The effects of in ovo feeding of glutamine in broiler breeder eggs on hatchability, development of the gastrointestinal tract, growth performance and carcass characteristics of broiler chickens

Mehdi Salmanzadeh , Yahya Ebrahimnezhad , Habib Aghdam Shahryar , and Jamshid Ghiasi Ghaleh-Kandi

Departments of Animal Science , Shabestar branch , Islamic Azad University, Shabestar, Iran

Correspondence to: Yahya Ebrahimnezhad (ebrahimnezhad@gmail.com)

Received: 27 February 2016 – Revised: 30 April 2016 – Accepted: 17 May 2016 – Published: 31 May 2016

Abstract: The aim of the present study was to investigate the effect of in ovo feeding (IOF) of glutamine on hatchability, development of the gastrointestinal tract, growth performance and carcass characteristics of broiler chickens. Fertilized eggs were subjected to injections with glutamine (Gln) (10, 20, 30, 40 or 50 mg dissolved in 0.5 mL of dionized water) on day 7 of incubation. Hatchability, growth performance, carcass characteristics (carcass weight and relative weights of breast, thigh, heart, liver, gizzard, abdominal fat, intestine, pancreas and spleen) and jejunal morphometry (measurement of villus height and width and crypt depth) were determined during the experiment. The weight of newly hatched chickens was significantly greater in groups with Gln injection than in control and sham groups. But IOF caused lower hatchability than in the control group (non- injected eggs) (p < 0.05). Chickens from IOF of Gln showed better weight gain and feed conversion ratio (0– 42 days of age), when compared to chickens hatched from control and sham groups. The IOF of Gln significantly increased villus height, villus width and crypt depth at hatch period and villus height at 42 days of age. In addition, carcass weights and relative weights of breast, thigh and gizzard were also markedly increased in chickens treated in ovo with Gln; whereas heart, liver, abdominal fat, intestine, pancreas and spleen were not significantly altered at the end of the experimental period. These data suggest that the IOF of Gln may improve jejunum development, leading to an increased nutrient assimilation and consequently to greater performance in broiler chickens.

1 Introduction

In ovo feeding (IOF) is a method of supplementing exoge- nous nutrients into amnion of the avian embryo (Uni and Ferket, 2003), which can improve the performance of chicks from injected eggs (Salmanzadeh, 2011; Salmanzadeh et al., 2012; Dong et al., 2013). Previous studies have been con- ducted to assess nutrients for in ovo injection in broiler em- bryos, such as carbohydrates (Tako et al., 2004a; Uni et al., 2005; Smirnov et al., 2006), amino acids (Al-Murrani, 1982; Ohta et al., 1999, 2001, 2004), vitamins (Nowaczewski et al., 2012) and minerals (Tako et al., 2004b). However, there are few studies on IOF of Gln in broiler breeder hen eggs.

Glutamine (Gln) is the principle metabolic fuel for devel- opment of the gastrointestinal tract (Andrew and Griffiths, 2002), and is considered as a non-essential amino acid since most animal cells can synthesize it, which can play an im- portant role in the synthesis of many biologically impor- tant molecules (Souba, 1993). These results are in agree- ment with the findings of Samli et al. (2007), who reported that Gln is an important amino acid for utilization as an en- ergy source for the development of the gastrointestinal tract and stimulates intestinal cell proliferation, which leads to in- creasing the absorptive source of the gastrointestinal mucosa and consequently the access to nutrients. In parallel, previ- ous experiments, showed that supplementing the diet with

236 M. Salmanzadeh et al.: The effects of in ovo feeding of glutamine in broiler breeder eggs

Item

Diet

Starter 0–10 days

Grower 11–26 days

Finisher 27–42 days

Ingredient (%)

Corn

60.36

65.44

66.8

Soybean meal (44 % CP)

34.12

28.62

26.33

Vegetable fat

1.23

1.74

2.84

Dicalcium phosphate

1.83

1.8

1.67

Oyster sell-ground

1.22

1.19

1.13

Salt

0.35

0.3

0.3

Sodium bicarbonate

0.11

0.07

0.07

Vitamin premix1

0.25

0.25

0.25

Trace mineral premix2

0.25

0.25

0.25

DL-Met

0.17

0.18

0.18

L-Lys

0.11

0.16

0.18

Calculated analysis

ME (kcal kg−1)

2894

2987

3176

CP (%)

20.3

18.3

18

Ca (%)

1

0.96

0.9

Available P (%)

0.50

0.48

0.45

Met (%)

0.46

0.44

0.43

Met + cystine (%)

0.89

1.20

0.84

1.10

0.82

1.05

Table 1. Composition and calculated contents of nutrients of broiler diet.

Lys (%)

1 Vitamin premix provided the following per kilogram of diet: vitamin A, 11 013 IU; vitamin D3, 3525 IU; vitamin E, 33 IU; vitamin K, 2.75 mg; riboflavin, 7.7 mg; pantothenic acid, 17.6 mg; niacin, 55.1 mg; choline, 478 mg; vitamin B12,

0.028 mg; pyridoxine, 5.0 mg; thiamine, 2.2 mg; folic acid, 1.1 mg; biotin, 0.22 mg. 2 Trace mineral premix provided the following per kilogram of diet: manganese, 64 mg; zinc, 75 mg; iron, 40 mg; copper, 10 mg; iodine, 1.85 mg; and selenium,

0.3 mg.

Gln increased intestinal villus height and consequently, im- proved growth performance in broilers (Bartell and Batal, 2007; Yi et al., 2005; Jazideh et al., 2014), turkey poults (Yi et al., 2001; Salmanzadeh and Shahryar, 2013a) quails (Salmanzadeh and Shahryar, 2013b) and weanling pigs (Kitt et al., 2002), compared to control groups. Chen et al. (2009) demonstrated that, body weights measured at the 7-days old post-hatching were significantly modified in ducks treated in ovo injection with Gln.

All these beneficial actions of Gln in particular, make it an amino acid deserving of scientific and technical attention.

It was hypothesized that IOF of Gln can improve the de- velopment of the gastrointestinal tract, by increasing the ab- sorptive surface area that consequently promotes the nutri- ent assimilation and improves growth performance in broiler chickens. Thus, the aims of this research were to investigate the effects of IOF of Gln in broiler breeder eggs in hatchabil- ity, development of the gastrointestinal tract, growth perfor- mance and carcass characteristics of broiler chickens.

  • Material and method

  • Incubation and in ovo feeding

±

Hatching eggs of approximately similar weights (65 1 g) were obtained from broiler breeder strain (Cobb 500) at 42 weeks of age. A total of 1400 fertile eggs were numbered, fumigated with formaldehyde gas, and incubated at 37.7 C and 64 % RH. On the 6th day of incubation, the eggs were

candled, and the infertile ones or those containing only dead embryos were removed. On the 7th day, fertile eggs were randomly allotted to seven treatments with four replicate per treatment and 50 eggs per replicate. Eggs were injected with Gln (10, 20, 30, 40, and 50 mg, respectively) dissolved in

0.5 mL of deionized water whereas in the sham group, eggs were only treated with deionized water (0.5 mL) and those of the control group received no injection.

Before injection, each egg was candled to identify the lo- cation of the future injection. The injection hold area was cleaned with ethyl alcohol (70 %) and the blunt end was punched using a 22 gauge needle. Then, the solutions were

injected into albumen (0.5 mL egg−1) using another dispos- able syringe equipped with a 22 gauge needle to a depth of

about 13 mm (Salmanzadeh et al., 2012). After injection, the

  1. Salmanzadeh et al.: The effects of in ovo feeding of glutamine in broiler breeder eggs 237

eggs were sealed with cellophane tape, and transferred to the incubator. Control eggs were removed from the incubator to- gether with the treated groups, and kept in the same environ- ment. The group of eggs designated as sham-injected con- trols were injected with 0.5 mL of deionized water. Deion- ized water injections were included as sham controls primar- ily to rule out a possible negative response caused by the stress of injection and handling. L-Glutamine was supplied from Sigma® Co (anhydrous 99 %, CAS Number: 56-85- 9). All of the treatment solutions were prepared in autoclaved water.

  • Birds and data collection

After hatching, chickens were transferred to an experimental house and reared for 42 days with the same ration according to the requirements of broiler as recommended by the catalog of Cobb 500 broilers (Table 1). Each chicken according to the treatment group was identified by the neck tag and recorded.

Table 2. Effects of IOF of Gln on weight and hatchability in newly hatched chickens.

Groups Hatchability (%) Weight (g) Control 89.58a 43.41e

Sham group 72.22c 43.31e

Gln1 10 mg 75.69bc 43.82cd

Gln 20 mg 75.00bc 43.71d

Gln 30 mg 76.38bc 44.09ab

Gln 40 mg 79.16b 43.93bc

Gln 50 mg 77.77bc 44.21a SEM 1.75 0.05

P Value 0.0001 0.0001

=

1 Gln Glutamine; a–e averages in a column with different superscript letters are significantly different.

Table 3. Effects of IOF of Gln on body weight gain (BWG), food intake (FI) and feed conversion ratio (FCR) of broilers to 1–10 days of age.

All chickens and treatments were randomly assigned to 1 of

28 pens. Each open was provided with water and an individ- ual feeder. Room temperature was maintained at 32 C from

Groups 1–10 days of age (g)

BWG FI FCR

0 to 4 and then gradually reduced from 32 to 21 C. All ex-

perimental protocols and procedures were approved by the institutional Animal Care of Iran.

Upon hatching, the number of hatchlings was determined to calculate the hatchability of fertile eggs. The weight of newly hatched chickens was determined by weighing all chickens hatched one by one. In each pen, bird body weight and food intake were recorded on days 1, 10, 26, and 42 post- hatching and thereafter mean body weight gain, food intake, and food conversion ratio were calculated for each pen (repli- cate) between 0 and 10, 11 and 26, 27 and 42, and 1 and 42 days. In each time period, body weight gain was calcu- lated and expressed as grams per bird. Food intake (g of food intake/bird) over the entire grow-out period was calculated by totalling food consumption in each time interval between each bird sampling. Food conversion ratio (g of food intake/g of body weight gain) was calculated by dividing total food intake by total weight gain in each pen.

  • Morphometric indices of the jejunum

At hatching and 10 days of age, eight birds from pre- treatment were euthanized by cervical dislocation. Then, gas- trointestinal tract was carefully excised. One cross-section for jejunum was fixed with formalin solution and was pre- pared using standard paraffin embedding procedures by sec- tioning at 5 mm thickness, and staining with hematoxylin and eosin. Villus height (VH), villus width (VW) and crypt depth (CD) were determined using an image processing and ana- lyzing system (Image Pro plus) and were expressed as mi- crometers (Touchette et al., 2002).

Control 199.09bc 251.25 1.26ab

Sham group

196.03c

255.78

1.30a

Gln 10 mg

206.21ab

251.98

1.22abc

Gln 20 mg

207.87a

245.00

1.17bc

Gln 30 mg

209.98a

246.97

1.17bc

Gln 40 mg

206.49ab

250.14

1.21abc

Gln 50 mg

213.70a

243.24

1.14c

SEM

2.53

6.49

0.03

P Value

0.0010

0.8358

0.0237

a–c Averages in a column with different superscript letters are significantly different.

  • Carcass measurements

On day 42, two broilers per pen were selected (close to the mean weight of each cage), weighed and killed by cervi- cal dislocation, and then the abdominal cavity was opened. Weights of the eviscerated hot carcass, breast muscle, thigh, liver, heart, gizzard, abdominal fat, intestine, pancreas and spleen were recorded and the corresponding percentages (% of live body weight) were calculated.

  • Statistical analysis

±

Analyses of variance were performed using the GLM proce- dure of SAS Institute Inc. (2005) as a completely random- ized design. Results are presented as mean SEM. The sig- nificantly different treatment means were investigated using Duncan’s new multiple rang test. Differences were consid- ered significant when p < 0/05.

238 M. Salmanzadeh et al.: The effects of in ovo feeding of glutamine in broiler breeder eggs

Table 4. Effects of IOF of Gln on body weight gain (BWG), food intake (FI) and feed conversion ratio (FCR) of broilers to 11– 26 days of age.

Table 6. Effects of IOF of Gln on body weight gain (BWG), food intake (FI) and feed conversion ratio (FCR) of broilers to 1–42 days of age.

Groups 11–26 days of age (g) Groups 1–42 days of age (g)

BWG

FI

FCR

BWG

FI

FCR

Control

647.74bc

915.24

1.41a

Control

2250.26c

4318.99

1.91a

Sham group

639.75c

917.80

1.43a

Sham group

2233.74c

4317.08

1.93a

Gln 10 mg

673.12ab

909.36

1.35bc

Gln 10 mg

2331.91ab

4300.86

1.84b

Gln 20 mg

669.22abc

907.66

1.35b

Gln 20 mg

2325.34b

4287.69

1.84b

Gln 30 mg

686.50a

904.98

1.31bc

Gln 30 mg

2352.08ab

4276.20

1.81bc

Gln 40 mg

679.96a

892.87

1.31bc

Gln 40 mg

2359.43ab

4258.06

1.80bc

Gln 50 mg

692.19a

895.13

1.29c

Gln 50 mg

2389.61a

4263.36

1.78c

SEM

9.68

8.23

0.01

SEM

19.69

22.60

0.01

P Value

0.0074

0.2981

0.0001

P Value

0.0001

0.3558

0.0001

a–c Averages in a column with different superscript letters are significantly different.

Table 5. Effects of IOF of Gln on body weight gain (BWG), food intake (FI) and feed conversion ratio (FCR) of broilers to 27– 42 days of age.

Groups 27–42 days of age (g) BWG FI FCR

Control 1403.42b 3152.50 2.24a

Sham group 1397.97b 3143.50 2.24a

Gln 10 mg

1452.57a

3139.52

2.16b

Gln 20 mg

1448.25a

3135.03

2.16b

Gln 30 mg

1455.59a

3124.25

2.14bc

Gln 40 mg

1472.97a

3115.05

2.11bc

Gln 50 mg

1483.72a

3124.99

2.10c

SEM

11.41

11.54

0.01

P Value

0.0001

0.3229

0.0001

a–c Averages in a column with different superscript letters are significantly different.

  • Results

The effects of IOF with different levels of Gln solutions at 7 days of incubation on hatchability and weights of newly hatched chickens are presented in Table 2. IOF with Gln and deionized water reduced the hatchability compared with the control group (not injection eggs). But, the mean body weights of newly hatched chickens injected by Gln were increased more than the control groups. IOF of Gln im- proved body weight gain and feed conversion ratio through the whole experimental period but feed intake was not sig- nificantly altered (Tables 3–6).

As seen in Tables 7 and 8, villus height, villus width and crypt depth in jejunum were significantly increased in chick- ens treated in ovo by Gln compared to the non-injected and sham controls in newly hatched and 10-day old chickens.

a–c Averages in a column with different superscript letters are significantly different.

Control

342.24ef

49bc

50.50d

Sham group

338.19f

48c

49.25d

Gln 10 mg

357.89bcd

55ab

60.00bc

Table 7. Effects of IOF of Gln on villus height, villus width and crypt depth in jejunum of newly hatched chickens.

Groups Villus Villus Crypt height (µm) width (µm) depth (µm)

Gln 20 mg

355.48bcde

58a

59.00c

Gln 30 mg

352.25cde

56a

65.25abc

Gln 40 mg

360.49ab

59a

68.00ab

Gln 50 mg

372.07a

60a

70.00a

SEM

4.37

2.13

2.71

P Value

0.0001

0.0012

0.0001

a–e Averages in a column with different superscript letters are significantly different.

On the 42nd day, weights of the carcass, breast, thigh and gizzard were also significantly increased in broilers from in- jected eggs with Gln compared to the control groups (p < 0.05); the highest value was observed in the group receiv- ing 50 mg Gln. By contrast, the liver, heart, abdominal fat, intestine, pancreas and spleen weights were not significantly altered in broilers from Gln treated eggs (Tables 9, 10).

  • Discussion

Concerning hatchability, Uni et al. (2005) showed that the positive effects of IOF at late-term chicken embryos may in- clude increased hatchability. On the other hand, Pedroso et al. (2006a) reported that IOF of Gln in the amniotic fluid of embryos on day 18 of incubation had no significant effect on the hatchability. In parallel, Dos Santos et al. (2010) demon- strate that, IOF of 0.5 mL of a 10 % Gln solution did not af- fect hatching of newly hatched chickens. In contrast, Ohta

  1. Salmanzadeh et al.: The effects of in ovo feeding of glutamine in broiler breeder eggs 239

Table 8. Effects of IOF of Gln on villus height, villus width and crypt depth in jejunum of broilers when they were 10-days old.

Table 9. Effects of IOF of Gln on weight of carcass, breast and thigh of broiler chickens when they were 42-days old (based on percentage of live body weight).

Groups Villus Villus Crypt

height (µm)

width (µm)

depth (µm)

Groups

Carcass (%)

Breast (%)

Thigh (%)

Control

656.25c

139.00d

86b

Control

67.56d

23.92c

19.96c

Sham group

647.50c

141.75cd

85b

Sham group

67.71d

24.21c

19.82c

Gln 10 mg

677.00b

152.75abc

94a

Gln 10 mg

68.58c

24.94b

20.68b

Gln 20 mg

687.50ab

162.50a

98a

Gln 20 mg

68.48c

24.89b

20.64b

Gln 30 mg

684.25ab

151.75abc

97a

Gln 30 mg

68.71bc

25.55a

21.35a

Gln 40 mg

691.00ab

155.75ab

96a

Gln 40 mg

68.97ab

25.78a

21.48a

Gln 50 mg

702.75a

158.50a

95a

Gln 50 mg

69.18a

25.86a

21.61a

SEM

6.99

3.70

1.87

SEM

0.08

0.11

0.09

P Value

0.0001

0.0009

0.0001

P Value

0.0001

0.0001

0.0001

a–e Averages in a column with different superscript letters are significantly different.

+

et al. (1999) showed that the hatchability was significantly reduced when injecting amino acids at day 0 of incubation. Chen et al. (2010) stated that hatchability of control, sources and maltose (DS), L-alanyl–L-glutamine (Ala–Gln), Su- crose, maltose and L-alanyl–L-glutamine (DS Ala–Gln), groups were 85, 65, 70 and 82 % respectively. Moreover, DS and Ala–Gln decreased the hatchability by 24 and 18 % (p < 0.01), respectively. In an experiment by Chen et al. (2010), the pH of all the injection solution was adjusted to about

+

6.8 and the osmolarity was 154, 2264 and 30 mOsm for DS, Ala–Gln and DS Ala–Gln solution respectively. These re- searchers showed that decreasing in hatchability was due to varied osmolarity of the injection. Previous work demon- strated that the osmolarity of injection solution was a criti- cal factor in affecting the hatchability of the chick (Ferket et al., 2005). In parallel, Pedroso et al. (2006b) also found that decreased hatchability was observed when chick embryos re- ceived glucose in ovo injection at 16 days of incubation. The significant difference in hatchability among the in ovo in- jected groups indicated that nutrient specificity might result in the differing response of embryos. According to the US Patent (6592878) of Uni and Ferket (2003) the importance of the osmolarity of the in ovo feeding solution on hatch- ability of chicks was illustrated. Acceptable hatchability of chicks was observed when eggs were injected with solutions having an osmolarity ranging below 800 mOsm with an opti- mum hatchability observed at about 400–600 mOsm. Unac- ceptable hatching rates were observed when the in ovo feed- ing solution exceeded 800 mOsm. In the present study osmo- lality of all the injection of solutions was adjusted to about 450 mOsm. Thus, osmolarity could not be the reason for the decreasing of the hatchability. This is consistent with the re- sults obtained in the present study, in which the injection of different levels of Gln and deionized water caused significant decrease in the hatchability. Probably the reducing hatchabil- ity was due to the IOF into the albumin. One of the possible reasons for the decreasing rate of hatching is the allergic cav-

a–d Averages in a column with different superscript letters are significantly different.

ity under the air sac causing the respiration of developing embryo to stop and die. Heiblum et al. (2001) showed that IOF of glucocorticoid at day 7 of incubation resulted in 35 % decline of rate of hatching. Also, Salmanzadeh et al. (2012) observed lower hatchability when 7-day old broiler embryos were injected with glucose and magnesium. Thus, it seems that any IOF at early embryonic life can be harmful for the internal environment susceptibility and have negative effects on rate of hatching. This effect may largely independent de- pend on injected Gln effect.

Data of this study showed that the IOF of Gln into the albu- min can be seen as an effective tool to improve the mean body weights of newly hatched chickens. Previous studies showed that to stimulate the development of the chick embryo is an important factor in increasing the weight of newly hatched chickens (Tako et al., 2004a; Uni and Ferket, 2004; Uni et al., 2005; Smirnov et al., 2006). Al-Murrani (1982) showed that IOF of amino acid into fertile goose eggs at day 7 of incubation increased the weight of newly hatched geese. In turkeys, Foye et al. (2006) observed that in ovo injection of egg white protein at day 23 of incubation increased hatch- ing weight. Ohta et al. (2001) reported that IOF of amino acids into the air cell of 7-day old embryos improved uti- lization of amino acid by the embryo and consequently in- creased hatching weight. On the other hand, Dos Santos et al. (2010) showed that IOF with 10 % Gln at 18 days of in- cubation did not influence hatch weight. In another study, Chen et al. (2009) showed that in ovo administration of Gln and carbohydrates in combination in the duck did not affect the body weight at 25 days of incubation, weight of hatch and weight of ducks on day 3 post-hatching. According to the past studies and our present observations, the late-term embryo and neonatal chicken depends on gluconeogenesis from amino acids, resulting in the depletion of muscle pro- tein reserves and the reduction of hatching weight. To re- duce the depletion of muscle protein, we carried out IOF Gln into albumen prior to hatching, which would support the en-

240 M. Salmanzadeh et al.: The effects of in ovo feeding of glutamine in broiler breeder eggs

Table 10. Effects of IOF of Gln on weight of heart, liver, gizzard, abdominal fat, intestine, pancreas and spleen of broiler chickens when they were 42-days old (based on percentage of live body weight).

Groups

Heart (%)

Liver (%)

Gizzard (%)

Abdominal fat (%)

Intestine(%)

Pancreas (%)

Spleen (%)

Control

0.58

2.09

1.70b

2.13

4.49

0.143

0.126

Sham group

0.60

2.11

1.69b

2.16

4.45

0.148

0.121

Gln 10 mg

0.64

2.13

2.01a

2.23

4.68

0.158

0.129

Gln 20 mg

0.64

2.18

1.99a

2.34

4.62

0.156

0.133

Gln 30 mg

0.68

2.24

1.96a

2.32

4.85

0.160

0.135

Gln 40 mg

0.69

2.18

2.09a

2.36

4.92

0.164

0.141

Gln 50 mg

0.73

2.21

2.11a

2.42

5.04

0.170

0.142

SEM

0.06

0.09

0.05

0.08

0.16

0.009

0.004

P Value

0.7562

0.9168

0.0001

0.1251

0.2058

0.6085

0.4787

a–b Averages in a column with different superscript letters are significantly different.

ergy status of the hatching by moderating the use of muscle that consequently increases body weight at hatching. In ad- dition, Foye et al. (2006) showed that the supply of nutrient by IOF improved avian energy status, which spared energy used for metabolism and, consequently, increased postnatal performance. Today, whole embryonic life is almost 35 % of the productive life of broilers. Thus, to stimulate the develop- ment of the chick embryo is an important factor to increase the weight of newly hatched chicks. Also, previous stud- ies showed that weight of newly hatched chickens is a ma- jor predictor for marketing weight in modern broilers. Wil- son (1991) stated that each 1g of improvement in weight of newly hatched chickens leads to 8–13 g of improvement in body weight at marketing. In this study we demonstrate that a 1 g difference in weight of newly hatched chickens due to IOF resulted in 109–130 g increase in body weight at day 42. Based on the results of this study, the growth perfor- mance of broiler chickens linearly increased with the im- proved development of the gastrointestinal tract. Thus, im- proved growth performance was dependent upon the devel- opment of the gastrointestinal tract and IOF of Gln used. Pre- vious studies showed that the dietary supplementation with Gln stimulates the intestine development and improved the growth performance of turkey poults first week after hatch (Yi et al., 2001), weanling pigs (Kitt et al., 2002), broilers (Bartell and Batal, 2007) and Japanese quails (Salmanzadeh

and Shahryar, 2013b).

It is well demonstrated that Gln appears to be a condi- tionally essential amino acid nutrient as exogenous Gln can be a potential candidate in improving intestinal morphology and digestive function (Alverdy et al., 1992; Helton et al., 1990; Shizuka et al., 1990). On the other hand, from the con- clusions above, it is concluded that the increase in intesti- nal villus height reported in animals fed with Gln supple- mented diets may explain the improvement in growth per- formance. Therefore, the increased villus height may am- plify nutrient absorption and utilization of nutrients leading to improved growth performance of broiler chickens. On the

contrary, Chen et al. (2009) reported that body weights mea- sured at the 25th day of incubation, at hatch (Day 0), and at the 3rd day post-hatching were not significantly modified in ducks treated in ovo with Gln, whereas IOF of Gln improved small intestine development and body weight than the control ducks when they were 7-days old.

In the present study, the relative weights of carcass, breast, thigh and gizzard were also significantly improved in broiler chickens injected with Gln. Chen et al. (2009) demonstrated that the in ovo injection of Gln at day 21 of incubation im- proved breast weight of ducks at 25 d of incubation un- til day 7 of post-hatch, whereas gizzard, proventriculus and liver weight were not significantly altered. Salmanzadeh and

Shahryar (2013b) showed that dietary Gln supplementation (0, 20, 30, 40 and 50 mg kg−1 of diet) significantly increased the relative weights of carcass and breast compared to the control quails whereas liver and gizzard weights were not significantly modified.

Dai et al. (2009) demonstrated that dietary Gln addition (5 and 10 g kg−1) improved growth performance and carcass traits of broilers. Furthermore, in another study, increases in carcass weight and relative weight of breast and thigh were also significantly improved in broilers supplemented with Gln at 5 g kg−1 of diet (Dai et al., 2011).

5 Conclusions

As a conclusion, IOF of Gln into broiler breeder eggs stimu- lated development of gastrointestinal tract and consequently, improved the growth performance and carcass characteristics whereas hatchability significantly depressed in all injected eggs compared to the not injected ones.

Author contributions. M. Salmanzadeh carried out the experi- ment and prepared the manuscript. Y. Ebrahimnezhad, H. Aghdam Shahryar, J. Ghiasi Ghaleh-Kandi designed and coordinated the ex- periment and helped to modify the manuscript.

  1. Salmanzadeh et al.: The effects of in ovo feeding of glutamine in broiler breeder eggs 241

Acknowledgements. This article is a part of PhD thesis in animal science, Islamic Azad University, Shabestar Branch (thesis supervisors: Y. Ebrahimnezhad and H. A. Shahryar, consulting advisor: J. Ghiasi Ghaleh-Kandi). Also, the authors have highly appreciated the cooperation of Saeid Ashrafi and Ayob heshmati sis for taking care of birds during the experiment period and helping in the process of experimental work.

Edited by: S. Maak

Reviewed by: two anonymous referees

References

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