CadmiumToxicity in GrowingSwine 1 ,2
R. J. COUSINS,
University of New Jersey, New Brunswick,
A. K. BARBER ANDJ. R. TROUTÂ»
of AnimalSciences,Rutgers University,
New Jersey 08903
period. Growth rate was decreased as a function of Cd level, having ceased in the 1,350
ppm group.Hematocrit values were the most sensitive
weredecreasedin all Cd-fedanimals.Serum
receiving 450 and 1,350 ppm Cd, while serum calcium was not affected by Cd intake.
Bone ash contentwas decreasedas a function
(LAP)activity was depressedin renal cortex from the groups receiving
more, but serum LAP was unaffectedby Cd intake. The kidney, liver, spleen and teeth
containedthe highest concentrationsof Cd. Kidney Cd increased with dietary Cd level
but appeared to reach a near maximal level in the 450 and 1,350 ppm Cd groups. This
renal Cd contentwas directly relatedto the level of cadmium-binding
from kidney cortex by gel filtration chromatography.
Cadmium(Cd)as cadmiumchloride was addedto the basal diet of
swine at levels of 0, 50, 150, 450 and 1,350 ppm for a 6-week comparison
of toxicity and
of Cd intake. Leucineaminopeptidase
150 ppm Cd or
1973.J. Nutr. 103: 964-972,
Cadmium (Cd ) toxicity has been demon
anemia (1), hypertension
testicular necrosis (5, 6). In humans, epi-
demiologicalevidence has implicated cad
mium in the etiology of hypertension
cardiovasculardisease (8), respiratory
ease (9), and the "Itai Itai" disease (10).
These topics have recently been reviewed
in detail by Friberg and co-workers (11).
There is a paucity of available informa
tion on cadmiumtoxicity in larger mono-
gastric animals, particularly
gations utilizing this species have been pri
marily limited to the ascaricidal activity of
the metal (12).
The objectives of the work presented
this report were: a) to characterize in grow
ing swine the syndrome manifested
posure to graded levels of dietary cadmium;
b) to quantitateas a function of dose the
ability of the tissues to retain
and c) to evaluate
cadmiumon the formation
mium-binding protein (CdBP ).
in numerous animal
of toxicity include
the effect of dietary
of renal cad
in the experiment
initial growing period followed by a 6-week
comparison period. The pigs were weaned
at 6 weeks of age and received a standard
libitum, for a 3-week period prior to wean
ing. After weaningthe animals were fed
the basaldiet (table
ment. At the start of the comparison period,
the animals were 55.3 Â± 5.7 (so)
age. The pigs were randomly
one of five dietary
of basal diet (table
150, 450 or 1,350 ppm cadmium as CdCl2
and were housed in five outdoor, concrete-
floored pens. Diet and tap water, neither
included in each of the first four treatments,
but the 1,350 ppm group was composed of
three animals. Body weights and diet con-
1) thatwas used
of the experi
Received for publication
1Paperof the Journal
Â»Supported by U. S. Public
Inc.. Future Leader
sity of New Jersey.
December 6, 1972.
Grant to Rutgers
College of Agriculture
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BIOCHEMICAL EFFECTSOF CADMIUM
vals. A second experiment, where the same
basal diet (but containing
270 and 810 ppm cadmium)
additionalanimals, was conducted
taneously to aid in analyzing one parame
ter, kidney cadmium content.
At the end of the comparison period, the
animals were killed by exsanguination,
â€”20Â° until analyzed.
mineralanalysis, the organs were rinsed
in deionized water, and the liver, right kid
ney, spleen, lung and brain were homoge
nized, while the heart and right longissi-
mus muscle were minced. The skin samples
were taken from the area directly dorsal to
the longissimus muscle, shaved to remove
the hair and then minced. A minimum of
two sections of the liver, kidney, epipheseal
plate and spleen were stained with hema-
toxylin and eosin for histological examina
tion. Teeth (four medial incisors from the
cleaned and rinsed in deionized water. The
right femur was placed in a bath of boil
facilitateremoval of adhering
distal epiphysis was subsequently
and dried for 1 hour at 105Â°.The bone
samples were then extracted for 48 hours
with a 2:1 mixture of chloroform-methanol
in a Soxhlet extractor
mill (No. 20 mesh).*
Samples for mineral analysis were dried
for 18 hours at 105Â°,then ashed for 48
hours at 450Â°in a muffle furnace. The ash
was dissolved in 3 N HC1 and diluted as
ionized water. Samples for calcium deter
mination contained 0.5% lanthanum oxide.
Mineral assays were performed
limit for cadmium was about 0.005 /Â¿g/ml.
The dry ash procedure
an average Cd recovery with internal stan
dards of 96% Â±1.2 (SEM).
Serum inorganic phosphorus
mined by the method of Fiske and Subba-
Row (13). Hematocrits
with heparinized whole blood. Serum urea
nitrogen was measured
Folin and Svedberg(14) and serum pro
tein was determinedby the biuret method
were measuredat 2-week inter
instead 30, 90,
was fed to 19
for 5 minutes to
and groundin a
by the method of
Composition of basal diet*
Meat and bone meal1'
NaCl and trace mineral premixd
â€¢ Found by analysis to contain 0.73% calcium, 0.15% mag
nesium,85 ppm iron, 6 ppm copper, 20 ppm manganese,
ppm zinc and 0 ppm cadmium.
mum. c Crudeprotein, 17%
NaCl, 96-99%;Mn (manganous
(cobaltcarbonate),0.01%;Zn (zinc oxide),
(calcium iodate), 0.007%.
vitamin Bu, 4.4 mg; riboflavin,
1,940 mg; inarm. 4,740 mg; and choline chloride, 5,280 mg.
bCrude protein, 50% mini
minimum. d Composition :
1,060 mg; d-pantothenic
ployed the liberation
tion of renal leucine aminopeptidase
a 5% homogenate
preparedin 0.001 M Tris-HCl buffer, pH
7.4, and centrifuged
hours at 4Â°. LAP activity was measured on
an aliquot of the supernatant
ployed by Goldberg and Rutenberg
LAP activity in the serum was measured
via the same reaction
centration was determined
of Lowry et al. (17).
ated by gelfiltration
Twenty-eightto 30 g of frozen
cortex were homogenized
speed)with an equal weight
sodium phosphate buffer, pH 7.0, contain
overnight at 4Â°,the homogenate
trifuged at 42,000 X g for 2 hours at 4Â°.
The supernatantsolution was decanted and
recentrifuged as above; both pellets were
discarded.Fifteen to 20 ml of the super
natantsolution was applied
90 cm column of Sephadex G-758 equili-
cortex wasof kidney
at 42,000 X g for 2
as originally em
(16). Protein con
by the method
(1 minute in a
of 0.05 M
to a 2.5 by
*Sif-ma Chemical Co., St. Louis. Mo.
' Model 45. Virtls Co., Inc.. Gardiner. N. Y.
8Pharmacia Fine Chemicals, Inc., Piwcataway, N. J.
Perkln-Elmer Corp., Norwalk, Conn.
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R. J. COUSINS, A. K. BARBER AND J. R. TROUT
13. Fiske, C. H., and Y. SubbaRow
Biol. Chem. 66: 375-400.
methods for the determination
nitrogen, urea, uric acid and sugar in unlaked
blood. J. Biol. Chem. 88: 85.
15. Tietz, N. W. 1970
Chemistry. W. B. Saunders,
16. Goldberg, J. A., and A. M. Rutenberg
aminopeptidasein urine and serum of normal
diseases. Cancer 11: 283-291.
17. Lowry, O. H., N. J. Rosebrough,
and R. J. Randall
ment with the Folin phenol reagent.
Chem. 193: 265-269.
18. Steel, R. G.D., and
Graw-Hill Book Co., New York, p. 109.
19. Jacobs, E. E., M. Jacob,
L. B. Bradley1956
Chem. 223: 147-156.
20. Mustafa, M. G., and C. E. Cross
monary alveolar macrophage.
olism of isolated cells and mitochondria
effect of cadmium ion on electron- and energy-
21. Wilson,R. H., and
portance of diet in studies of chronic toxicity.
Arch. Indust. Hyg. 1: 173-177.
of phosphorus. J.
Fundamentals of Clinical
with cancerand other
A. L. Farr
D. R. Sanadi
22. Hill, C. H., G. Matrone,
C. W. Barber
cadmiumwith copper, zinc and iron. J. Nutr.
23. Wilson, R. H., F. DeEds and A. J. Cox
Pharmacol. Exp. Ther. 71: 222-235.
24. Pond, W. G., P. Chapman
1966Influence of dietary
cadmium on certain blood components,
gain and parakeratosis
Sci. 25: 122-127.
25. Emmerson, B. T.
athy. Ann. Int. Med. 73: 854-855.
26. Mogielnicki,R. P., T. A. Waldmann
olism in experimental
Invest. 50: 901-909.
27. Kagi,J. H.R.,and
Biol. Chem. 235: 3460-3465.
28. Pulido,P., J. H. R. Kagi and B. L. Vallee
1966 Isolation and some properties of human
metallothionein. Biochemistry 5: 1768-1777.
29. Shaikh, Z. A., and O. J. Lucis
tion of cadmium-binding
30. Nordberg, G. F., M. Nordberg,
and O. Vesterberg
forms of rabbitmetallothionein
focusing. Biochem. J. 126: 491^98.
W. L. Payne
In vivo interactions
and E. Walker, Jr.
zinc, corn oil and
in young pigs. J. Anim.
of low renalhandling
I. L-Chain metab
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