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ORIGINAL ARTICLE
Summary
This study was designed to evaluate the effects of dietary inclusion of zinc-methionine (ZnMet) as a replacement for conventional inorganic zinc sources on performance, tissue zinc accumulation and some plasma indices in broiler chicks. A total of 450-day-old Ross male broiler chicks were randomly assigned to five pen
replicates of nine experimental diets. Dietary treatments consisted of two basal diets supplemented with
40 mg/kg added Zn as feed-grade Zn sulphate or Zn oxide in which, Zn was replaced with that supplied from
ZnMet complex by 25, 50, 75 or 100%. At 42 days of age, three randomly selected birds from each pen were
bled to measure plasma metabolites; then, the chicks were slaughtered to evaluate carcass characteristics.
Results showed that dietary treatments affected (p < 0.05) feed intake during the starter period, and chicks on
Zn oxide diets consumed more feed than sulphate counterparts. Furthermore, dietary substitution of inorganic
Zn sources by ZnMet caused improvements (p < 0.01) in body weight gain during all experimental periods.
Dietary supplementation of ZnMet improved feed conversion efficiency during 121 and 142, but not in 21
42 days of age. Complete replacement of inorganic Zn by that supplied from ZnMet caused an increase
(p < 0.05) in relative liver weight. Similarly, dietary inclusion of ZnMet increased breast meat and carcass
yields and reduced abdominal fat percentage (p < 0.05). Incremental levels of ZnMet increased (p < 0.05) zinc
concentrations in liver and thymus, and the highest zinc accumulations were seen in 100% ZnMet-supplemented birds. Interestingly, introduction of ZnMet into the diets partially in place of inorganic sources resulted
in decreases (p < 0.01) in plasma uric acid and triglycerides concentrations. The present findings indicated that
dietary ZnMet inclusion in replacement of inorganic sources in addition to improving growth performance,
reduced plasma uric acid and triglycerides concentrations, consequently decreased abdominal fat pad and
increased carcass meat yield.
Keywords broiler chicks, organically bound minerals, zinc-methionine, plasma triglycerides, uric acid, carcass characteristics
Correspondence R. Jahanian, Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan 84156-83111, Iran.
Tel: +98 311 391 3511; Fax: +98 311 391 3501; E-mail: r.jahanian@cc.iut.ac.ir
Received: 17 February 2013; accepted: 13 May 2014
Introduction
Zinc is known to be essential for the life of all organisms (Vallee and Auld, 1990; Coleman, 1992). The
first zinc metalloenzyme that was discovered in 1940
is carbonic anhydrase II (Keilin and Mann, 1940).
Since then, >300 zinc enzymes covering all six classes
of enzymes and in different species of all phyla have
been discovered (Vallee and Auld, 1990; Christianson,
1991; Coleman, 1992). In most cases, zinc ion is an
essential cofactor for the observed biological function
of these metalloenzymes. Furthermore, the biological
functions of zinc, which are versatile and observed in
50
Journal of Animal Physiology and Animal Nutrition 99 (2015) 5058 2014 Blackwell Verlag GmbH
dietary Zn are commonly added to reduce the possibility of zinc deficiency under commercial conditions
because of the poor availability of Zn in plant feed
ingredients (Fordyce et al., 1987). This common
practice, however, has generated yet another concern:
zinc accumulation in soil and surface water, and its
transport from poultry and livestock manure to croplands. Although environmental concerns have
focused on nitrogen and phosphorus contamination
of soils with poultry manure, some geographic regions
are becoming vulnerable to heavy metal pollution
such as Zn (Burrell et al., 2004). Broiler chicks that
are fed diets containing high amounts of Zn potentially produce faecal wastes with higher zinc contents
and cause environmental contamination with this element. Why zinc is supplemented to broiler diets in
such high concentrations may be, in part, due to the
poor availability of zinc in plant feed ingredients and
conventional inorganic zinc sources (such as zinc sulphates and oxides), caused by the binding of Zn by
phytate (Ellis et al., 1982; Fordyce et al., 1987). In
addition, much natural feedstuffs are marginally deficient in zinc (Baker and Ammerman, 1997; Cao et al.,
2002).
Some researchers (Spears, 1989; Wedekind et al.,
1992; Sandoval et al., 1997b; Jahanian et al., 2008a,
b) have reported a greater bioavailability for organic
Zn sources than that observed for inorganic forms
including Zn oxide and Zn sulphate; consequently,
organic sources of the element have been used with
increasing frequency by the feed industry during the
recent years. In their study with broiler chicks, Nassiri
Moghaddam and Jahanian (2009) observed that diet
fortification by zinc-methionine (ZnMet) complex in
place of inorganic Zn sources increased antibody production titres against Newcastle and infectious bronchitis disease viruses. Such event was observed when
ZnMet or zinc-lysine were fed to broilers (Jahanian
and Yaghoubi, 2010), indicating higher bioavailability
values of organic zinc sources. An enhanced bioavailability of a mineral source could reduce the amount of
mineral that should be added to the diet to meet the
mineral nutritional requirements, which in turn,
would reduce the amount of mineral excreted to the
environment (Cheng et al., 1998).
Despite the advantages resulting from dietary supplementation with organic mineral sources, contradictory reports and variable bioavailability values have
been reported with the trace mineral chelates and
complexes such as ZnMet, indicating no advantage
from the use of organic zinc forms (Ammerman et al.,
1995). In addition, there is no literature cited that
examines the effects of gradual substitution of
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH
51
This study was carried out at the Poultry Research Station of Isfahan University of Technology (Isfahan,
Iran), and all experimental procedures were approved
by Isfahan University of Technology Animal Care and
Use Committee. A total of 450-day-old, cloacal sexed,
male Ross 9 Ross broiler chicks were allocated to the
five pen replicates (10 birds each) for each of the nine
dietary treatments in a completely randomized design.
On day 1 post-hatching after an overnight fast, chicks
were weighed and assigned to the experimental
groups so that each group had a similar mean initial
weight and weight distribution. Dietary treatments
included the two basal diets supplemented with
40 mg/kg added Zn as feed-grade Zn sulphate or Zn
oxide (as conventional inorganic sources) in which,
Zn was replaced with that provided from Zn-methionine complex (Zinpro Corporation, Edina, MN, USA)
by 25, 50, 75 or 100%. The basal cornsoya bean meal
diets (Table 1) in both starter and grower periods were
formulated using Zn-free mineral premix and were
adequate in all nutrients (NRC, 1994) except Zn,
which was present at 25.9 and 24.2 mg/kg (by atomic
absorption analysis) in starting and growing diets,
respectively. Treatment additions were made at the
expense of inert filler (Zn-free washed builders sand)
so that, all of the diets were calculated and/ or analysed to contain equal concentrations of metabolizable
energy, crude protein and other nutrients. Chicks
were allowed ad libitum access to the experimental
diets and to tap water containing no detectable Zn
throughout the duration of the study. Feed and water
were provided using plastic equipments to minimize
environmental Zn contamination. Chicks were maintained on a 23 L: 1 D lighting regimen in the floor
pens in a thermostatically controlled room. Temperature was set at 33 C during the first week of age;
since then, reduced by 3 C/ week until the birds were
4 weeks old. At 3 and 6 weeks of age, feed consumption and body weights were recorded by pen basis
after 4 h of feed withdrawal, and mortality was
recorded daily to correct for feed: gain data. Three ran-
Starter (121
days of age)
Grower (2242
days of age)
54.31
38.41
3.00
1.85
1.22
0.40
0.25
0.25
0.16
0.15
62.01
31.27
0.75
2.25
1.35
1.32
0.30
0.25
0.25
0.10
0.15
3000
21.57
5.99
0.49
0.84
1.18
0.84
0.32
1.40
0.94
0.43
0.17
25.9
3030
18.94
6.26
0.40
0.72
1.01
0.74
0.27
1.21
0.85
0.34
0.14
24.2
acid (Table 5). Similarly, dietary substitution of inorganic zinc sources with the organically Zn compound
resulted in a decrease (p < 0.01) in plasma triglycerides content. Among blood parameters, only plasma
triglycerides was affected by the level of 100% of three
Zn sources, with the least triglyceride concentrations
assigned to the birds fed on 100% ZnMet-containing
diets followed by those on sulphate diets.
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH
53
Discussion
As noticed in Table 2, complete substitution of ZnMet
for inorganic Zn sources caused an increase (p < 0.05)
in feed intake during entire trial period (142 days of
age). On the other hand, the least feed intakes
assigned to the sulphate-supplemented groups. Similar
to our findings, Sandoval et al. (1997a) and Jahanian
et al. (2008a) showed that the chicks fed on sulphatesupplemented diets consumed less feed than those fed
other Zn sources (irrespective from that organic or
inorganic sources were fed). It seems that a part of
feed intake depression by Zn sulphate-containing diets
may be related to higher water solubility of sulphates.
The exact reason for this reduction, however, remains
unclear. The best weight gains allotted to the chicks
fed on diets containing at least 50% Zn from the
ZnMet complex (apart from that the remaining proportion provided from Zn sulphate or Zn oxide). A
greater bioefficacy of ZnMet relative to sulphates or
oxides could provide more Zn for absorption, consequently improve growth performance if Zn is potentially bound by fibre and phytates in basal diets
(Edwards and Baker, 1999; Jahanian et al., 2008a,b).
In production animals, recently, interest in using
organic mineral complexes as mineral sources has
increased because of the reported potential of their
higher bioavailability than inorganic sources (Hahn
and Baker, 1993; Schell and Kornegay, 1996; Batal
et al., 2001). The higher absorption capacity of ZnMet
allows lower inclusion rates of zinc and makes mineral
balance in animal easier to maintain, as evidenced in
study herein. The data presented here indicate that
when a nutritional requirement for zinc is considered,
Zn sources that supplied should be specified. The
source of supplemental Zn used in a Zn requirement
assay can have just as much effect on the determined
Zn requirement as the type of basal diet used. Some
researchers (Davies et al., 1977; Baker and Halpin,
1988) have reported that components contained in
corn, soya bean meal, etc. reduce Zn bioavailability.
These feed ingredients not only contain poorly available Zn, but in many cases, they also reduce the bioavailability of supplemental Zn. Presumably, tightly
54
130.57
131.34
38.76b
40.37a
0.5133
0.4930
0.0746
1.6042
0.0085
0.1389
0.0783
0.6706
0.0619
0.1402
0.0267
0.8361
0.1461
84.66
85.86
84.85
86.32
85.99
86.27
85.97
83.00
84.58
86.01
85.06
142 days
0.8869
0.0047
0.0010
0.7592
0.0040
25.59
25.69
23.26d
25.28bcd
26.70abc
27.51a
27.26ab
23.98d
24.89cd
26.46abc
27.03ab
121 days
0.5548
0.0018
0.1036
0.9721
0.0247
66.87
67.32
65.84ab
66.65ab
68.12a
68.67a
68.33a
64.49b
66.48ab
68.35a
68.15a
2242 days
0.6562
0.0076
0.0061
0.7421
0.0010
46.23
46.50
44.55cd
45.97bc
47.41ab
48.09a
47.80a
44.24d
45.69cd
47.41ab
47.59ab
142 days
0.1117
0.0031
0.0445
0.0417
0.0110
1.52
1.58
1.69a
1.61ab
1.53bc
1.48c
1.50bc
1.59abc
1.55bc
1.49bc
1.45c
121 days
0.9116
0.1747
0.3735
0.0333
0.6241
1.95
1.95
1.98
1.98
1.93
1.92
1.92
1.99
1.96
1.94
1.92
2242 days
0.5958
0.0158
0.1364
0.0331
0.1479
1.83
1.85
1.90
1.88
1.81
1.79
1.80
1.88
1.85
1.81
1.79
142 days
ad
Means with no common superscripts within each column are significantly (p < 0.05) different.
*The level (%) of total dietary added Zn, which supplied by zinc sulphate or zinc oxide; The final zinc concentrations (basal diet + supplemental Zn, 40 mg/kg) in experimental diets were 66.1, 65.7, 65.4,
66.2, 65.8, 65.6, 66.4, 65.5 and 65.7 mg/kg in starter diets in order that dietary treatments listed in table (by analysis). The respective values were determined to be 64.5, 63.7, 64.1, 63.9, 64.2, 63.4,
63.8, 64.0 and 63.7 mg/kg in grower diets.
The level (%) of total dietary added Zn, which provided by ZnMet complex.
Standard error of means.
0.7049
0.0252
p-values
130.43
131.89
131.08
131.95
130.99
39.26abc
40.75ab
40.89a
40.59ab
40.95a
25
50
75
100
128.02
130.57
132.71
130.98
2242 days
37.98c
38.59bc
39.31abc
39.14abc
121 days
25
50
75
ZnMet
Treatments
Contrasts
Sulphate vs. oxide
Replacement level
100% of each source
Pooled SEM
Zn sulphate
Zn oxide
Zn sulphate
100
75
50
25
Zn oxide
100
75
50
25
Source and
level of
inorganic Zn*
Table 2 Effects of dietary substitution (% of Zn supplied) of inorganic zinc sources by zinc-methionine (ZnMet) on performance parameters during 142 days of age
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH
Table 3 Effects of dietary substitution (% of Zn supplied) of inorganic zinc sources by zinc-methionine (ZnMet) on carcass characteristics of 42 daysaged broiler chicks (% of live body weight)
Source and level of
inorganic Zn*
Zn sulphate
100
75
50
25
Zn oxide
100
75
50
25
Zn sulphate
Zn oxide
ZnMet
Liver
Pancreas
Breast
Thigh
Carcass
Abdominal fat
25
50
75
1.97
2.03
2.14
2.21
0.22
0.23
0.25
0.23
20.25b
20.42b
20.92ab
21.02ab
21.05
19.80
19.57
19.92
70.13c
70.87abc
70.64bc
71.59ab
1.84ab
1.78abc
1.72abc
1.58c
25
50
75
100
2.10
2.17
2.01
2.15
2.27
0.25
0.22
0.22
0.24
0.25
21.07ab
20.66b
21.15ab
21.73a
21.09ab
19.39
19.76
19.61
20.13
20.34
71.30abc
71.09abc
71.74ab
72.16a
71.93ab
1.93a
1.94a
1.83abc
1.69abc
1.62bc
2.09
2.11
0.23
0.23
20.65b
21.15a
20.09
19.72
70.81b
71.57a
1.73
1.85
p-values
Treatments
Contrasts
Sulphate vs. oxide
Replacement level
100% of each source
Pooled SEM
0.0809
0.3194
0.0426
0.3112
0.0410
0.0281
0.7025
0.0703
0.0317
0.0690
1.0000
0.6192
0.0806
0.0120
0.0300
0.0654
0.1366
0.2830
0.2816
0.5950
0.0413
0.4508
0.0146
0.0689
0.0474
0.5216
0.0617
0.0097
0.0545
0.0787
abc
Means with no common superscripts within each column are significantly (p < 0.05) different.
*The level (%) of total dietary added Zn, which supplied by zinc sulphate or zinc oxide; The final zinc concentrations (basal diet + supplemental Zn,
40 mg/kg) in experimental diets were 66.1, 65.7, 65.4, 66.2, 65.8, 65.6, 66.4, 65.5 and 65.7 mg/kg in starter diets in order that dietary treatments listed
in table (by analysis). The respective values were determined to be 64.5, 63.7, 64.1, 63.9, 64.2, 63.4, 63.8, 64.0 and 63.7 mg/kg in grower diets.
The level (%) of total dietary added Zn, which provided by ZnMet complex.
Standard error of means.
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH
55
DOI: 10.1111/jpn.12213
ORIGINAL ARTICLE
Summary
This study was designed to evaluate the effects of dietary inclusion of zinc-methionine (ZnMet) as a replacement for conventional inorganic zinc sources on performance, tissue zinc accumulation and some plasma indices in broiler chicks. A total of 450-day-old Ross male broiler chicks were randomly assigned to five pen
replicates of nine experimental diets. Dietary treatments consisted of two basal diets supplemented with
40 mg/kg added Zn as feed-grade Zn sulphate or Zn oxide in which, Zn was replaced with that supplied from
ZnMet complex by 25, 50, 75 or 100%. At 42 days of age, three randomly selected birds from each pen were
bled to measure plasma metabolites; then, the chicks were slaughtered to evaluate carcass characteristics.
Results showed that dietary treatments affected (p < 0.05) feed intake during the starter period, and chicks on
Zn oxide diets consumed more feed than sulphate counterparts. Furthermore, dietary substitution of inorganic
Zn sources by ZnMet caused improvements (p < 0.01) in body weight gain during all experimental periods.
Dietary supplementation of ZnMet improved feed conversion efficiency during 121 and 142, but not in 21
42 days of age. Complete replacement of inorganic Zn by that supplied from ZnMet caused an increase
(p < 0.05) in relative liver weight. Similarly, dietary inclusion of ZnMet increased breast meat and carcass
yields and reduced abdominal fat percentage (p < 0.05). Incremental levels of ZnMet increased (p < 0.05) zinc
concentrations in liver and thymus, and the highest zinc accumulations were seen in 100% ZnMet-supplemented birds. Interestingly, introduction of ZnMet into the diets partially in place of inorganic sources resulted
in decreases (p < 0.01) in plasma uric acid and triglycerides concentrations. The present findings indicated that
dietary ZnMet inclusion in replacement of inorganic sources in addition to improving growth performance,
reduced plasma uric acid and triglycerides concentrations, consequently decreased abdominal fat pad and
increased carcass meat yield.
Keywords broiler chicks, organically bound minerals, zinc-methionine, plasma triglycerides, uric acid, carcass characteristics
Correspondence R. Jahanian, Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan 84156-83111, Iran.
Tel: +98 311 391 3511; Fax: +98 311 391 3501; E-mail: r.jahanian@cc.iut.ac.ir
Received: 17 February 2013; accepted: 13 May 2014
Introduction
Zinc is known to be essential for the life of all organisms (Vallee and Auld, 1990; Coleman, 1992). The
first zinc metalloenzyme that was discovered in 1940
is carbonic anhydrase II (Keilin and Mann, 1940).
Since then, >300 zinc enzymes covering all six classes
of enzymes and in different species of all phyla have
been discovered (Vallee and Auld, 1990; Christianson,
1991; Coleman, 1992). In most cases, zinc ion is an
essential cofactor for the observed biological function
of these metalloenzymes. Furthermore, the biological
functions of zinc, which are versatile and observed in
50
Journal of Animal Physiology and Animal Nutrition 99 (2015) 5058 2014 Blackwell Verlag GmbH
uric acid reduced in birds fed on >50% ZnMet-containing diets. Probably, both Zn and Met contained in
ZnMet are more bioavailable and promote protein biosynthesis, subsequently reduce blood triglycerides and
cholesterol contents.
In conclusion, the present findings show that partial
substitution of inorganic Zn by ZnMet is accompanied
with better growth performance and feed conversion
efficiency in broiler chicks. On the other hand, dietary
inclusion of ZnMet into the broiler diets could
improve carcass and breast meat yields and reduce
abdominal fat percentage. It appears that one could
achieve major or complete beneficial impacts of
ZnMet by replacing 5075% of inorganic zinc sources
with zinc-methionine complex.
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH
57
58
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH