DOI: 10.1111/jpn.

12213

ORIGINAL ARTICLE

Effects of dietary substitution of zinc-methionine for inorganic

zinc sources on growth performance, tissue zinc accumulation
and some blood parameters in broiler chicks
R. Jahanian and E. Rasouli
Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, Iran

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

many tissues, are most often associated with proteins

(McCall et al., 2000).
Zinc deficiency arrests cell proliferation, while in
multicellular organisms, it also results in abnormal differentiation and development, leading to extensive
abnormalities or syndromes (Christianson, 1991;
Coleman, 1992). Supplemental zinc is added to poultry diets to prevent the problems associated with Zn
deficiencies. In this case, National Research Council
(1994) recommends the level of 40 mg added zinc/kg
feed for broilers; however, the poultry rations are
often formulated to contain dietary zinc concentrations over 80 mg/kg. The higher concentrations of

Journal of Animal Physiology and Animal Nutrition 99 (2015) 5058 2014 Blackwell Verlag GmbH

R. Jahanian and E. Rasouli

Effect of zinc-methionine on broiler performance

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

conventional inorganic Zn sources by ZnMet on

growth performance and plasma biochemical indices
in broiler diets. Therefore, the main objectives of this
study were to evaluate the effects of partial or complete substitution of ZnMet for inorganic Zn sources
on performance, carcass characteristics, tissue zinc
accumulations and plasma variables of male broiler
chicks.

Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH

51

Materials and methods

Birds, diets and general procedures

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-

Effect of zinc-methionine on broiler performance

R. Jahanian and E. Rasouli

Table 1 Ingredient composition and chemical analysis of basal diets

during starter and grower periods (as fed basis)
Ingredients (% unless
stated otherwise)
Corn, yellow
Soya bean meal
Sunflower oil
Poultry fat
Dicalcium phosphate
Limestone
Common salt
Mineral premix*
Vitamin premix
DL-methionine
Variable
Nutrient composition
MEn (kcal/kg)
Crude protein
Ether extract
Methionine
Methionine + Cysteine
Lysine
Threonine
Tryptophan
Arginine
Calcium
Non-phytate P
Sodium
Zinc (mg/kg)

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

*Zinc-free mineral premix provided per kilogram of diet: Mn (from

MnSO4.H2O), 60 mg; Fe (from FeSO4.7H2O), 50 mg; Cu (from
CuSO4.5H2O), 6 mg; I (from Ca (IO3)2.H2O), 1 mg; and Se, 0.20 mg.
Provided per kilogram of diet: vitamin A (from vitamin A acetate),
9700 IU; cholecalciferol, 3300 IU; vitamin E (from DL-a-tocopheryl acetate), 26 IU; vitamin B12, 0.31 mg; riboflavin, 6.6 mg; niacin, 48 mg; calcium pantothenate, 35 mg; menadione (from menadione dimethylpyrimidinol), 2.5 mg; folic acid, 1.2 mg; thiamine, 3 mg; pyridoxine,
6.5 mg; biotin, 0.6 mg; choline (from choline chloride 60%), 560 mg; and
ethoxyquin, 125 mg.
Variable amounts of zinc sources and inert filler (Zn-free washed builders sand). Zinc sources were added in place of an equivalent weight of
sand.

domly selected birds from each pen were weighed

individually and bled via wing vein at 42 days of age
to measure plasma uric acid, triglycerides and cholesterol contents. These plasma variables were measured
based on the fact that Zn and Met are the vital and
limiting nutrients for protein biosynthesis, whereby
could affect lipid metabolites and uric acid (Kaim and
Schwederski, 1994; Lesson and Summers, 2001). After
that, the birds were killed by cutting the jugular vein,
allowed to bleed for approximately 2 min. Thereafter,
liver, pancreas and abdominal fat were excised and
weighed immediately. Carcass was eviscerated manu52

ally and weighed. Breast and thigh muscles were cut

(skinless) from a same location for all of the slaughtered
birds and weighed separately to measure breast and
thigh meat yields. Liver and thymus were removed,
pooled by replicate and frozen at 20 C until farther
analysis for zinc concentration. Also, plasma samples
were pooled and saved for mineral analysis.
Chemical analysis

Prior to formulating the diets, all of the ingredients

used were analysed for crude protein (code 976.06),
ether extract (code 920.39), crude fibre (code 978.10)
and ash (code 942.05) contents according to the standard procedures of AOAC (1995); then, the metabolizable energy values of feed ingredients were estimated
using National Research Council (1994) recommended formula. Zinc concentrations in Zn sources,
feedstuffs, plasma and tissues were determined by
atomic absorption spectrophotometry (Perkin Elmer,
Precisely Analyst 200, Atomic Absorption Spectrophotometer). Samples of feedstuffs, Zn sources and tissues
were dried at 105 C for 8 h, dry-ashed at 550 C
overnight, then solubilized in HCl and filtered
through Whatman 42 filter paper. Drinking water was
concentrated 10-fold by evaporation on a hot plate.
The samples were diluted appropriately with deionized water to determine Zn content (Anonymous,
1982). Plasma uric acid, triglycerides and cholesterol
were analysed with a biochemical analyzer (ERBA
CHEM-5; Beijing Biochemical Instrument Company,
Beijing, China) using the standard kits (Pars Azmoun,
Tehran, Iran).
Statistical analysis

All of the data were analysed using General Linear

Model procedures of SAS software (SAS Institute,
1999). Pen was the experimental unit for all measurements. For the carcass and plasma measurements
data, the means of three birds were pooled and participated in the analysis of variance. Treatment means
were compared using Tukeys tests (Steel et al., 1997)
at p < 0.05 significant level. Contrast comparisons
were made among the treatment means to compare
sulphate- and oxide-supplemented diets, replacement
levels and level of 100% for all three Zn sources.
Results
The zinc sources used in study here (zinc sulphate,
zinc oxide and ZnMet) were determined to be containing 32.64, 75.73 and 9.31% zinc as fed basis,
Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH

Effect of zinc-methionine on broiler performance

R. Jahanian and E. Rasouli

respectively. Only inorganic zinc sources (Zn sulphate

and Zn oxide) contained Fe ions, which determined
to be 1486 and 29650 mg/kg respectively. Copper
content of Zn sulphate and Zn oxide was 27 and
609 mg/kg, respectively, and ZnMet complex had no
detectable copper. The manganese was also contained
only in inorganic zinc sources so that, Zn sulphate
and Zn oxide contained 1572 and 492 mg Mn2+/kg
respectively.
The effects of dietary replacement of conventional
inorganic zinc sources (Zn sulphate and Zn oxide)
with ZnMet on performance indices are presented in
Table 2. As shown, dietary zinc oxide supplementation caused an increase (p < 0.01) in feed consumption during the starter period. Although partial
substitution of inorganic Zn by ZnMet had no
significant effect on feed intake, however, complete
replacement of ZnMet with inorganic sources tended
(p > 0.05) to increase feed intake by the birds during
the starter and grower periods and increased
(p < 0.05) it during entire (142 days) experimental
period. The partial substitution of inorganic Zn by
ZnMet improved (p < 0.01) body weight gain during
the starting period. In addition, weight gain was
affected by replacement level during the grower and
entire experimental periods. Similarly, dietary inclusion of ZnMet partially in place of inorganic sources
improved feed conversion efficiency during both 121
(p < 0.01) and 142 (p < 0.05) days of age.
As noted in Table 3, the relative pancreas weight
was not affected by dietary treatments. On the other
hand, dietary supplementation with 100% ZnMet
increased (p < 0.05) the relative weight of liver. Dietary inclusion of ZnMet in place of inorganic Zn
sources resulted in a decrease (p < 0.01) in abdominal
fat percentage. In addition, complete substitution of
ZnMet for inorganic Zn improved (p < 0.05) carcass
yield. Interestingly, single degree of freedom contrast
comparisons showed that dietary supplementation
with Zn oxide increased both breast and carcass yields
relative to another inorganic zinc source.
The effect of dietary replacement of inorganic zinc
sources with ZnMet complex on zinc concentrations
in plasma, liver and thymus of 42-days-aged broiler
chicks is shown in Table 4. Although plasma Zn concentration was not influenced by dietary treatments
or replacement level, experimental diets had considerable impact (p < 0.05) on Zn accumulations in liver
and thymus so that, partial substitution of inorganic
Zn by ZnMet resulted in increases in Zn deposition in
both tissues.
The partial replacement of inorganic Zn sources by
ZnMet caused a reduction (p < 0.01) in plasma uric

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

Daily weight gain (g/bird)

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

Feed conversion ratio (g feed: g gain)

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

Daily feed intake (g/bird)

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

Effect of zinc-methionine on broiler performance

R. Jahanian and E. Rasouli

Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH

Effect of zinc-methionine on broiler performance

R. Jahanian and E. Rasouli

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.

complexing between Zn and amino acid contained in

ZnMet does not allow fibre or phytate to chelate Zn to
way that reduce its availability for host animal.
As pointed out in Table 2, substitution of ZnMet for
conventional inorganic Zn sources improved feed conversion ratio (FCR) during 121 and 142 days of age.
Diets supplemented with zinc-amino acid complexes
have been shown to improve FCR in broilers. Hess
et al. (2001) supplemented practical broiler diets with
40 mg/kg zinc from three different sources. Feed conversion efficiency was improved from 1 to 35 and
from 1 to 42 days of age when supplemental zincamino acid complexes were provided to female broilers (Hess et al., 2001). The better FCR values in initial
weeks by organic zinc supplementation in study here
may be attributed to proper feathering as concluded
by Jahanian et al. (2008a). Zinc acts directly in the
process of feather growth; thus, Zn deficiencies can
seriously affect feathering (Scott et al., 1982). Higher
bioavailability of organic zinc sources may supply zinc
and essential amino acids (such as Met) requirements
for rapid feathering and in turn, decrease mainte-

nance requirements because of the insulating action

of feathers. However, in a previous study, we failed to
find beneficial impacts of organic zinc supplementation on FCR, neither in starter nor in entire trial
period (Jahanian et al., 2008b).
Dietary inclusion of ZnMet in place of inorganic Zn
reduced abdominal fat percentage and improved both
breast and carcass yields. Interestingly, breast and carcass yields were observed to be higher in oxide-supplemented broilers than those on sulphate diets,
probably due to the higher feed intake in the former
group. Partial substitution of inorganic Zn by ZnMet
improved numerically (p > 0.05) carcass and breast
yields. In addition, carcass and liver of 100% ZnMetsupplemented birds were heavier in relative weights
compared with 100% sulphate-fed groups. The exact
mechanism(s) responsible for these events remain to
be elucidated; however, it seems that the higher feed
consumption and subsequently the higher growth
rates in oxide- and ZnMet-supplemented groups are
largely responsible for these observations. Of course,
the heavier livers and greater carcass yields in chicks

Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH

55

DOI: 10.1111/jpn.12213

ORIGINAL ARTICLE

Effects of dietary substitution of zinc-methionine for inorganic

zinc sources on growth performance, tissue zinc accumulation
and some blood parameters in broiler chicks
R. Jahanian and E. Rasouli
Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, Iran

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

many tissues, are most often associated with proteins

(McCall et al., 2000).
Zinc deficiency arrests cell proliferation, while in
multicellular organisms, it also results in abnormal differentiation and development, leading to extensive
abnormalities or syndromes (Christianson, 1991;
Coleman, 1992). Supplemental zinc is added to poultry diets to prevent the problems associated with Zn
deficiencies. In this case, National Research Council
(1994) recommends the level of 40 mg added zinc/kg
feed for broilers; however, the poultry rations are
often formulated to contain dietary zinc concentrations over 80 mg/kg. The higher concentrations of

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Effect of zinc-methionine on broiler performance

R. Jahanian and E. Rasouli

rapidly to the changes in Zn intake and alterations

in dietary utilizable (bioavailable) zinc content
(Buckley, 2000).
Hempe and Cousins (1989) suggested that ZnMet
and copper-lysine complexes are transported intact
from the intestinal lumen into the mucosal cells. If
ZnMet is absorbed and transported without any modification, the ZnMet complex may provide a mean for
increasing tissue supply of Zn as well as Met, which
should improve animal productivity when Met is the
first limiting amino acid (Puchala et al., 1999). On the
other hand, reports show that Zn has a critical and
important role in protein turnover throughout the
body and affect protein degradation and its metabolic
wastes (Christianson, 1991; Kaim and Schwederski,
1994). This is a reasonable explanation why plasma
References
Ammerman, C. B.; Baker, D. H.; Lewis, A.
J., 1995: Bioavailability of Nutrients for
Animals: Amino Acids, Minerals and Vitamins. Academic Press, San Diego, CA.
Anonymous, 1982: Analytical Methods for
Atomic Absorption Spectrophotometry. Perkin-Elmer Corp, Nor Walk, CT.
Association of Official Analytical Chemists, 1995: Official Methods of Analysis,
16th edn. AOAC, Washington, DC.
Baker, D. H.; Ammerman, C. B., 1997:
Zinc bioavailability. In: C. B. Ammerman, D. H. Baker, A. J. Lewis (eds), Bioavailability of Nutrients for Animals: Amino
Acids, Minerals, and Vitamins. Academic
Press, San Diego, CA, pp. 367398.
Baker, D. H.; Halpin, K. M., 1988: Zinc
antagonizing effect of fish meal, wheat
bran and a corn soybean meal mixture
when added to a phytate- and fiber-free
casein dextrose diet. Nutrition Research 8,
213.
Batal, A. B.; Parr, T. M.; Baker, D. H.,
2001: Zinc bioavailability in tetrabasic
zinc chloride and the dietary zinc
requirement of young chicks fed a soy
concentrate diet. Poultry Science 80,
8790.
Buckley, W. T., 2000: Trace element
dynamics: zinc. In: J. P. E. DMello
(ed.), Farm Animal Metabolism and Nutrition. CABI Publishing, Wallingford, UK,
pp. 163173.
Burrell, A. L.; Dozier, W. A. III; Davis, J.
A.; Compton, M. M.; Freeman, M. E.;
Vendrell, P. F.; Ward, T. L., 2004:
Responses of broilers to dietary zinc
concentrations and sources in relation

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.

to environmental implications. British

Poultry Science 45, 255263.
Cao, J.; Henry, P. R.; Davis, S. R.; Cousins,
R. J.; Miles, R. D.; Littell, R. C.;
Ammerman, C. B., 2002: Relative
bioavailability of organic zinc sources
based on tissue zinc and metallothionein in chicks fed conventional dietary
zinc concentrations. Animal Feed Science
and Technology 101, 161170.
Cheng, J.; Kornegay, E. T.; Schell, T. C.,
1998: Influence of dietary lysine on the
utilization of zinc from zinc sulfate and
a zinc-lysine complex by young pigs.
Journal of Animal Science 76, 10641074.
Christianson, D. W., 1991: The structural
biology of zinc. Advances in Protein Chemistry 42, 281335.
Coleman, J. E., 1992: Zinc proteins:
enzymes, storage proteins, transcription
factors, and replication proteins. In: C.
C. Richardson, J. N. Abelson, A. Meister,
C. T. Walsch (eds), Annual Review of Biochemistry. Annual Reviews, Palo Alto,
CA, pp. 897946.
Davies, N. T.; Hristic, V.; Flett, A. A., 1977:
Phytate rather than fiber in bran as the
major determinant of zinc availability to
rats. Nutrition Reports International 15,
207.
Edwards, H. M. Jr; Baker, D. H., 1999:
Bioavailability of zinc in several sources
of zinc oxide, zinc sulfate, and zinc
metal. Journal of Animal Science 77,
27302735.
Ellis, R.; Morris, E. R.; Hill, A. D., 1982:
Bioavailability to rats of iron and zinc in
calcium-iron-phytate and calcium-zincphytate complex. Nutrition Research 2,
319322.

Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH

Fordyce, E. J.; Forbes, R. M.; Robbins, K.

R.; Erdman, J. W. Jr, 1987: Phytate9calcium/ zinc molar ratios: are they predictive of zinc bioavailability? Journal of
Food Science 52, 421428.
Hahn, J. D.; Baker, D. H., 1993: Growth
and plasma zinc responses of young pigs
fed pharmacologic levels of zinc. Journal
of Animal Science 71, 30203024.
Hempe, J. M.; Cousins, R. J., 1989:
Effect of EDTA and zinc-methionine
complex on zinc absorption by rat
intestine. The Journal of Nutrition 119,
11791187.
Hess, J. B.; Bilgili, S. F.; Parsons, A. M.;
Downs, K. M., 2001: Influence of complexed zinc products on live performance and carcass grade of broilers.
Journal of Applied Poultry Research 19,
4960.
Jahanian, R.; Yaghoubi, M. J., 2010: Dietary zinc source and level affect cellular
and humoral immune functions in broiler chickens. In: 2nd International Veterinary Poultry Congress. 2021 February,
Tehran, Iran. pp. 201.
Jahanian, R.; Nassiri Moghaddam, H.; Rezaei, A., 2008a: Improved broiler chick
performance by dietary supplementation of organic zinc sources. AsianAustralian Journal of Animal Sciences 21,
13481354.
Jahanian, R.; Nassiri Moghaddam, H.; Rezaei, A.; Haghparast, A. R., 2008b: The
influence of dietary zinc-methionine
substitution for zinc sulfate on broiler
chick performance. Journal of Biological
Sciences 8, 321327.
Kaim, W.; Schwederski, B., 1994: Bioinorganic Chemistry: Inorganic Elements in the

57

Effect of zinc-methionine on broiler performance

Chemistry of Life. John Wiley and Sons,

Chichester, UK, pp. 401.
Keilin, D.; Mann, T., 1940: Carbonic
anhydrase. Purification of the
enzyme. Biochemical Journal 34,
11631176.
Lesson, S.; Summers, J. D., 2001: Scotts
Nutrition of the Chickens, 4th edn. Uni
Book, Guelph, ON, Canada.
McCall, K. A.; Huang, C.; Fierke, C. A.,
2000: Function and mechanism of zinc
metalloenzymes. The Journal of Nutrition
130, 1437S1446S.
Nassiri Moghaddam, H.; Jahanian, R.,
2009: Immunological responses of broiler chicks can be modulated by dietary
supplementation of zinc-methionine in
place of inorganic zinc sources. AsianAustralian Journal of Animal Sciences 22,
396403.
National Research Council, 1994: Nutrient
Requirements of Poultry, 9th rev. edn.
National Academy Press, Washington,
DC.
Puchala, R.; Sahlu, T.; Davis, J. J., 1999:
Effects of zinc-methionine on perfor-

58

R. Jahanian and E. Rasouli

mance of Angora goats. Small Ruminant

Research 33, 18.
Sandoval, M.; Henry, P. R.; Ammerman,
C. B.; Miles, R. D.; Littell, R. C., 1997a:
Relative bioavailability of supplemental
inorganic zinc sources for chicks. Journal
of Animal Science 75, 31953205.
Sandoval, M.; Henry, P. R.; Littell, R. C.;
Cousins, R. J.; Ammerman, C. B.,
1997b: Estimation of the relative bioavailability of zinc from inorganic zinc
sources for sheep. Animal Feed Science
and Technology 66, 223235.
SAS Institute, 1999: SAS Statistics Users
Guide. Statistical Analytical System, 5th
rev. edn. SAS Institute Inc, Carry, NC.
Schell, T. C.; Kornegay, E. T., 1996:
Zinc concentration in tissues and performance of weanling pigs fed pharmacological levels of zinc from ZnO,
Zn-methionine, Zn-lysine, or ZnSO4.
Journal of Animal Science 74, 1584
1593.
Scott, M. L.; Nesheim, M. C.; Young, R. J.,
1982: Nutrition of the Chicken, 3rd edn.
W. F. Humphrey Press, Geneva, NY.

Spears, J. W., 1989: Zinc methionine

for ruminants: relative bioavailability
of zinc in lambs and effects of
growth and performance of growing
heifers. Journal of Animal Science 67,
835843.
Steel, R. G. D.; Torrie, J. H.; Dickey, D. A.,
1997: Principles and Procedures of Statistics:
A Biometrical Approach, 3rd edn.
McGraw-Hill, New York, USA.
Vallee, B. L.; Auld, D. S., 1990: Active-site
zinc ligands and activated H2O of zinc
enzymes. Proceedings of the National
Academy of Sciences of the United States of
America 87, 220224.
Wedekind, K. J.; Hortin, A. E.; Baker, D.
H., 1992: Methodology for assessing zinc
bioavailability: efficacy estimates for zincmethionine, zinc sulfate, and zinc oxide.
Journal of Animal Science 70, 178187.
Wedekind, K. J.; Lewis, A. J.; Giesemann,
M. A.; Miller, P. S., 1994: Bioavailability
of zinc from inorganic and organic
sources for pigs fed corn-soybean meal
diets. Journal of Animal Science 72,
26812689.

Journal of Animal Physiology and Animal Nutrition 2014 Blackwell Verlag GmbH