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Animal Feed Science and Technology

139 (2007) 212–233

Influence of dietary inorganic and organic copper

salt and level of soybean oil on plasma lipids,
metabolites and mineral balance of
broiler chickens
M.K. Mondal, T.K. Das, P. Biswas ∗ ,
C.C. Samanta, B. Bairagi
Department of Animal Nutrition, West Bengal University of Animal and Fishery Sciences,
37, Kshudiram Bose Sarani, Kolkata 700037, India

Received 13 July 2006; received in revised form 23 December 2006; accepted 24 January 2007

Abstract

An experiment was conducted to determine the influence of dietary inorganic (copper sulphate) and
organic (copper proteinate) form of copper salt and level of soybean oil on plasma lipids, metabolites
and mineral balance of broiler chickens. Two hundred-day-old commercial Vencobb broiler chicks
were purchased and randomly distributed to 20 cages of 10 birds each. These replicates were randomly
assigned to one of five treatments in a {(2 × 2) + 1} factorial arrangement. Factors were sources of Cu-
salt (Cu-sulphate versus Cu-proteinate) fed at two dietary level (200 mg and 400 mg/kg dietary DM)
and the control (no supplemental Cu). After the starter period (up to 3 weeks), from day 22 onwards
another factor, i.e. levels of soybean oil (0 and 40 g/kg diet) was introduced with the previous factorial
arrangements by subdividing each replicate with two equal parts. Chicks were housed in a battery
type California cages. The birds were given a diet based on maize and soybean meal during starter
and finisher period and the Cu content of the basal diet was 10.5 and 10.8 mg/kg diet, respectively.

Abbreviations: Cu-P, copper proteinate; SBO, soybean oil; HDL-C, high density lipoprotein cholesterol;
LDL-C, low density lipoprotein cholesterol; VLDL-C, very low density lipoprotein cholesterol; ALP, alkaline
phosphatase; ALT, alanine minotransferase; AST, aspertate amino transferase; GGT, ␥-glutamyltransferase; LDH,
lactate dehydrogenase; PCV, packed cell volume; TEC, total erythrocyte count; BW, body weight; CW, carcass
weight
∗ Corresponding author. Tel.: +91 33 25325254; fax: +91 33 25571986.

E-mail address: maicrp@yahoo.com (P. Biswas).

0377-8401/$ – see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.anifeedsci.2007.01.014
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 213

Two birds from each replicate were slaughtered on days 21 and 42 and blood samples were collected
for analyses of plasma lipid and biochemical profiles. Two metabolism trial of 3 days duration were
conducted after days 18 and 39 of feeding trial to determine mineral balance of broiler chicken. Cu-
P supplementation decreased (P<0.01) plasma cholesterol, LDL-C, phospholipids, triglyceride and
blood GSH in comparison to CuSO4 at both days 21 and 42 but increased HDL-C (P<0.01) at day
21. Dose of Cu supplementation showed a linear response (P<0.01) for all the parameters studied
under lipid profiles except total lipids and VLDL-C at both days 21 and 42. Supplementation of
SBO in the diet decreased (P<0.01) cholesterol, HDL-C, triglyceride and VLDL-C; but increased
(P<0.01) phospholipids and blood GSH. Concentration of plasma glucose, proteins, enzymes and
major minerals were similar among the treatments at both starter and finisher period. Concentration
of haemoglobin, packed cell volume, total erythrocyte count and major mineral balance were not
affected by dose and source of Cu-salt at day 21 and by oil, dose and source of Cu-salt at day 42. Dose
of Cu-salt linearly increased (P<0.05) plasma Cu and decreased (P<0.05) plasma Zn level but did not
alter (P>0.10) other plasma trace minerals like Mn and Fe both at days 21 and 42. Supplementation
of Cu-P showed higher (P<0.05) concentration of plasma Cu both at starter and finisher period. SBO
was not found to alter (P>0.10) plasma major and trace mineral concentration at finisher period.
Birds fed diet supplemented with Cu-P linearly increased (P<0.01) retained Cu and decreased Zn
metabolizability compared to inorganic Cu, i.e. CuSO4 . The rate of reduction in retained Zn by Cu
was more in CuSO4 than Cu-P. Effect of addition of SBO reduced (P<0.01) metabolizability and
retained (mg/day; P<0.05) Cu and Zn. Cu-P supplemented birds showed higher (P<0.01) body weight
and carcass weight compared to CuSO4 supplementation. Proportion of abdominal fat was decreased
(P<0.01) with the increased dose of Cu irrespective of Cu-salt at both days 21 and 42 and by the
addition of oil (P<0.05) at day 42. In conclusion, Cu-P supplemented birds showed a reduction of
plasma cholesterol, compared to CuSO4 supplementation. An added level of 400 mg Cu/kg DM was
not toxic to the birds during the feeding trail. Dose and source of Cu had an important role in the
plasma lipids and mineral balance in commercial broiler birds even in presence of soybean oil.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Cu-proteinate; Cu-sulphate; Plasma lipids; Mineral balance; Broiler chicken

1. Introduction

The most commonly used Cu for supplementation in animal diet is inorganic Cu in the
form of Cu-sulphate pentahydrate (CuSO4 ·5H2 O) due to cost and commercial availability.
NRC (1994) requirement of Cu for broiler chickens is only 8 mg/kg. When included at higher
dietary levels, so-called pharmacological levels, this element has found to be effective for
lipid metabolism (Pesti and Bakalli, 1996, 1998; Chowdhury et al., 2004). In recent years,
additional Cu sources have become available and the potential for commercial use as feed
additives has expanded. The bioavailability of Cu inorganic and organic sources has been
shown to differ (Ledoux et al., 1991) suggesting efficacy of them may also differ. Reports
from various workers confirm (Hemken et al., 1993; Du et al., 1996) that metal chelates of
amino acids and peptides can enhance the bioavailability of the trace elements. Manspeaker
et al. (1987) reported that chelated minerals were absorbed more readily than their inorganic
counterpart. Cu is contained in a number of enzymes and proteins, and its role has been
defined for many physiological functions related mostly to a catalytic agent in the active
214 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233

sites of cuproenzymes (McDowell, 1992). Cu has also a great role in lipid metabolism
at pharmacological levels in poultry (Bakilli et al., 1995) and within the physiological
range in finishing steers (Engle et al., 2000). High dietary concentration of Cu increases
liver Cu that regulates cholesterol biosynthesis by reducing hepatic reduced glutathione
(GSH) concentration, which reduces the activity of HMG-CoA reductase, the key enzyme
of cholesterol biosynthesis, thus decreasing cholesterol biosynthesis. Moreover, Konjufca et
al. (1997) indicated that Cu supplementation could increase the metabolism of cholesterol
by cholesterol 7␣-hydroxylase. It is also seen that dietary Cu at pharmacological levels
reduces plasma and breast muscle cholesterol (Bakilli et al., 1995). Numerous papers also
show the absence of effect of source of Cu (Paik et al., 1999; Lee et al., 2001; Paik, 2001)
which made it difficult to know about the extent of alteration of lipid metabolism by dietary
Cu. With this view the present experiment is designed to investigate the influence of level of
dietary inorganic and organic Cu-salt and level of soybean oil on plasma lipids, metabolites
and mineral balance of broiler chickens.

2. Materials and methods

2.1. Experimental birds

Two hundred-day-old Vencobb broiler chicks purchased from a commercial supplier

were used in this experiment. The chicks were individually weighed and allocated to 20
cages of 10 birds (5 male and 5 female) each. These cages were randomly allotted to one
of five treatments of four replicate in a {(2 × 2) + 1} factorial arrangement. Factors were
source of Cu-salt (Cu-sulphate and Cu-P) fed at two dietary levels (200 and 400 mg/kg
dietary DM) and the control, where neither Cu-sulphate nor Cu-P were supplemented.
After starter period (up to 3 weeks), from day 22 onwards day each treatments were again
subdivided into two, resulting 40 replicate with 4 birds each to introduce another factor,
i.e. level of SBO (0 and 40 g/kg diet). Hence the experimental design was converted to
{(2 × 2 × 2) + 2} factorial arrangement. Factors were source of Cu-salt (Cu-sulphate and
Cu-P) fed at two dietary levels (200 and 400 mg/kg dietary DM), level of soybean oil (0 and
40 g/kg diet) and two control (where neither Cu-sulphate nor Cu-P were supplemented),
one with oil and another without oil.

2.2. Supplementation of diet with graded dose of copper

The starter and finisher diets of the experiment were formulated to meet or exceed the
nutrient requirement as per Bureau of Indian Standard (1992) for broiler chicken. Ingre-
dient and chemical composition of all the basal diets were presented in Tables 1 and 2.
Starter feed was given to the birds up to 3 weeks and finisher feed from 4 to 6 weeks.
Upon analysis, the total Cu concentration of the basal diets was 10.57 and 10.86 mg/kg,
respectively. The starter and finisher diet (with or without oil) were supplemented with 200
and 400 mg elemental Cu/kg dietary DM either as CuSO4 (CuSO4 ·5H2 O; Merck Limited,
Mumbai, India; MW: 249.68; Minimum assay 99%) or as Cu-proteinate (as Bioplex Cu
supplied by Alltech Inc., Nicholasville, USA, MW: <1500 Da, minimum assay 15%). To
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 215

Table 1
Composition of basal diets
Ingredients Composition

Broiler startera Broiler finisherb

Without oil With oil

Ingredients, air dry basis (g/kg)
Ground maize 530 625 480
Soya bean meal (440 g CP/kg) 400 315 320
Deoiled rice bran 30 20 119.6
Dicalcium phosphate 18 18 18
Limestone powder 12 12 12
Soybean oil – – 40
Iodized Salt 3 2.9 2.9
NaHCO3 1 1 1
Trace mineral mixturec 1 1 1
Vitamin premixd 1.2 1.3 1.3
Choline chloride 0.5 1.0 1.0
Furazolidone 0.2 0.2 0.2
Maduramycin 0.5 0.5 0.5
Enzymee 0.3 0.3 0.3
l-Lysine (78%) 0.5 0.3 0.3
dl-Methionine (98%) 1.8 1.5 1.5
a Fed from a day old to 3 weeks of age.
b Fed from 3 to 6 weeks of age.
c Provided per kilogram of diet: Zn, 60 mg; Mn, 90 mg; Fe, 110 mg; KI, 2.5 mg.
d Provided per kilogram diet: vitamin A, 10,000 IU; vitamin D , 2500 IU; vitamin E, 25 mg; vitamin K, 1.7 mg;
3
vitamin B1 , 2 IU; vitamin B2 , 5 mg; vitamin B6 , 3 mg; vitamin B12 , 16 ␮g; niacin, 34 mg; calcium-D-pantothenate,
9 mg; folic acid, 1 mg.
e Each kilogram contains: cellulase, 6,000,000 U; xylanase, 10,000,000 U; ␤-glucanase, 700,000 U; ␣-amylase,

700,000 U; protease, 3,000,000 U; pectinase, 70,000 U; phytase, 40,000,000 U.

prepare the graded dose of Cu from CuSO4 , 0.785 and 1.571 g of CuSO4 /kg feed was mixed
with premix (required major and trace minerals mixed with wheat bran) to prepared starter
and finisher diet. Similarly, 1.333 and 2.667 g of Cu-P/kg feed was mixed with premix to
prepare starter and finisher diet. Oil used in the trial was food grade.

2.3. Housing and management

Chicks were housed in a battery type California cages (1.5 in. × 9 in.) with channel feeder
and drinker which were cleaned thoroughly with formaldehyde and potassium permanganate
solution 3 days prior to arrival of birds. The day old chicks were offered electrolyte solution
upon arrival. Birds were maintained on a 24 h constant light schedule. The brooding temper-
ature was maintained close to their requirement, first by heating device for 3 days following
arrival of chicks. Then no additional heating was required as the summer room temperature
was found appropriate up to 3 weeks and finally by turning cooler fan during day time
for the last 3 weeks of rearing period. The birds were vaccinated against Ranikhet disease
(Newcastle) and Infectious Bursal Disease on 7, 14 and 21 days and provided antibiotic
216 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233

Table 2
Chemical composition, calculated
Chemical composition (DM basis)a Broiler starter Broiler finisher

Without oil With oil

ME (MJ/kg)b 11.73 12.13 12.22
DM (g/kg) 894.5 905.4 907.4
CP (g/kg) 217.2 191.8 192.1
EE (g/kg) 28.8 27.4 67.9
CF (g/kg) 41.1 41.6 46.2
Ash (g/kg) 84.2 82.1 84.4
Calcium (g/kg) 9.8 9.6 10.2
Total phosphorus (g/kg) 6.1 6.2 6.5
Mg (g/kg) 19.1 19.5 19.8
Cu (mg/kg) 10.57 10.94 10.79
Zn (mg/kg) 81.72 82.01 81.81
Mn (mg/kg) 110 117 121
Fe (mg/kg) 296 312 316
Methionine (g/kg)b 6.8 6.0 6.1
Methionine + cystine (g/kg)b 8.5 7.6 7.5
Lysine (g/kg)b 12.2 10.1 10.2
Available phosphorus (g/kg)b 5.12 4.93 5.28
a Assayed values.
b Calculated on the basis of standard values applicable under Indian Condition (Singh and Panda, 1996).

for the first 5 days as per recommendation. Total amount of feed offered during 24 h to a
replicate under a specific treatment groups was divided into three equal proportions. The
amount and timing of feed was adjusted in such a way that the birds consume the whole of
the diet offered at any one time. As a result hardly any residue can be obtained from the
replicate after a days feeding. These ensured the consumption of the mineral elements that
might have been precipitated in the dust portion of the feed.

2.4. Metabolism trial and sample analysis

Two metabolism trial of 3 days duration were conducted after days 18 and 39 of feed-
ing trial to determine mineral balance of broiler chicken. During the metabolism trial total
amount of feed consumed and total amount of excreta voided from each replicate of the
individual experimental group was quantified. The excreta samples (100 g) from each repli-
cate was collected everyday in a previous weighed clean and dry petridish and were dried
in hot air oven (100 ± 2 ◦ C), pulled together and preserved for subsequent analysis.
The dietary ingredients were dried at 70 ◦ C for 12 h in a hot air oven and ground to
pass through a 1 mm sieve, were analyzed for DM, ether extract (EE), crude fiber (CF)
and total ash (AOAC, 1995, ID 7.010, 7.016, 7.048). The nitrogen fraction of feed samples
was estimated in an automatic analyzer (Kjeltech Auto 147 1013 Analyzer, Foss Tecator,
Sweden) and crude protein (CP) was determined by multiplying the N value with 6.25. For
analysis of major (Ca, P and Mg) and trace (Cu, Zn, Mn and Fe) mineral concentration
of feeds and excreta, samples were dried in hot air oven at 105 ◦ C for 8 h, ground to
pass through 0.5 mm sieve and transferred to a glazed ceramic crucible. The samples were
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 217

ignited at 400 ◦ C for 4 h in a muffle furnace. The ash was treated with concentrated HNO3
under mild heat. After complete digestion, the acids were cooled at room temperature and
filtered through Whatman grade 1 filter paper (AOAC, 1995, ID 7.073). The crucibles
were washed several times with triple distilled water and final volume was made up to
50 mL. Subsequent analysis of major and trace minerals were estimated by flame Atomic
Absorption Spectrophotometer (A Analyst 100, Perkin-Elmer Inc., USA) after suitable
dilution. P content of feeds and excreta was determined as per AOAC Method 7.076 (AOAC,
1995).

2.5. Collection and processing of blood samples

Two birds (one male and one female) from each replicate were slaughtered on days 21
and 42 by severing the carotid artery and jugular vein and blood samples were collected for
analyses of lipid and biochemical profiles. Blood samples (approx. 10 mL) collected in two
sets of heparinized vacuitainer tube (Becon Dickinson India Pvt. Ltd., New Delhi, India);
one for haematological and another for biochemical study. Immediately after collection,
tubes for haematological study were placed in refrigerator (4 ◦ C) and analyzed subsequently.
Other sets were placed in an ice bath and transported to the laboratory. Plasma was harvested
subsequently by centrifuging the whole blood samples at 3000 rpm for 15 min in centrifuge
machine. The heparinized plasma samples were stored at −20 ◦ C in Eppendorf tubes and
analyzed subsequently.

2.6. Chemical analysis of blood samples

Heparinized blood samples were used for the determination of haemoglobin, packed cell
volume (PCV) and total erythrocyte count (TEC). Haemoglobin was estimated from the
whole heparinized blood samples by cyanmethaemoglobin method (Cannan, 1958). PCV
of blood samples were determined in Wintrobe haematocrit tube as per standard method
of Schalm et al. (1975). TEC was enumerated by haemocytometer as per standard method
of Schalm et al. (1975). Plasma samples were analyzed for lipids {plasma total choles-
terol, high density lipoprotein cholesterol (HDL-C), low density lipoproteins cholesterol
(LDL-C), very low density lipoprotein cholesterol (VLDL-C) total lipids, triglyceride and
phospholipids}, glucose, proteins (total proteins, albumin and globulin), enzymes {alkaline
phosphatase (ALP), alanine aminotransferase (ALT), aspertate aminotransferase (AST), ␥-
glutamyltransferase (GGT), lactate dehydrogenase (LDH) and reduced glutathione (GSH)}
and minerals {major (Ca, P and Mg) and trace (Cu, Zn, Mn and Fe) minerals}. Plasma
total cholesterol (Chod Pod enzymatic method, ref. 30180), HDL-C (phosphotungstic acid
method, ref. rekt 106), LDL-C, VLDL-C (colorimetric method, ref. 30185), triglyceride
(Gpo Pod enzymatic colorimetric method, ref. 30360) phospholipids (trinder method, ref.
30210) and total lipids (sulpho-phosphovailline method, ref. 30345) were analyzed in the
Automatic Blood Analyzer (Microlab 200, E-Merck India Ltd., Mumbai, India) using com-
mercial kit (Labkit, Chemelex, S.A., Spain). Plasma glucose (God Pod method, code. rekt
201–203), total protein (Biuret method, code. rekt 391), albumin (BCG dye method, code.
rekt 061), AKP, ALT, AST, LDH and GGT (IFCC method, code FBCEM0020/19, 28 and
29) were analyzed in the Automatic Blood Analyzer (Microlab 200, E-Merck India Ltd.,
218 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233

Mumbai, India) using commercial kit (Transasia Bio-Medical Ltd., Ringanwada, Daman,
India). Blood reduced glutathione (GSH) were measured by the method of Beutler et al.
(1963), which is based on the development of a relatively stable yellow colour when DTNB
[5,5 -dithiobis-(2-nitrobenzoic acid)] is added to sulphhydril compounds. Plasma minerals
except P were determined as per the method described by Sandel (1950) and modified by
Arenza et al. (1977) utilizing atomic absorption spectrophotometer (Perkin Elmer A Analyst
100). Plasma P was estimated by the method described by Fiske and Subbarow (1925).

2.7. Carcass dissection and abdominal fat collection

Two birds from each replicate were slaughtered on days 21 and 42 and dissected into
different parts for cut-up part studies. After removal of viscera, the abdominal fat pad (from
the proventriculus surrounding the gizzard down to the cloaca) was removed and weighed
(Cahaner et al., 1986) which is indicative of abdominal fat content.

2.8. Statistical analysis

The experiment was divided into two parts, i.e. starter and finisher. There were two
factors up to 3 weeks, i.e. salt and dose of Cu but-salt another factor, i.e. supplemental SBO
was introduced after 3 weeks. So up to 3 weeks data were collected and were analyzed
by general linear model of SPSS (1997) with replicates as experiment units. Data obtained
during this period were analyzed separately to determine the main effects of Cu-salt and
dose of supplemental Cu at day 21. Salt and dose interaction was also determined. For this,
Cu-salt and dose were used as fixed factor in this model. Polynomial contrast (linear and
quadratic) was applied to determine the effects for different dose levels (0 mg/kg versus
200 and 400 mg/kg) of supplemental Cu. Data of finisher phase were analysis by the same
model to determine the main effect of SBO, Cu-salt and dose of supplemental Cu, where
salt, dose and SBO was taken as fixed factor. A probability of P<0.05 was considered to be
statistically significant.

3. Results

3.1. Effect on plasma lipids

3.1.1. Starter period

Concentrations of plasma lipid profiles, blood GSH and body weight of broiler chickens
supplemented with two sources of Cu-salt at day 21 are presented in Table 3. Cu-P supple-
mentation decreased (P<0.001) plasma cholesterol, LDL-C, phospholipids, triglyceride and
blood GSH in comparison to CuSO4 and control but increased HDL-C (P<0.001). The effect
of Cu-P on total lipid and VLDL-C was comparable (P>0.10) with CuSO4 supplementation.
Dose of Cu-salt supplementation showed a linear response (P<0.01) for all the parameters
studied under lipid profiles except total lipids and VLDL-C. Significant quadratic effect of
dose of Cu-salt (P<0.05) was found for HDL-C and LDL-C. Four hundred milligrams Cu
per kilogram feed irrespective of source decreased (P<0.01) cholesterol, triglyceride and
LDL-C concentration than 200 mg Cu/kg dietary level.
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233
Table 3
Concentrations of plasma lipid profiles, blood reduced glutathione and body weight in broiler chicken supplemented with two sources of Cu-salt (Cu-sulphate and
Cu-proteinate) at day 21 of Cu supplementation (each group having four replicates)
Attributes Control, 0 Source and dose of supplemental S.E. Significance of treatment
copper mg/kg diet DM effects (P > F)
Copper sulphate Copper proteinate Source Dose
200 400 200 400 Linear Quadratic
Body weight (g/bird) 552 576 584 581 596 1.9 <0.001 <0.001 0.005
Cholesterol (mmol/L) 3.56 3.25 3.06 3.15 2.87 0.028 <0.001 <0.001 0.150
HDL-cholesterol (mmol/L) 1.87 1.76 2.05 1.96 2.32 0.019 <0.001 <0.001 0.049
LDL-cholesterol (mmol/L) 0.89 0.74 0.62 0.70 0.58 0.010 0.001 <0.001 0.042
LDL-C/HDL-C 0.39 0.36 0.33 0.36 0.33 0.003 0.353 <0.001 0.869
Triglyceride (mmol/L) 0.84 0.77 0.70 0.73 0.66 0.009 0.002 <0.001 0.499
Total lipid (mg/dL) 344 345 340 342 338 1.2 0.312 0.358 0.282
Phospholipids (mmol/L) 131 128 127 125 123 0.7 <0.001 0.001 0.152
VLDL-cholesterol (mmol/L) 0.09 0.09 0.09 0.09 0.09 0.005 0.850 0.888 0.783
Blood GSH (mg/100 mL) 69.44 66.56 62.41 64.23 60.11 0.376 <0.001 <0.001 0.941

219
220 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233

3.1.2. Finisher period

Concentrations of plasma lipids, blood GSH and body weight of broiler chicken
supplemented with SBO and two sources of Cu-salt at day 42 of Cu supplementa-
tion are presented in Table 4. Supplementation of SBO in the diet decreased (P<0.01)
cholesterol, HDL-C, triglyceride and VLDL-C; but increased (P<0.01) phospholipids
and blood GSH. Supplementation of Cu-salt had no effect (P>0.10) on plasma HDL-
C, triglyceride and VLDL-C concentrations. Cu-P salt supplementation showed better
response in respect to reduction in plasma cholesterol, LDL-C and phospholipids concen-
tration. Linear response (P<0.01) of dose of Cu-salt was found for plasma cholesterol,
HDL-C, LDL-C, LDL/HDL, phospholipids and blood GSH. Quadratic effect of dose
of Cu-salt was found (P<0.01) only for plasma cholesterol and HDL-C concentra-
tion.

3.2. Effect on plasma glucose, protein and enzymes

The concentration of plasma glucose, proteins and enzymes of broiler chicken was unaf-
fected by source and dose of Cu-salt at day 21 and oil and source dose of Cu-salt at day
42. The overall treatment means at days 21 and 42 were 9.20 and 11.41 mmol/L (glucose),
37.13 and 39.76 g/L (total protein), 22.17 and 23.91 g/L (albumin), 14.42 and 15.85 g/L
(globulin), 5.95 and 6.05 ␮kat/L (alkaline phosphatase), 1.82 and 1.83 ␮kat/L (aspertate
aminotransferase), 0.76 and 0.78 ␮kat/L (alanine aminotransferase), 7.50 and 6.57 U/L (␥-
glutamine transferase) and 6.67 and 6.35 ␮kat/L (lactate dehydrogenase). Concentration
of these plasma enzymes signified that the birds were apparently healthy throughout the
experimental period.

3.3. Effect on haematological profile

Whole blood haemoglobin (g/dL), packed cell volume (PCV) and total erythrocyte count
(×106 mm−3 , TEC) throughout the starter and finisher period were similar among the
treatments and their mean concentration were 10.67, 30.10, 3.97 and 10.81, 30.15, 4.13 at
days 21 and 42, respectively.

3.4. Effect on plasma minerals

Plasma major mineral (Ca, P and Mg) concentration were unaffected (P>0.10) by dietary
treatments both at days 21 and 42. On days 21 and 42 of the study, average plasma major
mineral concentrations were 2.15, 1.40, 3.58 and 2.08, 1.41, 3.75 for Ca (mmol/L), P
(mmol/L) and Mg (mg/dL), respectively. Concentration of trace minerals of broiler chicken
at starter and finisher period under different treatments were presented in Table 5. Effect
of source and dose of Cu-salt on plasma Cu and Zn was found (P<0.05) at both days 21
and 42 of Cu supplementation. Supplemental dose of Cu-salt linearly increased (P<0.05)
plasma Cu and decreased (P<0.05) plasma Zn level but did not alter (P>0.10) other plasma
trace minerals like Mn and Fe both at days 21 and 42. Supplementation of Cu-P showed
higher (P<0.05) concentration of plasma Cu both at starter and finisher period. Reduction
of plasma Zn concentration was more for CuSO4 compared to Cu-P. Main effect of SBO
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233
Table 4
Concentrations of plasma lipids, blood reduced glutathione and body weight in broiler chicken supplemented with oil and two sources of Cu-salt (Cu-sulphate and
Cu-proteinate) at day 42 of Cu supplementation (each group having four replicates)
Attributes No supplemental oil in the diet Supplemental oil 40 g/kg diet DM S.E. Significance of treatment effects (P > F)
Control, 0 Source and dose of supplemental Control, 0 Source and dose of supplemental Oil Source Dose
copper mg/kg diet DM copper mg/kg diet DM
Cu-sulphate Cu-proteinate Cu-sulphate Cu-proteinate Linear Quadratic
200 400 200 400 200 400 200 400
Body weight (g/bird) 1745 1856 1933 1896 1976 1830 1887 1955 1917 1982 3.8 <0.001 <0.001 <0.001 0.202
Cholesterol (mmol/L) 3.83 3.54 3.44 3.44 3.36 3.62 3.37 3.34 3.25 3.16 0.024 <0.001 <0.001 <0.001 0.003
HDL-cholesterol (mmol/L) 2.39 2.24 2.23 2.21 2.28 2.33 2.20 2.21 2.14 2.19 0.024 0.002 0.313 <0.001 <0.001
LDL-cholesterol (mmol/L) 0.81 0.71 0.64 0.68 0.57 0.82 0.72 0.67 0.68 0.57 0.013 0.272 <0.001 <0.001 0.178
LDL/HDL 0.34 0.32 0.29 0.31 0.25 0.35 0.33 0.30 0.32 0.26 0.003 <0.001 <0.001 <0.001 0.232
Triglyceride (mmol/L) 0.88 0.86 0.87 0.86 0.87 0.78 0.79 0.77 0.80 0.78 0.005 <0.001 0.124 0.685 0.915
Total lipid (mg/dL) 368 368 365 364 367 364 366 369 366 364 1.7 0.339 0.092 0.839 0.853
Phospholipids (mmol/L) 138 137 135 136 133 152 146 142 144 140 0.9 <0.001 0.017 0.002 0.793
VLDL-cholesterol (mmol/L) 0.11 0.11 0.10 0.11 0.10 0.08 0.07 0.07 0.07 0.07 0.005 0.015 0.988 0.803 0.998
Blood GSH (mg/100 mL) 64 61 60 59 56 68 68 64 66 62 0.3 <0.001 <0.001 <0.001 0.991

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M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233
Table 5
Concentration of plasma trace minerals (␮g/mL) in broiler chicken supplemented with two sources of Cu-salt (Cu-sulphate and Cu-proteinate) at day 21 and with SBO
and two sources of Cu-salt at day 42 of Cu supplementation (each group having four replicates)
Attributes No supplemental SBO in the diet Supplemental SBO S.E. Significance of treatment effects
40 g/kg diet DM (P > F)
Control, 0 Source and dose of supplemental Control, 0 Source and dose of supplemental SBO Source Dose
copper mg/kg diet DM copper mg/kg diet DM
Cu-sulphate Cu-proteinate Cu-sulphate Cu-proteinate Linear Quadratic
200 400 200 400 200 400 200 400
Day 21
Copper 0.29 0.36 0.41 0.39 0.45 0.041 0.042 0.015 0.512
Zinc 2.4 2.2 2.0 2.3 2.2 0.11 0.025 0.026 0.986
Manganese 0.09 0.10 0.09 0.10 0.10 0.016 0.652 0.869 0.452
Iron 1.83 1.79 1.91 1.85 1.80 0.148 0.888 0.892 0.870
Day 42
Copper 0.30 0.38 0.46 0.41 0.52 0.27 0.37 0.45 0.40 0.51 0.018 0.133 0.002 <0.001 0.786
Zinc 2.36 2.28 2.16 2.30 2.20 2.30 2.26 2.17 2.30 2.21 0.021 0.189 0.026 <0.001 0.129
Manganese 0.10 0.09 0.09 0.10 0.10 0.96 0.97 0.99 0.95 0.93 0.012 0.142 0.733 0.713 0.892
Iron 1.87 1.91 1.77 1.82 1.89 1.80 1.84 1.94 1.86 1.94 0.056 0.708 0.660 0.334 0.954
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 223

was not found to alter (P>0.10) the plasma trace mineral concentration under study during
finisher period.

3.5. Effect on major mineral balance

Metabolism trial of days 21 and 42 revealed no effect (P>0.10) of Cu-salt and dose on
Ca, P and Mg intake (mg/day) and retention coefficient and the mean retention coefficient
of Ca, P and Mg were 0.62, 0.72, 0.35 and 0.64, 0.71, 0.35, respectively, during days 21
and 42 of experiment.

3.6. Effect on trace mineral balance

Effect of SBO and two sources of Cu-salt on trace minerals (Cu, Zn, Mn, and Fe) balance
at days 21 and 42 were presented in Tables 6 and 7, respectively.

3.6.1. Starter period

Intake of Cu increased linearly (P<0.01) with the dose of Cu-salt, which was expected.
Broiler chicken fed diet supplemented with Cu-P showed highest (P<0.01) retained (mg/day)
Cu compared to inorganic Cu, i.e. CuSO4 . The retention coefficient of Cu was reduced
(P<0.01) from 0.28 (control) to 0.18 (400 mg/kg CuSO4 ) or 0.21 (400 mg/kg Cu-P) when
dose of Cu increased in both the sources. Dose of Cu-salt showed linear response at both
day 21 (P<0.01) and day 42 (P<0.05). Amount of retained Cu in 400 mg/kg CuSO4 and
Cu-P were 23.79 and 29.31 times higher than control which established better availability of
Cu-P than CuSO4 . Supplementation of Cu-salt decreased (P<0.05) Zn retention coefficient.

Table 6
Trace minerals intake (mg/day) and retention (coefficient) of broiler chicken supplemented with two sources of
Cu-salt (Cu-sulphate and Cu-proteinate) at day 21 of Cu supplementation (each group having four replicates)
Attributes Control, 0 Source and dose of supplemental S.E. Significance of treatment
copper mg/kg diet DM effects (P > F)
Copper sulphate Copper proteinate Source Dose
200 400 200 400 Linear Quadratic
Copper
Intake 0.66 13.42 25.78 13.14 25.99 0.137 0.808 <0.001 0.953
Retention 0.28 0.21 0.18 0.24 0.21 0.002 0.0002 <0.001 0.042
Zinc
Intake 5.10 5.20 5.14 5.11 5.17 0.049 0.492 0.303 0.564
Retention 0.30 0.22 0.15 0.24 0.21 0.003 <0.001 <0.001 0.480
Manganese
Intake 6.86 6.99 6.92 6.88 6.96 0.059 0.492 0.303 0.564
Retention 0.21 0.23 0.25 0.23 0.25 0.002 0.345 0.257 0.421
Iron
Intake 6.86 6.99 6.92 6.88 6.96 0.059 0.492 0.303 0.564
Retention 0.20 0.21 0.22 0.23 0.24 0.002 <0.001 <0.001 0.092
 224
M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233
Table 7
Trace minerals intake (mg/day) and retention (coefficient) of broiler chicken supplemented with SBO and two sources of Cu-salt (Cu-sulphate and Cu-proteinate) at day
42 of Cu supplementation (each group having four replicates)
Attributes No supplemental SBO in Supplemental SBO S.E. Significance of treatment effects
the diet 40 g/kg diet DM (P > F)
Control, 0 Source and dose of supplemental Control, 0 Source and dose of supplemental SBO Source Dose
copper mg/kg diet DM copper mg/kg diet DM
Cu-sulphate Cu-proteinate Cu-sulphate Cu-proteinate Linear Quadratic
200 400 200 400 200 400 200 400
Copper
Intake 1.7 28.9 56.0 30.2 58.5 1.6 27.3 58.8 28.8 58.0 0.22 0.346 <0.001 <0.001 0.024
Retention 0.32 0.29 0.25 0.30 0.27 0.27 0.25 0.23 0.29 0.28 0.014 0.008 0.008 0.036 0.484
Zinc
Intake 12.9 11.3 11.2 11.7 12.0 12.4 10.5 11.7 11.2 11.5 0.07 0.125 0.116 0.253 0.432
Retention 0.26 0.23 0.20 0.24 0.22 0.21 0.20 0.18 0.21 0.19 0.011 0.000 0.041 0.006 0.771
Manganese
Intake 18.3 16.1 16.0 16.7 17.1 18.4 15.6 17.3 16.5 17.1 0.10 0.175 0.185 0.118 0.215
Retention 0.19 0.16 0.15 0.16 0.15 0.16 0.15 0.14 0.15 0.15 0.003 0.0002 0.640 0.206 0.211
Iron
Intake 49 43 42 45 46 48 41 45 43 45 0.3 0.113 0.125 0.356 0.226
Retention 0.12 0.14 0.15 0.15 0.14 0.11 0.13 0.14 0.14 0.14 0.411 0.089 0.253 0.312 0.281
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 225

Effect of dose of Cu-salt on retention of Zn was linear (P<0.01). The rate of reduction on Zn
retention by Cu was more in CuSO4 than Cu-P. Reduction of Zn by 400 mg/kg CuSO4 and
Cu-P were 50.3 and 21.9%, respectively, compared to control. Supplemental Cu source and
dose had no effect (P>0.10) on Mn balance. Cu-P supplementation showed an increasing
trend in Fe retention coefficient (P<0.10) compared to CuSO4 . Increase in dose of Cu-salt
showed an increasing trend (P<0.10) in Fe retention.

3.6.2. Finisher period

Intake of Cu increased linearly (P<0.01) with the dose of Cu supplement, whereas intake
of other trace minerals was unaffected. Effect of addition of SBO reduced (P<0.05) retention
coefficient of Cu and Zn but showed decreasing trend (P<0.10) for Mn and Fe. Comparison
between the two controls (with or without oil), where no Cu was supplemented showed
18.5, 19.8, 14.4 and 12.4% reduction in of Cu, Zn, Mn and Fe, respectively. Effect of
supplemental Cu-P improved (P<0.01) Cu retention compared to CuSO4 . Dose of Cu-salt
had both linear and quadratic response (P<0.01) for Cu retention. The Zn retention was
decreased (P<0.05) by addition of both SBO and source of Cu-salt in the finisher diet.
The rate of decrease (P<0.05) was less in Cu-P than CuSO4 . Dose of Cu-salt linearly
decreased (P<0.05) Zn retention. Mn retention was unaffected (P>0.10) by variation of
source and dose of Cu-salt. Cu-P supplementation had an increasing trend (P<0.10) in Fe
retention.

3.7. Effect on carcass traits

Body weight, carcass weight and abdominal fat content of broiler chickens at days 21
and 42 were presented in Table 8. At day 21 body weight of birds at the time of slaughter
and after slaughter (carcass weight) differed (P<0.01) with variation of source and dose of
Cu-salt. Cu-P supplemented birds showed higher (P<0.01) carcass weight (CW) compared
to CuSO4 supplementation. CW was increased both linearly and quadratically (P<0.05)
with the dose of Cu-salt. Proportion of liver, thigh meat, breast meat, breast and thigh meat,
drumstick, and back meat of carcass weight were not varied (P>0.10) with the variation
of source and dose of Cu-salt (not shown in table) whereas, proportion of abdominal fat
was decreased (P<0.01) with the increased dose of Cu-salt irrespective of source. Cu-
P showed a decreasing trend (P<0.10) in the proportion of abdominal fat compared to
CuSO4 .
BW and carcass weight (CW) of experimental birds in slaughter studies differed (P<0.01)
with the incorporation of SBO and variation in dose of Cu-salt in the finisher diet. Addition
of oil increased (P<0.01) the weight (BW and CW) compared to without oil. Cu-P produced
higher CW compared to CuSO4 . Dose of supplemental Cu-salt linearly increased (P<0.01)
CW of birds. The dressing % across the treatments was similar both at days 21 and 42.
Proportion of liver, thigh meat, breast meat, breast and thigh meat, drumstick and breast
meat of carcass weight were similar (P>0.10) among the treatments (not shown in table).
Proportion of abdominal fat content increased (P<0.05) by the addition of oil and decreased
(P<0.01) as the dose of Cu-salt increased. Decreasing trend (P<0.10) in abdominal fat
content was found in Cu-P compared to CuSO4 .
 226
M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233
Table 8
Body weight, carcass weight and abdominal fat content of broiler chicken supplemented with two sources of Cu-salt (Cu-sulphate and Cu-proteinate) at day 21 and with
oil and two sources of Cu-salt at day 42 of Cu supplementation (each group having four replicates)
Attributes No supplemental oil in the diet Supplemental oil 40 g/kg diet DM S.E. Significance of treatment effects
(P > F)
Control, 0 Source and dose of supplemental copper Control, 0 Source and dose of supplemental copper Oil Source Dose
mg/kg diet DM mg/kg diet DM
Copper sulphate Copper proteinate Copper sulphate Copper proteinate Linear Quadratic

200 400 200 400 200 400 200 400

Day 21
Body weight (g/bird) 552 576 583 581 596 1.6 <0.001 <0.001 0.005
Carcass weight (g/bird) 343 359 362 361 372 0.229 <0.001 0.019 0.062
Abdominal fat (% 1.0 0.9 0.9 0.9 0.9 1.71 0.007 0.932 0.879
carcass weight)
Day 42
Body weight (g/bird) 1745 1856 1933 1896 1976 183 1887 1955 1917 1983 3.8 <0.001 <0.001 <0.001 0.202
Carcass weight (g/bird) 1279 1359 1417 1390 1447 1342 1382 1432 1404 1452 4.9 <0.001 <0.001 <0.001 0.230
Abdominal fat (% 1.25 1.21 1.13 1.19 1.10 1.30 1.24 1.16 1.21 1.13 0.021 0.021 0.082 <0.001 0.584
carcass weight)
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 227

4. Discussion

4.1. Plasma lipids

In experiment with broiler chicken, Bakilli et al. (1995) and Pesti and Bakalli (1996)
showed supplemental Cu at pharmacological levels in the diet decreased plasma and breast
muscle cholesterol. Paik et al. (1999) reported reduced plasma cholesterol from high level
of inorganic Cu but no reduction from organic Cu–methionine chelate even from breast
muscle of broiler chicken. The present study showed Cu supplementation reduced plasma
cholesterol, as in the reports of Ankari et al. (1998) and Pesti and Bakalli (1998). Studies
with broiler have shown a similar effect of reducing plasma and meat cholesterol concen-
tration (Konjufca et al., 1997; Skrivan et al., 2000). Kim et al. (1992) demonstrated that
Cu deficiency hypercholesterolemia by elevating hepatic GSH (reduced glutathione) levels
and changing the GSH:GSSG (oxidized glutathione) ratio, which decreased the activity of
the 3-hydroxy-3-methylglutaryl Co-A (HMG-CoA) reductase. If this mechanism is func-
tional in Cu deficiency, we expect that supplemental Cu would decrease the GSH level
and subsequently increase the HMG-CoA reductase activity. In the present investigation
the blood reduced GSH was decreased with Cu supplementation. Glutathione is known to
regulate cholesterol biosynthesis through the stimulation of HMG-CoA reductase (Vaisala
and Kurup, 1987; Konjufca et al., 1997), the key enzyme of cholesterol biosynthesis and
ultimately reduce plasma cholesterol concentration. Reduction of plasma cholesterol on
days 21 and 42 by supplementing Cu with different dose can be justified by the above
mentioned pathway.
In the present study, addition of SBO reduced plasma cholesterol level. Qureshi et al.
(1983) reported a correlation between HMG-CoA reductase activity and either total or LDL-
C in chicken but not with the HDL-C. The complete inhibition of cholesterol synthesis as
proposed by Goldstein and Brown (1990) requires two regulators, i.e. cholesterol from
LDL and a non-sterol product derived from mevalonate both of which modulate HMG-
CoA reductase activity. It may be hypothesized that SBO supplementation may regulate a
non-sterol product to reduce plasma cholesterol level.
The present study revealed that supplementation of Cu increased HDL-C day 21. In
previous studies, Cu supplementation to broiler chicken showed reduction in VLDL and
increased HDL-C concentration (Lien et al., 2001, 2004) which is consistent with the
present findings, whereas Chowdhury et al. (2004) could not found effect of Cu-Met
chelate on HDL-C concentration. Paik et al. (1999) observed a little increase in HDL-C
concentration with Cu-Met chelate at 125 and 250 mg/kg supplementation level of Cu but
those data were beyond statistical significance. It was found that in Cu deficient animals,
total cholesterol and free cholesterol levels were elevated and were associated with low
concentration of HDL-C (Lei, 1983; Lefevre et al., 1986). Lefevre et al. (1986) suggested
that a primary biochemical lesion underlying the hypercholesterolemia is a reduction in
hepatic HDL binding site which would result in a slower turn over of HDL and lead to
an accumulation of apo-E rich HDL. Therefore it may be inferred that supplementation of
Cu-salt may increase hepatic binding sites which would be helpful for better utilization of
cholesterol. Cu supplementation from two different sources of Cu-salt reduced LDL-C but
had no effect on VLDL-C concentration which was indicative of synthesis of favourable
228 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233

cholesterol (HDL-C) from LDL-C by modulating HMG-CoA reductase activity (Goldstein

and Brown, 1990) by Cu. Supplementation of SBO reduced HDL-C concentration, whereas
LDL-C increased with SBO supplementation and subsequently decreased when Cu is
used with SBO. VLDL-C decreased with oil supplementation but supplemental Cu has
no effect. Crespo and Esteve-Garcia (2003) reported reduction of VLDL-C concentration
in broiler fed sunflower and linseed oil in the fasted state which corroborate the present
findings.
The effect of Cu on triglyceride concentration was found at day 21 but supplemen-
tal Cu could not affect triglyceride concentration at day 42 when SBO was used with
Cu. SBO supplementation significantly reduced plasma triglyceride. Chowdhury et al.
(2004) found plasma triglyceride concentration increased quadratically when dietary level
of Cu-Met chelate was between 100 and 150 mg/kg. Paik et al. (1999) obtained reduc-
tion of triglyceride at 125 mg/kg dietary level of Cu but little difference in 250 mg/kg
Cu supplementation. Plasma triglyceride concentration was also decreased in broiler fed
Cu from inorganic sources at 250 mg/kg level (Bakilli et al., 1995). In the present find-
ings organic Cu showed better reduction in plasma triglyceride than inorganic source.
When fat was used in the diet reduction of triglyceride concentration by fat was so high
that effect of Cu in decreasing triglyceride was not found. The decrease in triglyceride
formation from PUFA is likely a result of PUFA regulation of enzymes (ATP-citrate
lyase, acyl CoA carboxylase and fatty acid synthase) involved in triglyceride synthe-
sis (Iritani et al., 1998; Mashek et al., 2002). These enzymes are decreased by diets
high in PUFA (Geelan et al., 1995; Fremont and Gozzelino, 1996; Cha et al., 1998).
In the present study soybean oil was used which is rich in PUFA and reduce plasma
triglyceride.
Effect of Cu on phospholipids was found at both days 21 and 42 with decrease in
concentration. But SBO supplementation increased this value in chickens. The effect of
Cu on phospholipids concentration may be explained by the reduction of reduced GSH
concentration which was found in the present study. Amer and Elliot (1973) found that
the addition of Cu markedly increased the phospholipids concentration and proportion of
unsaturated fatty acids in pigs fed on Cu supplement diet which contradicts the present
findings. Though supplementation of Cu and SBO alter the plasma lipid profiles but plasma
total lipid was unchanged throughout the experimental period.

4.2. Haemato-biochemical profile

The influence of supplemental Cu-salt with different dose to broiler chicken could not
produce any significant alteration in plasma glucose and proteins and enzyme concentra-
tions. Concentrations of plasma glucose, proteins and enzymes levels were in normal range
which, signify that the animals were apparently healthy throughout the experimental period.
Levels of plasma AST, ALT, ALP, GGT and LDH were positively correlated with increased
Cu intake and indicative of hepatic damage starting with 300 mg Cu/head/day in goat kids
(Haenlein, 2004). Thompson and Todd (1974) reported that serum AST activity begin to rise
during the prehaemolytic period. So it can be conclude that supplementation of Cu-salt up
to 400 mg/kg could not able to produce any dystrophy in hepatic or other tissues containing
these enzymes during the feeding trial. Thus no clinical symptom of toxicity was developed
 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233 229

in these birds. Rabiansky et al. (1999) also evaluated the effect of Cu-lysine and CuSO4
source for heifers and found no effect of Cu source and concentration on haemoglobin levels
which is consistent with the present findings.

4.3. Plasma mineral

Addition of Cu increases plasma Cu concentration (Luo and Dove, 1996; Castell and
Bowland, 1968; Dove and Haydon, 1992; Zhou et al., 1994a,b; Gipp et al., 1973, 1974)
indicating that dietary Cu levels were reflected in plasma Cu concentration which is
consistent with the present findings. Gipp et al. (1974) reported a reduction of plasma
Zn by supplementing dietary Cu which corroborates the present findings. The lack of
high dietary Cu treatment effect on plasma Zn concentration was noted by Roof and
Mahan (1982) which contradicts the present findings. Supplemental Cu in the organic
form including Cu-lysine, Cu-methionoine and Cu-P was absorbed equal to or greater than
that from cupric sulphate (Kincaid et al., 1986; Baker et al., 1991) increase plasma Cu
concentration.

4.4. Mineral balance

Major minerals (Ca, P, and Mg) balance was not affected by the dietary Cu-salt and
SBO supplementation both at days 21 and 42. This finding can be compared with the
findings of Dove (1995) who found no effect on Ca, P and Mg retention by addition of
fat or Cu to the diet of pigs. Trace minerals complexed with organic molecules have been
implied to be more bioavailable than inorganic trace minerals (Brown and Zeringue, 1994).
Some researchers (Nockels et al., 1993; Rabiansky et al., 1999) have indicated that Cu-
lysine may be a more available source of Cu than Cu-sulphate in cattle. The physiological
advantage afforded by organic Cu compounds may be due to unique coordination chemistry
of Cu, which permits the formation of highly soluble, chemically stable products that resist
interaction with antagonists in the gut (Brown and Zeringue, 1994). Kegley and Spears
(1994) also reported that Cu status of calves fed Cu-sulphate did not differ from calves fed
Cu-lysine. Nockels et al. (1993) reported greater apparent Cu absorption and retention of
organic Cu compounds compared with Cu-sulphate in calves following an 18 days stress
period that included ACTH injection and mineral restriction. In rats it has been found that
Cu and Zn mutually hamper absorption of each other (Van Campen and Scaife, 1967; Van
Campen, 1966). In the present study supplementation of Cu also reduced the retention
of Zn.
Hart et al. (1928) reported that an anemia caused by the defective utilization of Fe, was the
result of Cu deficiency and that Cu is required for Fe metabolism. Defective Fe metabolism
in Cu deficient animals is thought to be due to low levels of ferroxidase activity in the blood.
Transferrin transports Fe in blood and binds only to Fe(III). Iron is released from the liver
and absorbed as Fe(II). Frieden and Hsieh (1976) proposed that the oxidation of Fe(II) to
Fe(III) is catalyzed by ferroxidase enzymes, which contain Cu (Prohaska, 1991). Although
plasma ferroxidase activity was not measured in the study, it is possible that elevated levels
of dietary Cu increased ferroxidase activity and caused a shift in equilibrium that increased
Fe absorption.
230 M.K. Mondal et al. / Animal Feed Science and Technology 139 (2007) 212–233

4.5. Carcass traits

In general, body fat accumulation may be considered the net result of balance among
dietary absorbed fat, endogenous fat synthesis (lipogenesis) and fat catabolism via ␤-
oxidation (lipolysis). Lower body fat deposition may be attributed to increased fat catabolism
and diminished endogenous fatty acid synthesis or to both processes. Effect of Cu on abdom-
inal fat content can be explained by the way that Cu may alter lipogenesis or lipolysis in the
body and decreased abdominal fat whereas; supplementation of SBO increased abdominal
fat content. Abdominal fat content of the birds of the current experiment was quite low,
probably due to the low ME:CP ratio used in diets.

5. Conclusion

The data from this feeding trial indicate that supplemental Cu decreased plasma choles-
terol, LDL-C and increased HDL-C in broiler chicken even in presence of SBO (40 g/kg
diet). Supplemental Cu-salt increased retained Cu and decreased Zn but did not affect major
mineral balance. Cu-P is more bioavaliable than CuSO4 which is evident from plasma min-
eral and mineral balance study. Cu-salt supplementation at the rate of 400 mg/kg diet was
not found to be toxic to the broiler birds during this feeding trial. Dose and source of Cu-
salt had an important role in plasma lipids, plasma minerals and mineral balance of broiler
chickens.

6. Implication

Cu supplementation at pharmacological level, i.e. at 200 mg/kg diet may alter choles-
terol and lipid metabolism in broiler chickens. The potential of reducing cholesterol in
broiler meat produced for human consumption has many health benefits. Addition of Cu
at pharmacological level can reduce plasma cholesterol even in feeding soybean oil and
may subsequently reduce meat cholesterol. As soybean oil is most commonly used energy
source in finisher diet of broiler birds in Indian condition, so Cu may be used with soybean
oil to reduce health hazards produce by consuming soybean oil.

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