Vol. 56 (4) 2001
1
1
2
3
1
1. Department of Biochemistry, Veterinary Faculty, Erciyes University, Kayseri-Turkey
2. Department of Pathology, Veterinary Faculty, Erciyes University, Kayseri-Turkey
3. Toxicology Laboratory, Bornova Veterinary Control and Research Institute, Izmir-Turkey
This study was conducted to determine the changes in biochemical parameters of blood and organs, and tissue mineral levels caused by various levels of dietary cadmium and the effects of zinc on cadmium induced changes in broilers. One hundred forty, day old, broiler chicks were evenly distributed into groups in two experiments with a shared control group, which was fed a basal diet. In experiment I, 50, 75 and 100 mg/kg Cd; in experiment II, 50, 75 and 100 mg/kg Cd plus 40 mg/kg Zn were added to the basal diet. Water and feed were provided ad libitum. In experiment I, live weight was reduced. Damage was observed in the liver, kidney and bursa of Fabricius. Serum total protein and globulin levels, AST, ALT and GGT activities were increased while albumin and phosphorus concentrations were decreased. Dose-dependent correlation was observed between dietary and tissue cadmium levels. In experiment II, zinc did not prevent the adverse effect of cadmium on live weight.
The increased serum total protein and globulin concentrations and ALT and GGT activities were lowered but the activities of these enzymes were still high. Serum calcium levels were decreased and reductions in phosphorus concentrations became more pronounced. Zinc increased tissue zinc levels but had different effects on tissue cadmium. In conclusion, cadmium resulted in suppression of live weight, alterations in biochemical parameters, damage in kidney, liver and bursa of Fabricius, and accumulation in liver, kidney and muscle tissue. The levels of zinc used in this study failed to prevent cadmium toxicity.
Cadmium (Cd) is a heavy metal that is widely distributed in the environment and is present in trace levels in seawater and in a broad range of animal and plant species. Relatively large quantities of Cd are found in commercial phosphate fertiliser, thus the increases in soil and plant Cd contents may lead to increases in dietary Cd. In recent years, Cd poses a potential environmental hazard due to increases in its industrial use (1-3). It was reported that the maximum tolerable dietary Cd level for domestic animals was 0.5 ppm. Dietary concentrations of 1 ppm results in undesirable effects while 5 ppm cause adverse health effects (2).
Gastrointestinal absorption of Cd is affected by the type of diet and nutritional status (3,4).
Absorption of ingested Cd is only about 5% and when absorbed it accumulates first in the liver and then in the kidney (2,5) where its half-life is about 20-30 years (2,3). The high dietary levels of Cd results in suppressed feed intake and weight gain, reduction in bone mineralization and anaemia (2,3).
It was suggested that low levels of Cd induce immunity while high levels suppresses immunity (6). The biochemical alterations occur prior to morphological changes in the organs, and the changes in certain enzyme levels in extracellular fluids may reflect the extent of Cd-induced damage in target organs (5).
In this study, the changes in organs and metabolism, the accumulation of various levels of dietary Cd in tissues, and the effects of zinc (Zn) on Cd-induced changes in broilers were investigated.

Two experiments were conducted with 140 one-day-old Scan-breed broiler chicks. Chicks were evenly distributed in groups in two experiments sharing a control group.
Experiment I
This experiment was conducted to determine the toxic effects and tissue accumulation of various levels of Cd added to the diet. Seventy chicks of both sexes were reared in thermostatically controlled, electrically heated wire separated floor pens on a 24-hour constant light schedule. At one day of age the temperature was set at 35
0
C, then gradually decreased. Chicks were allowed ad libitum access to commercial starter and grower broiler diets ( Table 1 ) and to tap water containing trace levels of Cd and
138 mg/kg, 117 mg/kg and 0.048 mg/L Zn, by analysis, respectively. The diets were supplemented with 50, 75 and 100 mg/kg Cd (CdCl
2
.2H
2
O, Merck). The experiment was started at the same day after hatching and terminated on day 42. At the end of the experimental period, all birds were weighed and blood samples were taken from the wing vein for biochemical analysis. Sera were separated by centrifugation at 3000 rpm for 15 minutes after 1-hour incubation at room temperature. The lactate dehydrogenase (LDH) activities were measured on the same day and sera were stored at - 20
0
C for the remaining analysis. Ten birds from each group were killed by neck dislocation and gross visual examination of organs was made during the necropsy procedure to determine tissue damage and liver, kidney, lung, small intestine bursa of Fabricius, skin, breast muscle and testes samples of 6 birds from each group were collected in 10% neutral-buffered formalin for subsequent histopathological evaluations. In addition, liver, kidney and breast muscle samples of 8 birds from each group were collected and individually frozen in polypropylene bags for mineral analysis.
Serum total protein, albumin, glucose, calcium (Ca) and phosphorus (P) levels, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltranspeptidase (GGT)
(Biocon, Germany), LDH and alkaline phosphatase (ALP) activities were determined spectrophotometrically with commercial kits. Serum Zn (7), feed, water and tissue Cd and Zn concentrations (8) were determined with flame atomic absorption spectrophotometry (Philips Pye
Unicam SP9 Series). Dry ashing procedure was used for the mineral analysis in organs. A 25 g of wet tissue was dried at 100
0
C for 2 hours. Dried samples were transferred to a cool muffle furnace and the temperature was slowly raised to 450
0
C and ashed overnight, then dissolved in 2 ml
HNO
3
plus deionized water. After filtering, the digest was made up to 100 ml with deionized water. Concentrations of Cd and Zn were determined. Serum globulin concentrations were calculated by subtracting the albumin values from total protein values.
The pieces of preserved organs were embedded in paraffin, sectioned (5-6µm), mounted on glass slides, stained with hematoxylin and eosin (H&E) and then examined with a light microscope (9).
Data from both experiments were analysed with SPSS for Windows version 9.0. The differences between groups were determined with one-way ANOVA. When the F values were significant,
Duncan’s multiple range test was performed. The correlation between the doses of Cd added to diet and biochemical parameters were determined. All data were expressed as means ± SEM (10).
Experiment II
This experiment was conducted to study the effect of Zn on Cd-induced alterations in blood chemistry, organs and tissue minerals. Birds received 50, 75 and 100 mg/kg Cd plus 40 mg/kg Zn
(ZnSO
4
.7H
2
O, Merck) added to basal diets from one day to 42 days. Feed and water were supplied ad libitum during the experimental period. The design of the study, the management of the chicks, the samples collected and the analysis procedures performed were the same as described in experiment I.

The effects of Cd and Cd plus Zn on live weight and serum biochemical parameters are presented in Table 2 and tissue mineral levels in Table 3.
No gross lesions were seen on visual examinations of organs during necropsy procedure in both experiments. No pathological changes were observed in any organs of the control group and the lung, intestines, skin, muscle of all birds including the testes in both experiments by light microscopy.
Experiment I
In treatment groups, Cd caused a slight hyperaemia, degeneration, portal fibrosis and increase in Kuppfer cells in the liver
(Fig. 1). Hyperaemia ranged from mild to severe in the vessels of kidney, degeneration in the epithelium of some tubules and cystic dilatation in others with the increased Cd levels. In addition, hyaline casts in the lumen of some tubules and mild fibrosis in interstitial tissue were seen in the groups receiving 75 and 100 mg/kg Cd (Fig. 2). The number of lymphocytes diminished in follicles in bursa of Fabricius. Moreover, disappearance of the cortex-medulla border in groups exposed to 50 and 75 mg/kg Cd, pycnotic alterations in cortical lymphocytes in 50 mg/kg Cd group and lymphocytolysis in 100 mg/kg Cd group were observed (Fig. 3).
Figure 1.
Figure2.

Figure 3.
Experiment II
In the liver, Zn in combination with Cd caused a mild hyperaemia in the 50 and 75 mg/kg Cd and a moderate hyperaemia in 100 mg/kg Cd group. Focal degeneration was observed in hepatocytes in 75 and 100 mg/kg Cd plus Zn groups. In the groups receiving 50 and 100 mg/kg Cd in combination with Zn, fibrosis was seen in portal areas in addition to increases in
Kuppfer cells in 100 mg/kg Cd group (Fig. 4). Hyperaemia ranged from mild to severe in the vessels of kidney was observed in all treatment groups. Degeneration in the tubular epithelium was seen in 50 and 75 mg/kg Cd with Zn groups while this change ranged from degeneration to necrosis in 75 mg/kg Cd group. Cystic dilatation was observed in all groups. In addition, hyaline casts in the lumen of tubules and a mild fibrosis in interstitial tissue were seen in the groups fed Zn with 75 and 100 mg/kg Cd (Fig. 5). In the bursa of Fabricius, the number of lymphocytes in some follicles diminished and hyperplasia was seen in some lymphoid follicles in 50 mg/kg Cd plus Zn group. The number of lymphocytes decreased in the medulla.
Moreover, disappearance in the border of cortex-medulla in groups exposed to 50 and 100 mg/kg Cd plus Zn, necrosis in cortical lymphocytes in 75 and 100 mg/kg Cd plus Zn groups were observed (Fig.6).

Figure 4.
Figure 5.
Figure 6.
Experiment I
In the presented study, live weight of the birds was significantly lower than the controls as in leghorn chicks (3), mice (6) and rats (11) exposed to high dietary Cd. Although feed consumption was not recorded, a reduction was observed in treatment groups as indicated by others (3), the lowered live weight may result from the reduced feed intake.
In the present study, the reductions in serum albumin and increases in globulin levels i.e. alterations in alb/glob ratio may result from the poorer liver function as well as proteinuria due to kidney damage (12) characterised by degeneration in tubular epithelium as well as hyaline casts in the lumen. Since the albumin levels decreased, the increases in globulin levels may elevate the total protein as suggested by the others (12,13). The increases in total protein and globulin levels as indicated by others (13,14,15) may depend on dehydration due to diminished feed and water intake (12). Although, parameters related to immunity were not investigated in this study, in the view of the pathological findings observed in the bursa of
Fabricius, which is the central organ for the production of humoral antibodies in poultry
(16,17), it is possible that Cd had no positive effect on immunity in contrast to the suggestions of Zohouri and Tekeli (13) and Malave and De Ruffino (6), with differences possibly due to the animal species. Cd had no effect on serum glucose concentrations in the present study, which is consistent with the findings of Zohouri and Tekeli (13).

In the present study, increases in AST, ALT and GGT levels (5,11,14,15,18) may result from the liver damage as supported by the pathological findings, characterised by fibrosis in portal area and increased numbers of Kuppfer cells in liver consistent with the findings of
Saygץ et al. (19). A slight but not significant increase in ALP activity in agreement with the finding of Khandelwal et al. (5) may result from poor growth. Although Cd may cause a Zn deficiency due to the antagonism between Cd and Zn, (2), in this study no effects of Cd on serum Zn, a slight but not significant increase in kidney Zn, and significant increases in liver
Zn and the lack of alterations in the activities of serum LDH, which is indicative of Zn deficiency (20) showed that Zn deficiency did not develop.
Cd affects bone development and mineralization (5), and has an antagonistic activity in Ca metabolism (2). However, in the presented study, the serum fluctuating Ca levels could not be explained. With the increasing dietary Cd, the reduction in serum P concentrations may result from either increased P excretion due to tubular damage in kidney (12,21,22), which was supported by pathological findings or suppressed absorption of this element (3) or both. With the increasing dietary Cd, its accumulation increased in all the tissues. The severity of the damage especially in the kidney increased with the increasing dietary Cd indicating that it is more sensitive than the liver to Cd toxicity. Cd accumulated primarily in the liver in consistent with other findings (13,23-25) and also supported the authors indicating that the liver was the primary organ for accumulation of Cd salts while kidney for Cd-metallothionein (Cd-MT)
(26).
Cd concentrations in the tissues of the control group may be a result of the trace levels of Cd in the food and water. Breast muscle Cd levels increased in all groups consistent with the increased dietary Cd levels as indicated by Leach et al. (27).
Experiment II
In the present study, Zn did not reverse the negative effect of Cd on live weight as indicated by Bonner et al. (28). Serum total protein levels diminished in the group fed 100 mg/kg Cd plus Zn. There were no changes in albumin levels, except an increase in the 75 mg/kg Cd plus
Zn group. The globulin values were decreased to control values. Zinc supplementation did affect increased AST activity. The activities of ALT and GGT were still higher than the control. In addition, decreases in Ca and more pronounced reduction P concentrations may result from diminished absorption of these elements and increased excretion due to kidney damage.
The decrease in serum Zn level and a significant increase in kidney Zn observed in 100 mg/kg Cd plus Zn group, and increased kidney Zn levels support elevated Zn excretion found by others (28). It was reported that Cd-treated, zinc supplemented animals had a higher concentration of Cd in the liver (28,29). Similarly, in the present study, liver Cd was higher in the group given 50 mg/kg Cd plus Zn but no effects were observed at higher Cd doses. In contrast to the findings of Bogomazov and Garibian (29), lack of Zn supplementation on kidney Cd levels in this study but is in agreement with the results of Behari and Tandon (30) and Fox et al. (31).
Zinc is one of the nutrients that can reduce the toxicity of orally consumed Cd (2,22,32) and shows its effect by competing with Cd for the same transport system as well as for the binding sites in the MT (6). It was reported that Zn has a protective effect on hepatic cellular damage by maintaining membrane integrity due to its direct action on free radicals (33). Zn given prior to Cd or Cd-MT may prevent the nephrotoxic impact of Cd (33-35) without decreasing kidney
Cd level (34) and the protective effect of Zn was a result of the increased synthesis of MT with the pre-treatment (36). In contrast, no significant effect of Zn on Cd toxicity found in this study may depend on the lack of the time for MT synthesis due to the administration of Zn and
Cd simultaneously.
In conclusion, cadmium resulted in suppression of live weight, alterations in biochemical parameters, damage in kidney, liver and bursa of Fabricius and accumulation in liver, kidney and muscle tissue. When Zn and Cd were given together, the observation of even higher serum enzyme activities, decreases in serum Ca levels and more pronounced decreases in P levels, increases in liver Cd as well as increased severity of kidney damage showed that the level of zinc used in this study failed to prevent cadmium toxicity.

1. Vallee, B.L. and Ulmer, D.D.: Biochemical effects of mercury, cadmium and lead. Annu. Rev.
Biochem. 41: 91-128,1972.
2. McDowell, L.R.: Minerals in Animal and Human Nutrition, Academic Press, New York, pp.
359-361, 1992.
3. WHO: Cadmium. International Programme on Chemical Safety. Environmental Health Criteria
134. Genova, 1992.
4. Yannai, S. and Sachs, K.M.: Absorption and accumulation of cadmium, lead and mercury from foods by rats. Fd. Chem. Toxic. 31(5): 351-55, 1993.
5. Khandelwal, S., Agnihotri, N. and Tandon ,S.K.: Biochemical response to cadmium: dose-time effect. Biol. Trace Elem. Res. 29: 157-164, 1991.
6. Malave, I. and De Ruffino, D.T.: Altered immune response during cadmium administration in mice. Toxicol. Appl. Pharmacol. 74: 46-56; 1984.
7. Anonymous: Introduction to Atomic Absorption Spectrophotometry. Pye Unicom Ltd. Second
Ed. J.W. Ruddock and Sons Ltd., Lincoln, 1983.
8. AOAC: Metals and Other Elements at Trace Levels in Foods, 1998.
9. Luna L.G.: Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology,
3rd Ed. The Blakistan Division, McGraw-Hill Book Company, New York, 1968.
10. D?zg?nes, O.: Istatistik Metotlar ץ, Ankara †niv. Zir. Fak. Yay. No. 578, Ankara †niv.
Bas ץmevi, Ankara, 1975.
11. Rajanna, B., Hobson, M., Reese, J., Sample, E. and Chapatwala, K.D.:Chronic hepatic and renal toxicity by cadmium in rats. Drug. Chem. Toxicol. 7(3): 229-241, 1984.
12. Karag?l, H., Alt ץntas, A., Fidancץ, U.R. and Sel, T.: Klinik Biyokimya. Medisan Yayץnevi
Ankara, 2000.
13. Zohouri, A. ve Tekeli, S.K.: Ratlarda kadmiyumun metabolizma ?zerine etkilerinin arast ץrץlmasץ. Istanbul †niv. Vet. Fak.
14. Chapatwala, K.D., Hobson, M., Desaiah, D. and Rajanna, B.: Effect of cadmium on hepatic and renal gluconeogenic enzymes in female rats. Toxicol. Lett. 12(1): 27-34, 1982.
15. Chapatwala, K.D., Boykin, M., Butts, A. and Rajanna, B.: Effect of intraperitoneally injected cadmium on renal and hepatic gluconeogenic enzymes in rats. Drug. Chem. Toxicol. 5(3):
305-317, 1982.
16. Freeman, B. M.: The Endocrine Status of The Bursa of Fabricius and the Thymus Gland.
Physiology and Biochemistry of the Domestic Fowl. Academic Press, London-New York. Ed.
Bell, D. J. And Freeman, B.M. Vol. 1, 575-587, 1971.
17. Payne, L.N.: The Lymphoid System. Physiology and Biochemistry of The Domestic Fowl.
Academic Press, London-New York. Ed. Bell, D. J. And Freeman, B.M. Vol. 2, 985-1037,
1971.

18. Novelli, E.L., Vieira, E.P., Rodrigues, N.L. and Ribas, B.O.: Risk assessment of cadmium toxicity on hepatic and renal tissue of rats. Environ. Res.79(2): 102-105, 1998.
19. Sayg ץ, S., Deniz, G., Kutsal, O. and Vural, N.: Chronic effects of cadmium on kidney, liver, testis and fertility of male rats. Biol. Trace Elem. Res.31: 209-214. 1991.
20. Keen, C. L. and Graham, T. W.: Zinc. Clinical Biochemistry Of Domestic Animals. Ed. J. J.
Kaneko. Academic Press Inc. New York. Fourth Edition. 1989, pp. 776-784.
21. Leffler, P.E., Jin, T. and Nordberg, G.F.: Nephrotoxic impact of multiple short-interval cadmium-metallothionein injections in the rat. Toxicology. 112: 151-156, 1996.
22. Fox, M.R.S.: Assessment of cadmium, lead and vanadium status of large animals as related to the human food chain. J. Anim Sci. 65: 1744-1752, 1987.
23. Chan, H.M., Zhu, L.F., Zhong, R., Grant, D., Goyer, R.A. and Cherian, M.G.: Nephrotoxicity in rats following liver transplantation from cadmium-exposed rats. Toxicol. Appl. Pharmacol.
123: 89-96, 1993.
24. Patra, R.C., Swarup, D. and Senapi, S.K.: Effects of cadmium on lipid peroxides and superoxide dismutase in hepatic, renal and testicular tissue of rats. Vet. Hum. Toxicol. 41(2):
65-67, 1999.
25. Bose, S., Mukhopadhyay, B., Chaudhury, S. and Bhattacharya, S.: Correlation of metal distribution, reduced glutathione and metallothionein levels in liver and kidney of rat. Indian J.
Expt. Biol. 32: 679-681, 1994.
26. Chan, H.M., Satoh, M., Zalups, R.K.and Chenan, M.G.: Exogenous metallothionein and renal toxicity of cadmium and mercury in rats. Toxicology. 76: 15-26, 1992.
27. Leach, R.M., Wang, K.W. and Baker, D.E.: Cadmium and food chain: the effect of dietary cadmium on tissue composition in chick and laying hens. J. Nutr. 109(3): 437-443, 1979.
28. Bonner, F.W., King, L.J. and Parke, D.V.: The influence of high dietary zinc on tissue disposition and urinary excretion of cadmium, zinc, copper and iron after repeated parenteral administration of cadmium to rats. Toxicology. 19(3): 247-254, 1981.
29. Bogomazov M.I. and Garibian G. M.: The effect of the zinc content in the rations of experimental animals on the absorption, distribution and accumulation of cadmium chloride in the body with different methods of administration. Vopr. Pitan. 4: 51-53, 1992 (Abstract).
30. Behari, J.R. and Tandon, S.K.: Effects of zinc on cadmium, copper and zinc contents in cadmium exposed rats. Toxicol. Lett. 5(2): 151-154, 1980.
31. Fox, M.R., Jacobs, R.M. Jones, A.O. and Fry, B.E.: Effects of nutritional factors of dietary cadmium at levels similar to dose of man. Environ. Health Perspect. 28: 107-114, 1979.
32. Jacobs, R.M., Jones, A.O.L, Fox, M.R.S. and Lener, J.: Effects of dietary zinc, manganese and copper on tissue accumulation of cadmium by Japanese quail. Proceedings of The
Society for Experimental Biology and Medicine.172: 34-38, 1983.
33. Liu, X.Y., Jin, T.Y., Nordberg, G.F., Rannar, S., Sjostrom, M. and Zhou, Y. A.: Multivariate study of protective effects of Zn and Cu against nefrotoxicity induced by cadmium metallothionein in rats. Toxicol. Appl. Pharmacol. 114(2): 239-245, 1992.
34. Dorian, C. and Klaassen, C.D.: Protection by zinc-metallothionein (ZnMT) against cadmium- metallothionein-induced nephrotoxicity. Fund. Appl. Toxicol. 26: 99-106, 1995.

35. Tang, W., Sadovic, S. and Shaikh, Z.A.: Nefrotoxicity of cadmium-metallothionein protection by zinc and role of glutathione. Toxicol. Appl. Pharmacol. 151(2): 276-282, 1998.
36. Scheuhammer, A.M., Onosaka, S., Rodgers, K. and Cherian, M.G.: The interaction of zinc and cadmium in the synthesis of hepatic metallothionein in rats. Toxicology. 36(2-3): 101-
108,1985.
Tables:
Table 1:
Ingredients
Corn
Soy bean meal
Full fat soy
Meat-bone meal
Wheat
Fish meal
Salt
Vitamin-mineral premix*
Total
%
45.742
20.640
20.574
6.852
5.000
0.500
0.350
0.250
100.000
Calculated Nutritional Values %
Crude protein
Digestible crude protein
ME (kcal/kg)
Crude cellulose
Crude fat
Crude ash
Ca
P
Digestible P
Methionine
Sistine
Lysine
Na
Crude protein 21.2
7
Crude cellulose 5.06
Crude ash
Ca
P
Zn (mg/kg)
Cd
6.30
1.00
1.20
138.00
Trace
22.000
18.909
3000.000
3.625
6.634
6.192
1.000
0.802
0.388
0.500
0.405
1.495
0.214
Corn
Soy bean meal
Corn gluten
Meat-bone meal
Fish oil
Fish meal
Salt
Vitamin-mineral premix**
25.825
5.000
4.000
1.607
2.000
0.250
0.250
Crude protein
Digestible crude protein
ME (kcal/kg)
Crude cellulose
Crude fat
Crude ash
Ca
P
18.476
3100.000
3.104
4.759
5.540
1.000
0.600

Methionine
Limestone
Total
0.065
1.016
100.000
Digestible P
Methionine
Sistine
Lysine
Na
0.245
0.400
0.367
1.039
0.149
Crude protein
Crude cellulose
Crude ash
Ca
P
Zn (mg/kg)
Cd
18.07
5.95
6.80
1.04
1.05
117.00
Trace
*: Vitamin-mineral premix provided per kg of diet: Vitamin A, 15 000 IU; vitamin D3, 2000 IU; vitamin E, 20 mg; vitamin K3, 5 mg; vitamin B1, 2.5 mg; vitamin B2, 7.5 mg; vitamin B6, 5 mg; vitamin B12, 0.020 mg, folic acid, 0.75 mg; calcium pantothenate, 10 mg; ascorbic acid, 50 mg; monensin sodium, 100 mg; cholin chloride, 400 mg; nicotinamide, 25 mg; D-biotin 0.05 mg; manganese, 80 mg; iron, 40 mg; zinc, 60 mg; copper, 5 mg; iodine, 0.4 mg; selenium, 0.15 mg; cobalt,
0.1 mg; antioxidant,10 mg.
**: Vitamin-mineral premix provided per kg of diet: Vitamin A, 10 000 IU; vitamin D3, 1200 IU; vitamin E, 20 mg; vitamin K3, 3 mg; vitamin B1, 2 mg; vitamin B2, 6 mg; vitamin B5, 4 mg; vitamin
B12, 0.015 mg, folic acid, 0.35 mg; calcium pantothenate, 5 mg; monensin sodium, 100 mg; cholin chloride, 300 mg; nicotinamide, 25 mg; manganese, 80 mg; iron, 40 mg; zinc, 60 mg; copper, 5 mg; iodine, 0.4 mg; selenium, 0.15 mg; cobalt, 0.1mg; antioxidant,10mg
Table 2.
Experiment I
Parameters Control 50 ppm
Cd
75 ppm
Cd
100 ppm Cd
Experiment II
50 ppm
Cd+ 40 ppm Zn
75 ppm
Cd+ 40 ppm Zn
100 ppm
Cd+ 40 ppm Zn p
Live weight(g (
1467.55
±43.01a
911.60
±
763.30
±
611.25
±
895.55
±
766.15
±
694.20 ±
36.55Cd
39.11b
51.46c
32.75d
47.92b
35.13c
p<0.001
3.49 ±
0.13b
3.38 ±
0.22bc
4.39 ±
0.22a
2.63 ±
0.14d
2.95 ±
0.07Cd
3.00 ±
0.11Cd
p<0.001
Total protein
(g/dl)
3.18 ±
0.10bc
Albumin
(g/dl)
Globulin
(g/dl)
1.34 ±
0.06a
1.90 ±
0.11c
Alb/ Glob 0.77 ±
0.068 b
1.06 ±
0.07c
2.43 ±
0.13b
0.48 ±
0.051bc
0.85 ±
0.05d
2.53 ±
0.21b
0.38 ±
0.037c
1.26 ±
0.09ab
3.13 ±
0.20a
0.39 ±
0.031c
1.14 ±
0.05bc
1.48 ±
0.13c
1.08 ±
0.26 a
1.05 ±
0.05c
1.90 ±
0.08c
0.59 ±
0.047 bc
1.17
±0.05abc
1.83 ±
0.10c
0.69 ±
0.057 bc p<0.001
p<0.001
p<0.001

Glucose
(mg/dl)
AST (U/L)
158.93 ±
6.05
92.25 ±
4.09c
ALT (U/L) 1.60 ±
0.26c
GGT(U/L) 14.50 ±
1.25d
LDH(U/L) 1215.89
± 62.90
127.10
± 16.94
183.31
±
13.75b
4.39 ±
0.90ab
25.97 ±
1.98b
130.25
± 14.57
207.94
±
8.15ab
5.44 ±
1.11a
32.23 ±
1.39a
163.31
± 18.94
222.73
±
10.28a
5.88 ±
0.75a
27.66 ±
1.85ab
156.64
± 14.53
184.51
± 8.99b
4.44 ±
0.95ab
28.25 ±
2.17ab
183.41
± 18.77
190.76
± 9.33b
1.87 ±
0.45c
20.80 ±
1.09c
167.26 ±
21.79
224.64 ±
13.46a
2.73 ±
0.50bc
21.25 ±
1.02c
1160.40
± 33.93
1172.57
± 41.45
1192.00
± 40.30
1292.64
± 53.31
1315.44
± 38.98
1200.51
±
83.81
ALP(U/L)
P (mg/dl) 7.95 ±
0.52a
Ca (mg/dl) 9.31±
0.35bCd
Zn (µg/dl)
1701.23
± 305.03
112.15±
4.65a
1337.52
±
232.04
2002.48
±
377.72
7.17 ±
0.83a
10.66 ±
0.43ab
110.40
± 5.64a
2098.35
±
755.70
4.56 ±
1.47b
7.56 ±
0.64e
114.20
± 3.82a
1957.78
±
398.83
5.69 ±
0.59bc
11.73±
0.39a
110.85
± 6.85a
2570.13
±
320.05
4.99 ±
0.43bc
8.01 ±
0.53de
116.65
± 7.18a
1239.83
± 97.96
4.04 ±
1.42Cd
8.92 ±
0.36Cde
105.15
± 5.15a
3.07 ±
0.28d
9.50 ±
0.51bc
88.55 ±
2.70b
a-e: The mean values within the same row with different superscript differ significantly p<0.001
p<0.001
p<0.001
p<0.001
p<0.001
p<0.01
Table 3.
Muscle
Zn
(µg/g)
Liver Zn
(µg/g)
Kidney
Zn
(µg/g)
Muscle
Cd
(µg/g)
Liver Cd
(µ/g)
Kidney
4.56 ±
0.44c
13.79 ±
0.85c
9.06 ±
0.37c
0.03 ±
0.003f
0.15 ±
0.02f
0.07 ±
Experiment I
Tissue Control 50 ppm
Cd
5.42
±0.24bc
13.02 ±
0.75c
9.26 ±
0.26c
0.09 ±
0.01e
1.82 ±
0.30e
0.91 ±
75 ppm
Cd
5.76 ±
0.31b
0.21 ±
0.02c
2.94 ±
0.19Cd
100 ppm Cd
5.41 ±
0.22bc
22.95 ±
1.3a
11.68±
0.37b
22.61 ±
1.11a
9.70 ±
0.49c
0.39 ±
0.02a
4.43 ±
0.43a
50 ppm
Cd+
40 ppm
Zn
5.67 ±
0.34bc
19.18 ±
1.15b
11.21 ±
0.60b
0.12 ±
0.01de
2.59 ±
0.14d
Experiment II
75 ppm
Cd+
100 ppm
Cd+
40 ppm
Zn 40 ppm
Zn
7.15 ±
0.54a
8.11 ±
0.43a
p p<0.001
20.83 ±
1.06ab
13.58 ±
0.43a
0.17 ±
0.01Cd
3.58 ±
0.17bc
22.21 ±
1.30ab
13.60 ±
0.63a
0.32 ±
0.04b
3.94 ±
0.24ab
p<0.001
p<0.001
p<0.001
p<0.001
1.64 ± 2.14 ± 1.08 ± 1.36 ± 1.89 ± p<0.001

Cd
(µg/g)
0.01f
0.11e
0.09bc
0.24a
0.10de
0.10Cd
a-f: The mean values within the same row with different superscript differ significantly.
0.16ab