growing
animals under various conditions have been shown in Tables XIII and XIV.
H 2
194 NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XX, Part III
TABLE XIII.
Calcium balance of ulu fed calves
Animal number
Intak
Ulu
gm.
Con-
centrated
mixture
gm.
Added Ca
gm.
Total
gm.
Excretion
Faeces
gm.
Urine
gm.
Total
gm.
Balance
gm.
Period I
77
89
71
13.58
13.72
13.55
4.61
4.61
4.61
..
..
..
18.19
18.33
18.16
12.81
11.32
10.61
0.92
1.62
1.60
13.73
12.94
12.21
+4.46
+ 5.39
+ 5.95
Period II
77
89
71
12.94
12.52
12.51
4.42
4.42
4.42
8.0
8.0
8.0
25.36
24.94
24.93
21.09
19.99
19.72
1.07
1.40
1.10
22.16
21.39
20.82
+ 3.20
+ 3.55
+ 4.11
Period III
77
89
71
12.12
13.21
14.00
4.37
4.37
4.37
9.28
9.28
9.28
25.77
26.86
27.65
20.26
19.20
17.93
1.20
1.50
1.20
21.46
20.70
19.13
+ 4.31
+ 6.16
+8.52
In period I when early cut ulu was fed, the animals began to void faintly acid
urine. It may be seen from the data shown in the Table XIII that the animals
retained on an average about 5.3 gm. of calcium in period I. In period II when
calcium carbonate was added on the basal ration, they seem to retain calcium at
the decreased rate of 3.6 gm. per day. The reaction of the urine, however, was
changed from slightly acidic to slightly alkaline condition. The results of the
experiments in period I and II tend to show that an acidic condition of the intestinal
contents is favourable for better retention of calcium in young ones.
However, in order to verify whether increased acidity is tolerated in young
animals, calcium chloride was added in period III, which served two purposes viz.,
(a) it is a highly soluble salt which does not require prior solution before entering
the intestine and (b) it is bound to make the intestinal contents more acidic than
normal, when the basic roughage is either slightly acidic, neutral or slightly alkaline.
The animals were kept on this diet for about a month.
It is quite interesting to note that in period III, the animals retained on an
average as high as 6.3 gm. of calcium per day. The urinary excretion of the mineral
remained practically the same which indicates that up to a certain limit, increased
acidity is better tolerated in young animals than those of the adult ones. In this
connection it may be mentioned that Irving and Ferguson [1925] and Irving [1926]
have shown that an acidic condition of the intestinal contents is more favourable
for the absorption of calcium. In rickets, where considerable disturbances take
place in the absorption mechanism the reaction of the intestinal contents as well as
of the faeces is generally more alkaline than normal [Eastman and Miller, 1935 ;
September, 1950]
S.
K.
TALAPATRA
195
Grayzel and Miller, 1928 ; Oser, 1928 ; Shohl and Bink, 1928]. As is well-known
that the intestinal reaction is restored to neutrality or slight acidity when anti-
rachitogenic agents such as cod-liver oil, Sunlight or ultra violet light is administered.
But under the present conditions with young ulu feeding, the intestinal content is
normally either slightly acidic or neutral and as such seems to be helpful to young
animals for growth and bone-formation. It may be stated that the animals under
the three sets of feeding were in good condition.
TABLE XIV
Phosphorus balance of ulu fed calves
Animal number
Intake
Ulu
gm.
Con-
centrated
mixture
gm.
Total
gm.
Excertion
Faeces
gm.
Urine
gm.
Total
gm.
Balance
gm.
Period I
77
89
71
7.63
7.71
7.61
12.86
12.86
12.86
20.49
20.57
20.47
18.31
17.55
17.17
0.25
0.15
0.20
18.56
17.70
17.37
+1.93
+ 2.87
+ 3.10
Period II
77
89
71
7.63
7.39
7.38
10.12
10.12
10.12
17.75
17.51
17.50
16.57
15.84
15.78
0.15
0.21
0.10
16.72
16.05
15.88
+ 1.03
+ 1.46
+ 1.62
Period III
77
80
71
7.03
7.66
8.12
9.98
9.98
9.98
17.01
17.64
18.10
15.42
14.96
14.76
0.15
0.17
0.20
15.57
15.13
14.96
+ 1.44
+ 2.51
+ 3.14
The phosphorus balances shown in Table XIV support the retention of calcium
almost quantitatively as may be seen from the Table XV :
196 NUTRITIVE VALUE OF THE INDIGENOUS GRASSES OF ASSAM [Vol. XX, Part III
TABLE XV
The calcium and phosphorus balances of calves
Animal number
Balance of calcium
gm.
Balance of phosphorus
gm.
Ca/P
Period I
77
89
71
+4.46
+5.39
+5.95
+1.93
+2.87
+3.10
2.3
1.9
1.9
2.0
Period II
77
89
71
+3.20
+3.55
+4.11
+ 1.03
+ 1.46
+ 1.62
3.1
2.4
2.5
2.7
Period III
77
89
71
+4.31
+ 6.16
+ 8.52
+1.44
+2.51
+3.14
3.0
2.4
2.7
2.7
The data in the Table XV show that in period I when no calcium salt was
added in the ration, the average retention of both calcium and phosphorus was in
accordance with the approximate composition of bone Ca 3 (PO 4 ) 2 . In periods
II and III however, the corresponding retention of calcium was more than that
suggested by the formula Ca 3 (PO 4 ) 2 . On an average, during these periods, about
35 per cent more of the calcium was retained. This increased retention of the
mineral may be of a temporary nature, some of the added salts might have been
mechanically held up somewhere in the rumen.
Further work on the absorption, excretion and utilisation of calcium following
the feeding of acid and alkali-forming cattle feeds is in progress.
SUMMARY
Cattle in Assam thrive almost entirely on grazing. With a view to ascertain
the causes as to why cattle are stunted in growth and poor in production, a survey
of the pasture grasses has been made. It has been found that the feeding of the
predominant species of the pasture grass Imperata arundinacea, Cyrill (ulu) makes
urine acidic at various stages of maturity. It grows abundantly in acid soil and
is poor in protein, calcium, and phosphorus. When frequently cut and new growth
allowed, the stuff becomes softer and better.
September, 1950]
S.
K.
TALAPATRA
197
Metabolism experiments on adult and young animals show that as the grass
advances in maturity, the urine tends to become more and more acidic. When
fed at the early stage, the urine is neutral or slightly acidic and free from bicarbonates.
The administration of wood-ash or precipitated calcium carbonate brings about
normal alkaline condition but has little or no effect on the digestibility co-efficients
and the net utilisation of the organic nutrients of the grass.
The metabolism of calcium and phosphorus is, however, affected by various
conditions of feeding. When adult animals are taken as experimental subjects a
positive retention is not assured with partially mature grass whether it is fed as
a single feed, balanced by a concentrate mixture or supplemented with wood-ash.
When growing animals are fed with the early cut grass and a concentrate mixture,
on an average 5.3 gm. of calcium are retained 20 gm. of precipitated calcium
carbonate on the basal ration makes urine slightly alkaline with an average
retention of 3.6 gm. of the mineral per day. When the calcium carbonate supple-
mentation is withdrawn and replaced by equivalent amounts of calcium chloride
to make the whole ration acidic, an average retention of 6.3 gm. of calcium has
been observed.
All these experiments tend to show that an acidic condition of the intestinal
contents as created by early cut ulu feeding favours rapid absorption of calcium
in young animals. With mature or partially mature grass, adult animals are
likely to suffer from a sort of acidosis.
ACKNOWLEDGMENTS
The acknowledgement is due to the Indian Council of Agricultural Research
for financing the Scheme in Assam. The author is also indebted to R. C. Woodford,
Esq., O.B.E., Director of Agriculture, Assam for his interest and suggestion in the
work. I am also greatly indebted to Shri P. G. Pande, M.Sc., M.R.C.V.S., Principal,
U. P. College of Veterinary Science and Animal Husbandry, Mathura for his helpful
criticism and suggestion.
REFERENCES
Eastman, I. M. and Miller, E. G. Jr. (1935). J. Biol. Chem. 110, 225
Grayzel, D. M. and Miller, E. G. Jr. (1928). J. Biol. Chem. 76, 423
Irving, L. and Ferguson, J. (1925). Proc. Soc. Expt. Biol. Med. 22, 527
Irving, L. (1926). J. Biol. Chem. 68, 513
Iyer, A. V., Ayyar, N. K. and Dasgupta, N. C. (1935). Indian J. vet. Sci. 5, 129
Oser, B. L. (1928). J. Biol. Chem. 80, 487
Sen ,K. C., Roy, S. C. and Talapatra, S. K. (1942). Indian J. vet Sci. 12, 263
Shohl, A. T. and Bink, F. C. (1928). J. Biol. Chem. 79, 269
Talapatra, S. K., Roy, S. C., Kehar, N. D. and Sen, K. C. (1942). Sci. and cult., 8, 209
ASCORBIC ACID CONTENT OF OVARY AND CORPUS
LUTEUM DURING VARIOUS STAGES OF OESTROUS
CYCLE AND EARLY PREGNANCY
By A. ROY, S. N. LUKTUKE and P. BHATTACHARYA, Animal Genetics Section,
Indian Veterinary Research Institute, Izatnagar
(Received for publication on 19 December 1949)
F OR sometime past, information has accumulated to show that ascorbic acid
plays an important role in the regulation of normal reproductive processes.
Follicular and luteal degeneration have been reported in guinea pigs maintained
on scorbutic diet. They also failed to become pregnant, or gave birth to
prematurely born or dead foetus [Kramer et al., 1933]. Observations have been
recorded on the increase in the ascorbic acid content of human fallopian tubes
during pregnancy and the secretory phase of the menstrual cycle. Very little
ascorbic acid was found to be present during the proliferative phase [Joel, 1941].
It has also been reported that ascorbic acid has a progestational effect [Isarael
and Meranze, 1941], as well as a mixed progestational and oestrogenic effect
[Korenchevsky and Hall, 1945] on the endometrium of ovariectomized animals.
Progesterone-like effect of ascorbic acid on the rabbit uterus was, however, not
confirmed by Pratt [1943]. The role of ascorbic acid in regulation of the reproductive
processes of farm animals has evoked much interest since the reports of Phillips
et al. [1940, 1941] that the administration of ascorbic acid to certain impotent or
infertile bulls and sterile cows was followed by the restoration of fertility.
In view of the importance of ascorbic acid on reproductive processes, it seemed
worthwhile to estimate the variation, if any, of ascorbic acid content of ovary and
corpus luteum during various stages of oestrous cycle and pregnancy.
MATERIAL AND METHOD
Both the ovaries of buffalo cows (Bos bubalis) were collected from the abattoir
within half an hour after slaughter and were immediately transported to the
laboratory in jars containing freezing mixture. The ovaries were dissected free
of all extraneous tissues and the corpus luteum was extruded out when present.
The ovarian residue (the residual tissue remaining after the extrusion of its corpus
luteum), the extruded corpus luteum and the other ovary were separately weighed.
The whole of the ovary, ovarian residue and only weighed amount of corpus
luteum were then extracted with a fixed volume of metaphosphoric acid (2 per cent)-
trichloracetic acid (8 per cent) mixture, by grinding the tissue in porcelain mortars
and then filtered. An aliquot of the filtrate was taken for the estimation of ascorbic
acid by the method of Bessy and King [1933]. All estimations were done in tri-
plicate. The results are presented in Tables I to III.
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I