NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
184
FLUORINE
INTOXICATION—II
[Vol. XXV, Part III,
The basal ration for all the groups consisted of 1/4 lb. each of groundnut cake,
cotton seed, rice bran and dhall husk as concentrates, with paddy straw supplied
ad lib as roughage. The experiment was commenced on 17th October 1940 and
ended with the death of calf No. 5 on 18th August 1941.
A 4 per cent solution of sodium fluoride was utilised for fluorine adminis-
tration. To start with 25 c.c. of this solution equal to 1 gram of sodium fluoride
was administered to the Groups II and III calves daily in the morning, before the
feed, by drenching with a bottle. This dosage was suitably raised or lowered accord-
ing to the reactions and condition of the animal.
Bone meal was administered mixed with the concentrates. To start with
1 oz. of bone meal per head per diem was fed to calves Nos. 1 to 4, and was
increased gradually to three ounces in the course of six months.
The animals were weighed daily before feeding the concentrates. Periodically,
blood was drawn from the jugular vein for analysis. A metabolism balance experi-
ment was conducted for calcium, phosphorus, and fluorine on the six calves for
four days when all the fluroine-fed animals had developed fully the symptoms
of fluorosis.
RESULTS
Gross symptoms of fluorosis
(a) Growth: The growth curve of the six calves are presented in Fig. 1. From
the data obtained, it will be quite clear that retardation of growth was very marked
in fluorine-fed calves (groups II and III) while the controls (Group I) had steadily
kept on increasing on the same diet and ration.
(b) Legs: The joints in the legs became stiff and swollen after two months of
fluorine feeding. The resulting ankylosis caused impairment of gait and the
animals walked with a laboured staggering gait.
(c) Bones: The long bones, particularly the meta-tarsus and meta-carpus, and
the ribs were found to develop exostosis in a marked manner.
(d) Hoofs: Overgrown and twisted hoofs were also a very marked symptom
in the fluorine-fed calves. This became very characteristic after two months of
fluorine feeding.
(e) Tongue: Towards the end of the experiment, calves Nos. 5 and 6 of
Group III developed a swollen necrotic patch on the upper side of the tongue which
disappeared later in calf No. 6, when fluorine feeding was stopped.
(f) Teeth: Teeth were found to be worn unevenly in all the fluorine-fed calves
but in no case did mottling develop either in the deciduous or permanent teeth,
though chalkiness was present in the incisors of calves No. 4 and No. 5 which were
acutely intoxicated with fluorine.
(g) Appetite and general condition of the animals: Anoerxia and cachetic
condition was very patent in all the fluorine-fed calves. The animals looked very
poor in condition. When the intoxication tended to be acute, diarrhoea was
usually present and the animals became very inert, drowsy and extremely reluctant
to rise or change position, the movements becoming highly painful,
SEPTEMBER 1955]
A. MARIAKULANDAI AND P. VENKATARAMAIAH 185
Fig. 1.—Progressive growth in the control group animals and retardation of growth
in the two fluorine-fed groups
186
FLUORINE
INTOXICATION-II
[Vol. XXV, Part III,
(h) Post-mortem examination: Calf No. 4 died in the middle of the experiment,
i.e. on 22nd June 1941 and calf No. 5 succumbed on 18th August 1941, both due to
fluorine feeding. From the examination, it was found that apart from bone exos-
tosis and overgrown hoofs, which could be seen externally, autopsy revealed acute
inflammation of the gastro-intestinal tract, the small intestines being particularly
congested and highly inflamed.
Effect on blood
(a) Blood inorganic phosphorus and calcium: The results of the periodic
examination made in all the six calves are presented graphically in Figs. 2 and 3.
From the data obtained it will be clear that fluorine feeding does not seem to affect
the calcium or inorganic phosphorus of blood.
(b) Blood fluorine: When no variations from normal in the blood mineral
content of fluorosis calves were noted, it was sought to see whether there was any
variation in the fluorine content of blood. From the data obtained in the latter
half of the experimental period presented in Table I, it was clearly seen that blood
fluorine also does not vary from the control due to fluorine feeding.
TABLE I
Periodic estimation of blood fluorine in the experimental animals
(Mg. per 100 c.c. blood)
Serial
No.
1
2
3
4
5
6
7
8
9
10
11
Date of examination
20th February 1941
6th March 1941
20th March 1941
3rd April 1941
17th April 1941
1st May 1941
15th May 1941
29th May 1941
11th June 1941
24th July 1941
12th August 1941
Group I
Control
Calf I
1.79
1.49
1.25
1.07
1.31
1.13
1.13
1.19
1.37
0.95
1.19
Calf II
1.07
1.31
1.13
1.19
0.96
2.27
1.19
1.19
1.33
1.01
1.34
Group II
Fluorine + Bonemeal
Calf III
1.31
1.19
1.19
1.07
1.19
1.49
2.15
1.49
1.49
1.34
1.97
Calf IV
..
2.15
1.37
1.19
1.01
1.19
1.19
1.25
1.37
..
..
Group III
Fluorine
Calf V
1.49
1.19
1.07
1.01
1.07
1.67
1.19
1.31
1.31
1.79
1.79
Calf VI
1.43
1.07
0.96
0.90
0.90
1.79
1.31
1.07
1.37
1.37
2.03
SEPTEMBER 1955] A. MARIAKULANDAI AND P. VENKATARAMAIAH 187
Fluorine metabolism
Storage of fluorine was computed by working out the fluorine balance in two
ways:
(i) Indirect method: By a metabolism experiment on all the six calves
during their life period by noting fluorine retention
indirectly from the amount of excretion.
(ii) Direct method: By a total metabolism, by noting the amount of fluorine
retained directly in the carcasses of calves No. 1, No. 4
and No. 5 from the total fluorine fed throughout
the experimental period.
(i) Indirect method: Actual fluorine intake in the form of feeds and sodium
fluoride were estimated daily for four days. The daily excretion on these days
in the form of dung and urine were collected separately and the fluorine excreted
in these were estimated.
The intake per diem was as follows:
Calf No. 1
Calf No. 2
Calf No. 3
Calf No. 4
Calf No. 5
Calf No. 6
288 mg. of fluorine
254 „
540 „
528 „
491 „
491 „
The bone meal group animals, calves No. 3 and No. 4, were found to ingest the
maximum amount of fluorine due to the fluorine in the basal diet herein called
'native fluorine'. But most of these 'native fluorine' get excreted as was seen
from the excretion values of the controls. And besides, the 'native fluorine'
did not have any visible effect on the controls.
Fluorine balance: The fluorine balance worked out from the intake and excre-
tion during the experimental period of four days is consolidated and presented in
Table II. The individual ingestion, excretion and balance of fluorine for each
animal is presented in Table VIII. The value of calf No. 1 is taken as representative
of the control group for the preparation of Table II as the other control calf No. 2
was sick during the four days of metabolism experiment.
TABLE II
Fluorine retention by the experimental animals
(As assessed by the indirect method of metabolism experiment)
Group
I. Control
II. Bone meal plus flourine
III. Fluorine
Flourine fed per
day (average of
4 days) in mg.
288
534
491
Flourine retained
per day (average
of 4 days) in mg.
69
264
281
Percentage retained daily
Total
per cent
24
49
57
Excess over
control
per cent
..
25
33
188
FLUORINE
INTOXICATION-II
[Vol. XXV, Part III,
From Table II it will be seen that the control group absorbed 24 per cent of the
fluorine ingested. This fluorine was from the 'native' sources like straw, con-
centrates, etc. The fluorine-fed groups II and III retained 49 per cent and 57
per cent of the total fluorine intake, both from the native sources and the sodium
fluoride fed. From the figures obtained, it will be seen that the animals of the bone
meal group retained on an average 8 per cent lesser fluorine than the pure fluorine
group, which is a factor in favour of bone meal feeding.
(ii) Direct method of working out the fluorine balance: In this, the amount of
fluorine retained was directly estimated in the carcasses. This method of working
out the fluorine balance was adopted in the case of calves No. 1, No. 4 and No. 5,
which represented each of the three groups under study.
Calf No. 4 died in the middle of the experiment and had received 151.3 grams
of sodium fluoride while calf No. 5 which survived for a longer period had received
262.1 gm. of sodium fluoride, during the experimental period. Calf No. 1 which
was slaughtered on the death of calf No. 5 was taken to represent the normal fluorine
content of organs, tissues and bones. The individual analysis of every part of the
animal in the case of calves No. 4 and No. 5, which died due to fluorine intoxication
and calf No. 1 which was slaughtered to get the normal fluorine contents, are pre-
sented in Tables III and IV.
TABLE III
Analysis of organs and tissues
Serial
No.
1
2
3
4
5
6
7
8
9
10
11
12
Organs and tissues
Brain
Lungs
Heart
Stomach
Small intestines
Large intestines
Liver
Kidneys
Spleen
Gall bladder
Musculature
Skin with hairs
Fluorine (mgm. per gram ash)
Control
group calf 1
0.0717
0.3489
0.3071
0.4037
0.3095
0.3301
0.1022
0.2649
0.2201
0.3551
0.2182
0.2601
F+Bonemeal
group calf 4
0.3041
0.4776
0.5967
0.8956
0.9672
0.5939
0.4474
0.6714
0.3915
0.7736
0.4237
0.4851
Fluorine
group calf 5
0.2962
0.6306
0.8143
0.6009
0.7843
0.5051
0.5289
0.7526
0.5982
0.6664
0.8116
0.7456
September 1955] A. MARIAKULANDAI AND P. VENKATARAMAIAH 189
TABLE IV
Analysis of bones, hoofs, etc.
Serial
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Bones, etc.
Incisors
Molars
Mandibles
Lumber vertebrae
Ribs
Humerus
Radio-ulna
Metacarpus
Scapula
Femur
Tibia-fibula
Meta-tarsus
Hoofs
Horns
Fluorine (mgm. per gram ash)
Control
group Calf 1
0.514
0.667
2.150
2.020
1.683
1.759
1.834
1.755
1.735
1.692
1.727
1.455
0.924
1.351
F+Bonemeal
group Calf 4
1.240
2.386
6.640
7.845
4.724
5.284
5.866
6.950
4.724
6.138
4.670
6.427
1.771
4.678
Fluorine
group Calf 5
2.716
3.803
10.860
13.490
14.230
11.160
10.500
11.050
10.700
11.730
9.423
9.610
3.615
5.041
It will be seen from the results obtained that bones stored the highest amount
of fluorine and increased fluorine feeding as in calf No. 5 had caused considerable
increase in the fluorine (nearly 10 times) of bones, with slight increase in tissues
and organs. Among the bones, the cancellated bones like the lumbar vertebrae
and ribs seemed to have a high fluorine content. Teeth, hoofs and horns contain
much less fluorine than the bones. The alimentary canal, musculature, kidneys,
gall bladder and the heart have comparatively more fluorine than other organs and
tissues.
From the individual fluorine analysis of every part of the carcass and the total
weight of each part, the total fluorine retained in the carcass of calves No. 1, No. 4
and No. 5 was worked out and presented in Table V,
190
FLUORINE INTOXICATION-II
[Vol. XXV, Part III,
TABLE V
Total fluorine content of the carcasses of experimental animals
Serial
No.
1
2
3
4
5
6
Parts of the carcass
Skull with teeth
Skeleton
Hoofs and horns
Skin with hairs
Musculature
Internal organs
Total fluorine retained by carcass
Fluorine (in mgm.)
Control
group Calf 1
3361.318
9736.000
7.266
49.510
179.600
35.712
13369.406
mgm.
or
13.369
gm.
Bonemeal
group Calf 4
5657.243
16460.000
8.156
38.590
47.432
79.030
22290.451
mgm.
22.290
gm.
Fluorine
group Calf 5
9397.750
34123.000
12.986
36.560
37.254
112.800
43780.350
mgm.
43.780
gm.
The fluorine retained by the control calf No. 1 was entirely from the 'native'
sources.
As all the three calves received the same kind of feed, the fluorine retained
by calf No. 1 per kilogram body weight, can be taken as due to natural sources
and by subtracting this from the fluorine stored per kilogram body weight by calves
No. 4 and No. 5 the value due to sodium fluoride feeding alone was easily computed.
The results thus obtained are presented in Table VI, the last column representing
the percentage fluorine retained from the sodium fluoride fed to Groups II and III
animals.
TABLE VI
Fluorine retention by the experimental animals
(As assessed by direct analysis of the carcass)
Group
I. Control
(Calf No. 1)
II. Bone meal
plus Fluorine
(Calf No. 4)
III. Fluorine
(Calf No. 5)
NaF fed as
fluorine
(in gm.)
..
69
119
Total
fluorine
retained
(in gm.)
13
22
44
Body
weight
in kgm.
(average
of last
7 days)
173
52
54
FLUORINE RETAINED
Per kgm.
body
weight
(in gm.)
0.075
0.431
0.815
Difference over
control
(in gm.
per kgm.
body
weight.
..
0.356
0.740
From
the NaF
fed
(in gm.)
..
18
40
From
the NaF
fed (per-
centage)
..
26
34
September 1955] A. MARIAKULANDAI AND P. VENKATARAMAIAH 191
From Table VI it will be seen that calf No. 5 had retained more fluorine than
calf No. 4 per kilogram body weight. Percentage retained by calves No. 4 and No. 5
from the total 'Na-Fluorine' fed is presented in the last column of the above
Table. Calf No. 4 of the bone meal group retained 26 per cent of the fluorine fed
from sodium fluoride while calf No. 5 retained 34 per cent; the difference of 8 per
cent lesser storage in the former, being due to bonemeal feeding.
The retention values obtained by the direct method of working out the balance
in the carcass agreed very closely with those obtained in the indirect method.
In both the methods of assessing the retention, the bonemeal group was found to
retain 8 per cent lesser fluorine. Within individual metabolism discrepancies,
the agreement was very close in the percentage retention obtained by either method
of working the fluorine balances, though the indirect method was during the life
period, full four months before the same balance was done directly in the carcase of
the animals. These duplicate values obtained for fluorine retention, by entirely
two different methods of approach showed the lower retention of fluorine in bone-
meal-fed animal.
Bonemeal amelioration
From the live weight chart (Fig. 1), it will be seen clearly that bonemeal
did not prevent retardation of growth in the case of group II animals. As the
initial one ounce dose of bonemeal was found to be ineffective the dosage was in-
creased to 2 ounces and finally to 3 ounces per day during the experiment but to
no avail.
The inherent smell of bonemeal which became too repellent even at 3 ounce
dosage in the concentrate mixture was the chief factor, which stood against any
further increase in bonemeal dosage. Anyway to see whether the bonemeal supple-
ment was adequate in supplementing Ca and P, 40 days after the increase to 3 ounces
of bonemeal, a mineral metabolism experiment for calcium and phosphorus was
conducted on the six calves for four days. The intake, excretion and balance of
these two essential elements are summarised in Table VII.
TABLE VII
Consolidated balance table for calcium and phosphorus
Calf No.
Group I
Group II
Group III
1
2
3
4
5
6
Ingested in gm.
CaO
36.69
35.15
31.43
30.69
6.98
6.98
P 2 O 5
30.21
29.72
28.34
28.05
6.99
6.99
Excreted in gm.
CaO
30.38
34.85
29.56
32.47
10.22
6.89
P 2 O 5
22.97
26.00
23.05
27.04
6.36
4.97
Balance
CaO
+6.31
+0.30
+1.87
—1.78
—3.24
+0.09
P 2 O 5
+7.24
+3.72
+5.29
+1.01
+0.63
+2.02
27 AR/55
5
192
FLUORINE
INTOXICATION-II
[Vol. XXV, Part III,
From the data presented in Table VII, it will be seen that CaO and P 2 O 5 were
much more than what was normally required for the Group I and II animals, while
Group III animals received much below the normal standards, though in a state
of positive balance in regard to P 2 O 5 Thus, the experiment clearly showed that
no further increase in bonemeal dosage was necessary for Group II animals, for they
were receiving more than the stipulated normal standards for young growing bovines
for the elements CaO and P 2 O 5 (viz. 20 grams P 2 O 5 and 20 grams CaO per day).
Thus bonemeal, though fed in dosages much higher than required, did not
prevent symptoms like bone exostosis, overgrown hoofs, etc. from developing
in Group II animals due to fluorine feeding.
The only point in favour of bonemeal feeding was the lesser storage of fluorine
in the system as pointed out under 'Fluorine Metabolism'.
DISCUSSION AND CONCLUSIONS
The results obtained in the present experiment are discussed under the following
main headings:
1. Pathological effects of fluorine feeding
2. Physiological effects of fluorine feeding
3. Improvement by bonemeal feeding
1. Pathological effects of fluorine feeding: Du Toit et al., [1932] induced fluorine
intoxication in adult cattle by sodium fluoride feeding and noted inhibition of growth,
poor milk yield, impaired gait, pronounced infant mortality, bone exostosis and
swollen joints. While Taylor [1929], and Reed and Huffman [1930] also obtained
the same symptoms of fluorine intoxication and irregular wearing down of
the teeth, by feeding rock phosphate. Similarly Phillips, Hart and Bohsted [1934]
also induced fluorosis by rock phosphate feeding and noted in addition to the above
pathological effects, degeneration of the parenchymatous organs. Roholm [1937]
by feeding cryolite and sodium fluoride obtained all the symptoms of fluorine in-
toxication in all its transitional forms between the osteomalacic and osteosclerotic
condition. The symptoms of fluorine intoxication observed in this study on South
Indian calves were in close agreement with previous studies. In man, the most
prominent symptom noticed was the mottling of teeth, while in rats, overgrown
incisors and claws were equally pronounced [Roholm, 1937], but in cattle, teeth do
not seem to figure so prominently in the intoxication symptom. Apart from wearing
down irregularly and turning chalky in advanced toxicosis, mottling was not
observed in the present study either in the deciduous or permanent teeth. However,
the most prominent and as striking a symptom as mottled teeth in man, was the
overgrown and twisted condition of the hoofs in growing cattle. Next in prominence,
was the bone exostosis of the long bones in the limbs and ankylosis. Anorexia
and inanition was a common invariable symptom observed both in this study as
also in other studies by previous workers on induced fluoriosis. Work done in adult
animals since 1941 at the Indian Veterinary Research Institute, Izatnagar,
[Majumdar, Ray and Sen, 1946] has largely confirmed these pathological symptoms
of fluorine feeding.
September 1955] A. MARIAKULANDAI AND P. VENKATARAMAIAH 193
Fig. 2. Periodic concentration of calcium in the blood serum of experimental
animals.
194
FLUORINE
INTOXICATION-II
[Vol. XXV, Part III,
Fig. 3. Periodic concentration of inorganic phosphocus in the blood of experimental
calves.
196
FLUORINE
INTOXICATION-II
[Vol. XXV, Part III,
(e) Mineral metabolism: McClure and Mitchell [1931] concluded from experi-
ments on rats and pigs that calcium retention was probably reduced by fluorine
feeding. Similarly Lanz and Smith [1934] found in rats that fluorine rats excreted
much more calcium and phosphorus than the controls and the retention of these
two important minerals were very much reduced due to fluorine feeding. So also
Du Toit et al., [1937] reported decreased retention in rats, of both calcium and
phosphorous but in bovines they noted slight increase in calcium retention and
pronounced decrease in phosphorus retention.
But Rek [1935] found no such uniform effect on rabbits. The mineral meta-
bolism experiment conducted on the six experimental calves showed no difference
from the normal variation either in excretion or balance due to fluorine feeding.
The result of the experiment can be seen in Table VIII.
TABLE VIII
Fluorine metabolism in the experimental animals
(in mg.)
—
Calf
1
Mean
Calf 2
Calf 3
Mean
Calf 4
Mean
Days of
experiment
22-4-41
23-4-41
24-4-41
25-4-41
..
—
22-4-41
23-4-41
24-4-41
25-4-41
..
22-4-41
23-4-41
24-4-41
25-4-41
..
Intake
288.81
284.91
228.81
288.81
287.84
—
541.81
536.20
541.81
541.81
540.41
517.95
528.78
531.98
534.79
528.38
EXCRETION
In urine
17.80
14.06
33.17
25.83
22.70
Sick
21.33
57.86
60.24
52.12
47.88
13.13
32.28
50.55
32.97
32.23
In faeces
69.37
354.70
173.80
185.60
195.88
—
129.80
184.50
384.50
193.20
223.01
135.80
286.40
229.50
305.90
239.40
Total in
excretion
87.17
386.70
206.97
211.43
218.58
—
151.13
242.36
444.74
245.32
270.89
148.93
318.68
280.05
338.87
271.63
Balance
±
+201.64
— 83.85
+81.84
+77.38
+69.26
—
+390.68
+293.84
+97.07
+296.49
+269.52
+369.02
+210.10
+251.93
+195.92
+256.75
September 1955] A. MARIAKULANDAI AND P. VENKATARAMAIAH 197
TABLE VIII—(contd.)
Fluorine metabolism in the experimental animals
(in mg).
—
Calf 5
Mean
Calf 6
Mean
Days of
experiment
22-4-41
23-4-41
24-4-41
25-4-41
..
22-4-41
23-4-41
24-4-41
25-4-41
..
Intake
491.31
491.31
491.31
491.31
491.31
491.31
491.31
491.31
491.31
491.31
EXCRETION
In urine
18.78
75.09
76.00
89.84
64.93
29.31
85.41
76.02
54.11
61.21
In faeces
82.92
107.70
190.90
161.90
135.85
119.70
115.50
255.20
145.60
159.00
Total in
excretion
101.70
182.79
266.90
251.74
200.78
149.01
200.91
331.22
199.71
220.21
Balance
±
+389.61
+308.52
+224.41
+239.57
+290.78
+342.30
+290.40
+ 160.09
+291.60
+271.10
(g) Fluorine metabolism: Fluorine had been noted to be stored mainly in the
bones and teeth by previous workers. In dogs, Brandl and Tappiener [1891]
showed from complete metabolism throughout the experimental period that nearly
half of the fluorine intake was excreted and that the main fluorine storage was in
the osseous system. Similarly, Sonntag [1917] on dogs; Kick, Bethke and Ed-
gington [1933] and Roholm [1937] on pigs, had demonstrated the higher fluorine
storage in bones, the deposition being in proportion to the daily fluorine intake and
duration of the experiment. Lawrenz, Mitchell and Ruth [1939] found that about
96 per cent of fluorine retained was deposited in skeleton and the remaining 4 per
cent was about equally divided between the teeth and soft tissues. In the present
study also it was demonstrated (cf. Tables IV, V and VI) that the bones predomi-
nantly stored fluorine and the increase in fluorine content was in proportion to the
fluorine fed. For it will be seen that the fluorine content of the bones of calf No. 5
was double that of calf No. 4, as the intake of fluorine in the former was nearly
double that of the latter. The organs and tissues, unlike the bones, do not so easily
increase their fluorine content by fluorine ingestion. Brandl and Tappeiner [1891]
found by analysis of the carcase that out of the 402.9 grams of sodium fluoride fed
to the dogs in their experiment, only 72.6 grams were deposited in the system, the
rest being excreted, i.e. there was a net retention of about 18 per cent. Similarly
Du Toit et al., [1932] found a retention of 31.6 to 34 per cent in bovines. In the
present study on calves out of 69 grams of fluorine fed to calf No. 4, 18 grams, i.e.