NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
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THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, IV
The rationing was based on the use of fodders unusually low in inorganic sul-
phate. Thus, the supplement of sulphate brought up the intake to a value cer-
tainly no greater than that attainable by feeding a sulphate rich fodder such as
Dhub hay without supplement.
The animals received a small amount of groundnut cake, half a kilo of fresh
Guinea grass to ensure a supply of green grass vitamins and a special sample of Bel-
lary hay which was fed ad lib and the daily consumption determined. To avoid
complications due to possible mineral deficiency each sheep was also given 2 gms.
of calcium phosphate daily.
The following figures (Table I) for the sulphur and sulphate content of some
fodders show that the material we were able to employ was specially favourable
for the work.
TABLE. I.
Sulphur and sulphate content of some typical fodders. (All forms of sulphur expressed
as gms. sulphur per cent).
Fodder.
Bellary hay . . .
Dhub grass . . .
Groundnut cake . . .
Guinea grass . . .
Jouar hay . . .
Paddy straw . . .
Ragi straw . . .
Rhodes grass . . .
Total S.
0.070
0.632
0.395
0.127
0.108
0.102
0.151
0.137
Inorganic SO.
0.038
0.453
0.037
0.046
0.045
0.042
0.078
0.057
Organic S.
0.032
0.179
0.358
0.081
0.063
0.060
0.073
0.080
It may be mentioned that this work has been made possible largely because
we were able to employ a special method for the determination of sulphur. The
process is published separately [ Warth & Krishnan, 1935 ].
It should be explained that this sulphur enquiry was superimposed on a simple
feeding test with sheep, in which records were made of the food consumption, the
live weight fluctuations and the digestibility of the rations. The results of the
feeding test are published separately [ Warth & Krishnan, 1935 ], but all the data
from it required for computing sulphur balance have been used hero without
further reference to details.
EXPERIMENTAL.
1. Sulphate balance experiments.
The sulphate balance, being somewhat easier to carry out than a full sulphur
balance, was determined at frequent intervals throughout one year of intensive
study and less frequently during the preceding six months. We have collected
SULPHUR AND SULPHATE BALANCE EXPERIMENTS WITH SHEEP
321
a large number of data. Table II gives typical detailed figures indicating the
amounts of sulphate derived from the different ingredients of the ration and show-
ing how the sulphate balance is computed, for individuals and for the group.
The figures show that all the animals give parallel and similar results and
that the group average data are a fair representation of what takes place in each
test
TABLE II.
Sulphate balance experiments.
Example of a typical test.
(Sulphate expressed as gms. sulphur per day)
Sulphate in
Hay . . .
Guinea grass .
Groundnut cake .
Supplement .
Total intake.
Fæces . .
Digested . .
Excreted in urine
Balance . .
With Supplement Sheep No.
1
0.377
0.054
0.020
0.581
1.032
0.112
0.920
0.594
0.326
2
0.269
0.020
0.017
0.581
0.887
0.073
0.814
0.481
0.333
3
0.301
0.050
0.017
0.581
0.949
0.099
0.850
0.560
0.290
Average
of group
A.
0.316
0.041
0.018
0.581
0.956
0.095
0.861
0.545
0.316
Without Supplement Sheep No.
4
0.382
0.032
0.020
. .
0.434
0.078
0.356
0.176
0.180
5
0.284
0.042
0.017
. .
0.343
0.065
0.278
0.146
0.132
6
0.274
0.050
0.017
. .
0.341
0.066
0.275
0.132
0.143
Average
of group
B.
0.313
0.041
0.018
. .
0.373
0.070
0.303
0.151
0.152
It will be observed that the fæces always contain a small amount of inorganic
sulphate. The fæcal sulphate has been determined with the greatest care. Some
detailed figures are given later, in Table VI. A reference to this table shows that
the fæcal sulphate content is influenced to a determinable extent by the sulphate
intake; but a greatly increased intake causes only a small increase in the fæcal
sulphate. The same conclusion was reached in the bullock experiment. The
D
322
THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, IV
results of all our sulphate balance experiments are given in the next summarised
statement (Table III).
TABLE III.
Summary of sulphate balance experiments.
(Sulphate expressed as gms. sulphur per head per day)
Experi-
ment
No.
1 .
2 .
3 .
4 .
5 .
6 .
7 .
8 .
9 .
10 .
11.
12 .
Average
Ration with sulphate supplement.
Group
A
A
A
B
B
B
B
B
A
A
A
A
Intake
0.956
0.907
0.832
0.933
0.950
0.939
0.960
0.963
0.885
0.850
0.837
0.872
0.909
Outgo
0.640
0.684
0.745
0.722
0.752
0.765
0.726
0.752
0.664
0.657
0.683
0.690
0.707
Balance
0.316
0.223
0.087
0.211
0.198
0.174
0.234
0.211
0.221
0.193
0.154
0.182
0.200
Same ration without sulphate
supplement.
Group
B
B
B
A
A
A
A
A
B
B
B
B
Average
Intake
0.373
0.398
0.280
0.312
0.322
0.309
0.309
0.314
0.342
0.295
0.294
0.321
0.322
Outgo
0.221
0.295
0.288
0.287
0.327
0.287
0.274
0.305
0.241
0.249
0.253
0.245
0.273
Balance
0.152
0.103
—0.008
0.025
—0.005
0.022
0.035
0.009
0.101
0.046
0.041
0.076
0.050
In this table each figure is a group average value calculated from
individual data for the group as in Table II.
The data show that there is almost always a loss of ingested sulphate, the
loss being greater when the amount ingested is increased.
The question of the fate of this sulphate is most important and will be con-
sidered later. Here it may be said, we are inclined to believe, though we have
no direct evidence at present, that this loss of inorganic sulphate occurs chiefly
in the rumen, the sulphate being converted to organic form by bacterial action.
It will be noticed that there are a few instances in which instead of a loss a small
increase of sulphate has occurred. Here, the normal metabolic oxidation of orga-
nic sulphur has exceeded the bacterial reaction, mainly, because the intake of sul-
phate was low. It has to be noted, however, that the sulphate balance as
SULPHUR AND SULPHATE BALANCE EXPERIMENTS WITH SHEEP.
323
determined by us, is the net result of two different processes, the metabolic
production of sulphate and the bacterial conversion to organic sulphur.
It should be remarked, that, in the bullock experiments increase of sulphate
was never observed even with very low intake of inorganic sulphate. In this
respect therefore, cattle seem to be slightly more efficient than sheep.
2. The Total Sulphur Balance.
Altogether six determinations were made during the eighteen months of
quantitative feeding. In each test individual determinations covering ten
days were made with each animal. In the first test there were three animals in
each group, in the remaining tests each group consisted of four animals. Indivi-
duals did show distinctive features, but for the present we must be satisfied
in stating that the individual results, did not differ materially from one another
and the group average results represent very satisfactorily the normal course of
events in each test. As already explained, both groups received the same basal
ration and one group received a supplement of sodium sulphate.
In addition to the total sulphur balance the inorganic sulphate balance was
determined simultaneously and hence, by difference, the organic sulphur balance
was found. The details of a typical total sulphur balance experiment are shown
in Table IV.
TABLE IV.
Details of a typical total sulphur balance experiment.
(All forms of sulphur expressed as gms. sulphur per day).
Sheep No
Intake.
Total
sulphur in
food.
Total sulphate .
Organic sulphur .
Outgo.
Total sulphur in
urine and fæces.
Total sulphate .
Organic sulphur .
Balance.
Loss
of
Total
sulphur.
Loss of total sul-
phate.
Gain
of
organic
sulphur.
With Supplement
1
1.378
0.887
0.491
1.299
0.721
0.578
0.079
0.166
0.087
2
1.227
0.828
0.399
1.104
0.622
0.482
0.123
0.206
0.083
3
1.279
0.856
0.423
1.213
0.681
0.532
0.066
0.175
0.109
4
1.435
0.918
0.517
1.371
0.734
0.637
0.064
0.184
0.120
Average
of group
A.
1.330
0.872
0.458
1.247
0.690
0.557
0.083
0.182
0.099
Without Supplement
5
0.827
0.322
0.505
0.757
0.257
0.500
0.070
0.065
–0.005
6
0.800
0.324
0.476
0.710
0.230
0.480
0.090
0.094
0.004
7
0.745
0.297
0.448
0.698
0.220
0.478
0.047
0.077
0.030
8
0.855
0.340
0.515
0.773
0.271
0.502
0.082
0.069
-0.013
Average
of group
B.
0.807
0.321
0.486
0.735
0.245
0.490
0.072
0.076
0.004
324
THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, IV
The results of the full work, condensed to show group average data only, are
given in Table V.
TABLE V
Total sulphur balance. (All forms of sulphur expressed in gms. sulphur per day
per head)
With Supple-
ment.
Without Sup-
plement.
Group.
A. {
B. {
A. {
Average
B. {
A. {
B. {
Average
Experiment
No.
1
2
5
8
10
12
.
1
2
5
8
10
12
.
Total sulphur
Intake
1.427
1.313
1.486
1.413
1.346
1.330
1.386
0.841
0.859
0.816
0.710
0.824
0.807
0.809
Balance
—0.325
—0.169
—0.186
—0.136
—0.175
—0.083
—0.179
—0.188
—0.105
—0.063
—0.003
—0.077
—0.072
—0.085
Sulphate sulphur
Intake
0.956
0.907
0.950
0.963
0.850
0.872
0.916
0.373
0.398
0.322
0.314
0.295
0.321
0.337
Balance
—0.316
—0.223
—0.198
—0.211
—0.193
—0.183
—0.221
—0.152
—0.103
+0.005
—0.009
—0.046
—0.076
—0.064
Organic sulphur
Intake
0.471
0.406
0.536
0.450
0.496
0.458
0.470
0.468
0.461
0.494
0.396
0.529
0.486
0.472
Balance
—0.009
+ 0.054
+ 0.012
+ 0.075
+ 0.018
+ 0.100
+ 0.042
—0.036
—0.002
—0.068
+ 0.006
—0.031
+ 0.004
—0.023
Sulphur content in
Live-
weight
increase
} 0.013
} 0.021
} 0.014
0.016
} 0.047
} 0.003
} 0.021
0.024
Wool
0.045
0.057
0.059
0.054
0.050
0.047
0.066
0.054
NOTE. — signifies that it is unaccounted for. + signifies that it is in excess of intake.
This is the most important table in which the following points require em-
phasis :
1. There is always a loss of total sulphur.
2. There is almost always a loss of inorganic sulphate.
3. When the amount of sulphate ingested is high (due to supplement)
the loss of sulphate exceeds the loss of total sulphur. Therefore
there is a gain of organic sulphur. This is the most striking fact
brought out by the enquiry.
4. When the ingested sulphate is low, there is on the whole more loss of total
sulphur than sulphate, hence in these circumstances there appears
to be a slight net loss of organic sulphur. The two points on which
special emphasis is laid, viz., the gain of organic sulphur and the
loss of total sulphur will be considered next in greater detail.
3. The Apparent Gain of Organic Sulphur.
Regarding the data on this point in the last table, two facts may be noted.
Firstly, the gain as determined by us is a net gain. Some organic sulphur is oxi-
dised to sulphate, some sulphate is converted to organic sulphur. The net result
SULPHUR AND SULPHATE BALANCE EXPERIMENTS WITH SHEEP.
325
of these opposite processes has been measured. The actual activity of organic
sulphur production was therefore greater than the figures show. Secondly, the
gain recorded does not include organic sulphur stored in wool and in the live-
weight increase. The actual gain is therefore greater than the figures given, by
this amount. The sulphur storage in wool and live-weight increase attained in
our experiment is shown separately in Table V. The figures for wool are accurate
determinations based on wool yields and analyses; the figures for live-weight
increase are based on estimates from the carefully studied live-weight fluctuations.
The gain of organic sulphur may be seen in another way from our figures,
for, feeding of inorganic sulphate has brought about an increase of organic sul-
phur both in the faeces and in the urine as the following data in Tables VI and VII
show.
TABLE VI.
Total sulphur and sulphate in fæces.
(All expressed as gms. sulphur in 100 gms. fæces)
Experiment No.
1 .
2 .
3 .
4 .
5 .
6 .
7 .
8 .
9 .
10 .
11 .
12 .
Average
Sulphate
With
supplement
0.025
0.026
0.022
0.023
0.023
0.022
0.022
0.021
0.019
0.019
0.018
0.019
0.022
Without
supplement
0.018
0.019
0.013
0.019
0.016
0.017
0.017
0.017
0.015
0.015
0.014
0.012
0.016
Total sulphur
With
supplement
0.142
0.155
. .
. .
0.145
. .
. .
0.131
. .
0.130
. .
0.135
0.140
Without
supplement
0.125
0.129
. .
. .
0.132
. .
. .
0.130
. .
0.114
. .
0.107
0.123
326 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, IV
TABLE VI—contd.
Hence average
With supplement
....
Without supplement
Total
sulphur
0.140
0.123
Sulphate
0.022
0.016
Organic
sulphur
0.118
0.107
TABLE VII.
Organic sulphur in urine (mgms. sulphur per day)
Experiment No.
1
2
5
8
10
12
With supplement
24
40
45
63
62
78
Without supplement
24
28
10
20
43
45
Having satisfied ourselves that an appreciable gain of organic sulphur occurs,
we have to consider how and where the gain, which is due to the conversion of
inorganic sulphate to organic sulphur, occurs.
We have no direct evidence but we are inclined to believe at present that the
conversion is brought about by bacteria in the rumen.
That the conversion does not take place at a much later stage, in the lower
intestines, seems to be indicated on the following grounds :
(a) Sulphate entering the duodenum is likely to be absorbed rapidly, thus
leaving little for bacteria to act on in the lower intestines.
(b) The final sulphate content of the fæces seems to be related to the blood
sulphate rather than to the extent of bacterial reduction (unpublished
data on blood sulphate obtained by N. C. Das Gupta of this Section).
(c) As noted in Table VII above, ingestion of inorganic sulphate causes in-
crease of organic sulphur in the urine. This indicates that the or-
ganic sulphur produced from sulphate participates in the general
metabolism and is formed, therefore, in the upper part of the ali-
mentary canal.
SULPHUR AND SULPHATE BALANCE EXPERIMENTS WITH SHEEP.
327
4. The Loss of Total Sulphur.
The loss of total sulphur shown in Table V does not take account of sulphur
stored in the wool and the live-weight increase. The loss is not as great as the
figures of Table V suggest. Allowances which have to be made for the stored sul-
phur are shown in Table VIII
TABLE VIII.
Loss of total sulphur. (In gms. per day.)
With supplement . . .
Without supplement. . .
Intake
1.386
0.809
Balance (loss)
0.179
0.085
Accounted for
in wool & live-
weight
increase
0.070
0.078
Unaccounted
(loss)
0.109
0.007
TABLE IX.
Gain of organic sulphur.
With Supple-
ment.
Without supple-
ment.
Sulphate balance
Intake
0.916
0.337
Balance
(loss)
0.221
0.064
Gain of organic sulphur
Recorded
in Table
V
0.042
— 0.023
Contained
in wool
and live-
weight
increase
0.070
0.078
Total.
0.112
0.055
Un-
accounted
(loss)
0.109
0.009
This statement is an average of all the tests done during eighteen months.
Account has been taken of the live-weight changes and the wool produced by each
individual during three periods. One figure seems to be of special interest:
when sulphate supplement is not added to the ration, the total sulphur in the
food can be accounted for with remarkable completeness. Probably some sulphur
is lost but the loss must be small. On the other hand, when supplement is fed
a large unaccounted loss takes place. Table IX will shed more light on these
facts.
328 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [V, IV
5. Total Gain of Organic Sulphur
It was noted, in discussing the results of Table V, that the gain of organic
sulphur there recorded was not the total gain and should be increased by the
amount of sulphur stored in the wool and the live-weight increase. The total
gain of organic sulphur thus obtained is shown in Table IX. The data of this
table have to be considered from several points of view.
In the first place, to complete the consideration of the loss of total sulphur
discussed above, it is evident now that this loss of sulphur falls on the inorganic
sulphate of the ration. The loss of sulphate, which we know occurs, is accounted
for, according to Table IX, partly, by a conversion to organic sulphur, which
is not a true loss of sulphur but only a conversion to another form. The re-
maining loss is not accounted for. This is a true loss which must take place
by the formation and voiding of gaseous sulphur compounds. The table shows
that both the conversion into organic sulphur and the true loss due to the
formation of volatile sulphur compounds are greatly increased by increasing the
sulphate in the ration.
The second point of considerable interest is that the unsupplemented ration,
very low in sulphate content, which in Table V gave an apparent loss of organic
sulphur, now, when account is taken of the sulphur stored in wool and live-weight
increase, shows that even with this low sulphate intake, there was a net con-
version of some sulphate to organic sulphur, that, in fact, the organic sulphur
balance of the animal on this ration was only maintained by conversion of some
sulphate to organic sulphur. Thirdly, it may be noticed that the sulphur stored
in wool and live-weight increase was actually more without a supplement than
with a supplement. There is, however, no great significance in such a com-
parison. The capacities of the animals were not known when they were grouped.
It happened that one group produced more wool and live-weight increase and
by chance this group spent more time on the unsupplemented ration.
With regard to the organic sulphur available for wool production, the figures
are as follows :—
TABLE X.
Organic sulphur available for wool production
(All quantities are expressed as gms. sulphur per day)
Organic sulphur in food
......
Gain of organic sulphur by conversion of sulphate .
Total .
Organic sulphur in wool
Basal
ration
0.470
0.055
0.525
0.054
Supplemented
ration
0.470
0.112
0.582
0.054
SULPHUR AND SULPHATE BALANCE EXPERIMENTS WITH SHEEP
329
With our animals, therefore, a shortage of organic sulphur for wool produc-
tion should not occur. Under the circumstances it would be useless to look for
any marked effect which organic sulphur formed by conversion of sulphate might
produce.
6. Sulphides in Faeces
We have shown in Table VI that a sulphate supplement to a ration produces
an increase of organic sulphur in the faeces ; but by the method of analysis em-
ployed sulphides would be determined as organic sulphur. Hence it became
necessary to determine sulphides in faeces.
Briefly, the method adopted for this determination is as follows :—The
weighed fresh faeces (about 150 gms.) are placed in a flask with attached reflux
condenser and absorbing bottles. The air is displaced by CO 2 and the apparatus
is rapidly evacuated. Alkali is passed into the absorbing bottles and hydrochlo-
ric acid into the faeces flask (oil is advantageous to prevent frothing) which is
then warmed. Pumping is continued till the contents of the flask are nearly
dry.
The H 2 S absorbed in the alkali can be estimated colorimetrically with lead
acetate but the process was found to be unreliable. We prefer to oxidise the
sulphide with alkaline hypoiodite to sulphate and estimate the same gravimetri-
cally. From a large number of determinations carried out in this way the daily
average sulphide excretion in faeces was found to be as follows:—
TABLE XI.
Without supplement feeding . . . .
With supplement
. . . . . .
Sulphide in
faeces as gms.
sulphur per day
0.0025
0.0048
The supplement produces a marked increase, but the quantities are negligible
in both cases. It is possible that much more sulphide is formed and that it
cannot be retained in the faeces. At all events, of the sulphur remaining in the
faeces, with which alone we are concerned here, the amount present as sulphide
is demonstrated to be negligible. Hence the organic sulphur referred to above
exists in some organic form and is not present as inorganic sulphides.
7. Ethereal sulphates in urine
That the demand on sulphur for detoxication of phenols might sometimes
become an appreciable factor in the sulphur economy was suggested in Warth's
paper in which it was shown that all the para-cresol developed in the body is
E
330 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY. [V, IV
completely conjugated with sulphate. Hence, when the food was deficient in
sulphate the urine did not show a trace of free sulphate ; all was in the ethereal
form. Clearly, the sulphate thus utilised is not available for any other purpose.
Other phenolic bodies were found to have a decided tendency to combine with
sulphate, too, but could only do so when excess of sulphate was present. Later
these facts were used for estimating phenols in urine [Warth and Das Gupta,
1932 ]. Similar results have now been obtained by us in our sheep experiments.
The urines from our unsupplemented ration never showed a trace of free sulphate,
all was present in the ethereal form and the amount may be taken to represent the
conjugation of para-cresol only. The urines from the corresponding supple-
mented ration always contained a large amount of free sulphate and an increased
amount of ethereal sulphate, the increase, as indicated above, being due to con-
jugation of other phenols. The results obtained by us with sheep are shown
in Table XI in which each figure is a group average. In each test the two
groups had identical food, while one group received a sulphate supplement
in addition. The figures show that a very large proportion of sulphur in
the food is employed for the detoxication of the phenols produced during
metabolism.
TABLE XII
Urinary ethereal sulphate per 1,000 gms. dry matter of food consumed, as gms.
sulphur
Experiment No.
1 . . . . . . . . .
2 . . . . . . . . .
3 . . . . . . . . .
4 . . . . . . . . .
5 . . . . . . . . .
6 . . . . . . . . .
7 . . . . . . . . .
8 . . . . . . . . .
Without
Supplement
0.208
0.305
0.338
0.307
0.393
0.345
0.348
0.390
With
Supplement
0.419
0.514
0.567
0.466
0.454
0.460
0.484
0.429
The supplement was fed to group A in the first three tests and to group B
in the remaining ones. The fodders seem to have produced phenols somewhat
more uniformly during the five later tests.
SULPHUR AND SULPHATE BALANCE EXPERIMENTS WITH SHEEP
331
SUMMARY
Sulphur and sulphate balance experiments have been carried out with sheep.
It was found that in the process of alimentation there is always a loss of
total sulphur and sulphate. When the sulphate content of the ration is low the
loss of total sulphur is very small. It was possible in such cases to account for
almost all the ingested sulphur.
On the other hand, when the amount of sulphate in the ration is considerable
a large real loss of sulphur occurs, indicating that volatile sulphur compounds
(H 2 S, etc.), are formed.
In contrast to the losses of total sulphur and sulphate sulphur it was found
that a gain of organic sulphur occurs. This organic sulphur is formed from
sulphate and the amount produced increases with the amount of sulphate
ingested.
That this increment of organic sulphur plays a part in the metabolism
of the animal is indicated by the fact that the organic sulphur content of the
urine is increased by ingestion of inorganic sulphate.
These results confirm and considerably amplify the earlier experiments
with bullocks.
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