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1 Abstract
  M. Fouda , A. Ateya, I. EL Araby, and A. Elzeer

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COMPUTATIONAL ANALYSIS OF PHYLOGENETIC RELATIONSHIP OF TAGAP - T CELL

2309-8694
ACTIVATION RHO GTPASE ACTIVATING PROTEIN IN SELECTED MAMMALIAN SPECIES
2 Abstract
S. H. Abbas, A. Mumtaz, M. J. ul Hasanain, M. E. Babar and M. T. Pervez

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ASSOCIATION OF POLYMORPHISMS IN PROLACTIN RECEPTOR AND MELATONIN RECEPTOR 1C

GENES ON EGG PRODUCTION AND EGG QUALITY TRAITS OF JAPANESE QUAILS (COTURNIX
COTURNIX JAPONICA)
3 Abstract
N. T. H. Nhan, L. T. T. Lan, L. T. Hung, R. L. Soubra, T. T. Gia, L. H. Anh, N. H. Xuan and N. T. Ngu

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ASSESSMENT OF DELETERIOUS EFFECTS OF BISPHENOL A ON STEROIDOGENESIS, SPERM

COUNT, AND SPERMATOGENESIS IN A MAMMALIAN MODEL
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F. Nawaz, Asmatullah and C. Ara

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LYSINE SUPPLEMENTATION IMPROVES NUTRIENTS DIGESTION, GROWTH PERFORMANCE AND

LIVER FUNCTION OF FEMALE BLUE FOXES (Alopex lagopus) IN GROWING PHASE
5 Abstract
Q. Jiang, F. Yang, X. Gao and X. Wu

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EVALUATION OF PREDICTIVE ABILITY OF TWO ARTIFICIAL NEURAL NETWORK ALGORITHMS

AND MULTIPLE REGRESSION MODEL FOR MEAT QUALITY TRAITS AFFECTED BY PRE-
SLAUGHTER FACTORS
6 Abstract
G. Ser, C. T. Bati, E. Arık and S. Karaca

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https://www.thejaps.org.pk/Volume/2021/31-06/index.php 1/5
 Journal
Lan et al.,of Animal & Plant Sciences, 31(6): 2021, Page: 1559-1567 The J. Anim. Plant 083-2021
Sci., 31 (6) 2021
ISSN (print): 1018-7081; ISSN (online): 2309-8694
https://doi.org/10.36899/JAPS.2021.6.0359
20SSOCIATION OF POLYMORPHISMS IN PROLACTIN RECEPTOR AND
MELATONIN RECEPTOR 1C GENES ON EGG PRODUCTION AND EGG QUALITY
TRAITS OF JAPANESE QUAILS (COTURNIX COTURNIX JAPONICA)
L. T. T. Lan1,#, N. T. H. Nhan2,#, L. T. Hung1, R. L. Soubra1, T. T. Gia2, L. H. Anh2, N. H. Xuan3 and N. T. Ngu2,*

1
School of Agriculture and Aquaculture, Tra Vinh University, Tra Vinh City, Vietnam
2
College of Agriculture, Can Tho University, Can Tho City, Vietnam
3
College of Food Technology and Biotechnology, Can Tho University of Technology, Can Tho City, Vietnam
*
Corresponding Author: ntngu@ctu.edu.vn
#
These authors contributed equally to this work

ABSTRACT
In the present study, the association between a 24-bp Insertion-Deletion (Indel) in the prolactin receptor gene (PRL), and
the polymorphic site NC_006091.5: c.25.144.C>A(T) in the melatonin receptor gene (MTNR-1C) was investigated in
Japanese laying quails. In total, 396 female and 132 male quails were selected for the experiment. The number of eggs
was recorded daily in individual quails for egg production and other reproductive traits. Every four weeks, one egg per
quail was randomly selected for external and internal assessment. PCR-RFLP and PCR-SSCP were applied for
genotyping the Indel, and the single nucleotide polymorphism in PRL and MTNR-1C genes, respectively. The data was
continuously collected until 48 weeks of laying. For the effects of PRL Indel, it was found that DD genotyped quails
provided a slightly higher egg yield in the first 24 laying weeks (155.1 eggs/quail); however, when the quails reached 36
and 48 weeks of laying, the difference in egg production was clear (P<0.05) in quails with II genotype (213.0 and 281.9
eggs/quail, respectively). In the MTNR-1C gene, six genotypes were detected at the polymorphic site, of which AA and
AT genotyped quails were better for egg yield in comparison with those baring other genotypes. For egg quality, only
albumen weight and albumen ratio differed from quails with II genotype to those of other quails in the 1-24 laying
weeks. In the later period (25-48 weeks), yolk index was the highest in quails baring DD genotype. For the MTNR-1C
polymorphism, the differences (P<0.05) were found in traits related to albumen and yolk; however, no genotype really
stands out in terms of egg quality. In conclusion, the two polymorphisms studied may be utilized in breeding programs to
enhance egg yield and egg quality traits in Japanese quails of which the PRL (II) and MTNR-1C (AA) alleles are
suggested.
Key words: Association, egg traits, Japanese quails, MTNR-1C, PRL
Published first online March 31, 2021 Published online Nov. 20, 2021.

INTRODUCTION increase of prolactin secretion leads to the regression of

ovarian functions and is of crucial importance to egg
Meat and eggs originating from poultry production and brooding behavior (Sharp et al. 1984,
production have traditionally been the main food supply Shimada et al. 1991). In Japanese quails, the molecular
for human consumption, thus it is a necessary large characterization of the PRL gene polymorphism is rare
industry all over the world. In addition to industrial and the majority of the information recorded is restricted
production for chickens, Japanese quail (Coturnix to indigenous or commercial chickens (Lotfi et al. 2013).
coturnix japonica) has gained much preference in recent In the 5'-flanking region of chicken PRL gene there was
years. In fact, large-scale production of Japanese quails an abundance of single-nucleotide polymorphisms (SNP)
for commercial purposes has been widely established in (Liang et al. 2006), and the 24-bp Indel was considerably
several regions (Narinc et al. 2013). In quails, the correlated with broodiness and egg yield (Cui et al. 2006,
selection of individuals based on genotypic information Jiang et al. 2005), which indicates its usefulness as a
related to reproductive traits is preferred (Deeb and molecular marker for egg production.
Lamont 2002). The research on candidate genes and their Melatonin (N-acetyl-5-methoxytryptamine)
impact on economic value traits have laid the foundation secreted from the pineal gland is an important hormone
for the use of molecular markers in breeding (Kulibaba that is involved in many functions in mammals, including
and Podstreshnyi 2012). feeding patterns and changing patterns of reproduction
Prolactin is a peptide hormone secreted by the (Adachi et al. 2002, He et al. 2016). Also, melatonin
anterior pituitary gland and it has a variety of roles for helps activate many receptor mechanisms in the ovary
biological functions in all vertebrates. In birds, the and various cells, including theca and granulosa cells

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(Ayre et al. 1992, Soares et al. 2003). Some studies have internal egg properties, album and yolk were evaluated
shown that the presence of melatonin in follicular fluid using a tripod micrometer and a digital caliper. The yolk
and its levels are positively correlated with the quality weight was recorded and the proportion was expressed in
and maturity of oocytes (Nakamura et al. 2003, Shi et al. accordance with egg weight. Albumen weight was
2009, Tian et al. 2014). Recently, one of the ovarian determined by the difference of the yolk and the shell
melatonin receptors, melanin receptor 1C (MTNR-1C) weight from the whole egg weight. The yolk index was
has been defined in chickens, and its expression has estimated from the ratio of yolk height to yolk width and
shown a direct contribution to the reproductive process of the albumen index was calculated as the ratio of thick
hens and is therefore marked as a candidate gene for albumen to its thick diameter. In addition, the Haugh unit
chicken reproduction (Li et al. 2013). A similar mutation (HU) was determined from the formula of HU = 100 x
was also detected in Noi chickens, a native chicken breed log (AH + 7.57 – 1.7 x EW0.37), in which AH stands for
in Vietnam (Vu and Ngu 2016). albumen height (mm) and EW represents egg weight (g)
The current study was conducted to identify the (Nasr et al. 2016, Sari et al. 2012).
polymorphisms of PRL and MTNR-1C genes and further
DNA extraction and genotyping: Genomic DNA
investigate their association with egg production and egg
isolated from quails’ feathers (Bello et al. 2001) was
quality traits in Japanese quails.
subjected to amplification in a thermal cycler. In the
prolactin gene (Genbank number AB011438.2), the
MATERIALS AND METHODS primer pair was designed for amplifying the fragment in
chickens (Cui et al. 2006). The primer sequences were
Experimental quails: The quails studied were selected 5’-TTT AAT ATT GGT GGG TGA AGA GAC A-3’
from a breeding farm in Tien Giang province. In total, (forward) and ATG CCA CTG ATC CTC GAA AAC TC
396 laying quails and 132 male quails were selected. The (reverse). The polymerase chain reaction (PCR) was
birds were fed a grower diet (ME: 2,950 kcal/kg; CP: performed with an annealing temperature of 54 oC and the
24%) from 0 to 6 weeks, and a laying diet (ME: 2,850 subsequence genotyping by PCR-RFLP (Restriction
kcal/kg; CP: 21%) for the whole experiment. Clean water Fragment Length Polymorphism) of all birds was
was provided at all times. During the laying period, the completed by following the protocol described by Cui et
female quails were kept in individual compartments for al. (2006).
egg collection and the male was alternatively introduced For the MTNR-1C gene, forward (5’-
every 4 hours per day with the ratio of 1 male to 3 TGCCAGATAAGTGGGTTCCT-3’) and reverse (5’-
females. All quails were vaccinated against common AGCGTCCACGTCAGACAGAT-3’) primers were
diseases before the experimental period of 48 laying designed by the primer3 tool (Untergrasser et al. 2012) to
weeks. In detail, Newcastle virus vaccine was amplify a fragment of 164 base pairs (Genbank number
administered on day 1st and day 21st via eye drop and NC_006091.5). Polymerase chain reactions were carried
repeated every 3 months orally, and the vaccine against out with the following conditions: 95°C for 5 min,
coccidiosis was applied on day 5th and day 30th through followed by 35 cycles at 94°C for 30 s, 55°C for 45 s,
drinking water. In addition, vaccines for chronic 72°C for 50 s, and ended with an extended cycle at 72°C
respiratory disease (CRD) and salmonella were given on for 5 min. Each PCR contained 10 µL, including 50 ng of
day 12th via drinking water and H5N1 vaccine was used genomic DNA, 0.25 M of each primer, 0.25 M of each
on day 15th by intramuscular injection. dNTP, 1x PCR buffer, and 1U/µL Taq DNA polymerase.
Egg production and egg quality measurements: Eggs The polymorphic site was identified by sequencing with
were collected daily at 4 P.M. and numbered to monitor Sanger’s method. For genotyping, the PCR-SSCP
individual yield. The eggs were then marked and (Single-Strand Conformation Polymorphism) technique
gathered in a storage room for 4 days before an was applied. In brief, the mixture of 8 µl of PCR product
incubation period of 17 days in an incubator. The and 10 µl loading buffer was heated to 95°C and
following parameters were recorded as phenotypic data maintained for 10 minutes to separate the DNA strands.
of reproductive capacity: total egg yield (per bird basis), The suspension was quickly transferred to ice and kept at
total egg hatched, number of fertilized eggs and hatched -20°C for 10 minutes. Before electrophoresis, the gel was
chicks, laying rate and hatching rate. cooled at 7°C for 30 minutes. The DNA mixture was
For evaluation of the egg quality traits, one loaded on gel and run at 10°C for 6 hours at 30 W. After
egg/quail was collected every four weeks with a total of that, the gel was stained with 250 ml of TAE 1X solution
12 eggs/quail used for quality assessment for the whole containing 25 µl of ethidium bromide (25 mg) for 30
experimental period. Eggs were weighed using an minutes and visualized under UV light of the gel imaging
electronic scale. After breaking the outer shells, eggshells system (Gel Logic 212 - Kodak).
were washed and dried for 24 hours before weighing to Statistical analysis: The association between genotype
obtain the ratio between eggshells and weight. For and egg yield and egg quality traits was analyzed based

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on the General Linear Model of Minitab software version (P=0.061) in quails with DD genotypes. In the next
16.2.1 (Minitab 2010), Yij = µ + Gi + ξij (where Yij: period, when the quails reached nine months of laying,
traits observed; μ: general mean, Gi: influence of egg yield was the highest in birds carrying the II
genotype; ξij: random error). Data is presented as Least genotype (213 eggs/bird; P<0.05). Due to similar
square mean ± Standard error. fertilized and hatching rates, the number of fertilized eggs
and hatched chicks was also higher in type II quails. The
RESULTS difference was even more apparent when eggs were
collected up to 48 weeks of laying with the highest egg
Total egg yield was associated with the insertion production in birds baring II genotype, while those with
or deletion of nucleotides in the prolactin gene depending DD genotype were the lowest (P<0.01). However, it is
on laying stages (Table 1). Considering only in 12 laying worth considering that quails with DD genotype
weeks, all reproductive parameters remained similar presented only at 3.9% in the population, whereas quails
(P>0.05); however, a tendency of difference occurred at carrying II genotype occupied 45.1% in the population
six months of egg yield with a slightly higher rate studied (396 individuals).

Table 1. Association between PRL polymorphism with egg production in different laying periods.

Genotypes
Parameters P
II (178) ID (n=203) DD (n=15)
1-12 laying weeks
Total egg yield 71.3±0.6 70.6±0.5 73.2±2.1 0.364
Laying rate (%) 84.8±0.7 84.0±0.6 87.2±2.5 0.364
Total egg hatched 67.3±0.6 66.9±0.5 70.0±2.1 0.333
Fertilized eggs 57.7±0.7 56.7±0.6 66.3±2.3 0.121
Fertilized rate (%) 85.7±0.6 84.7±0.6 87.2±2.2 0.312
Hatched chick number 53.2±0.7 52.1±0.6 56.9±2.4 0.125
Hatching rate (%) 92.0±0.4 91.7±0.4 92.8±1.5 0.731
1-24 laying weeks
Total egg yield 148.4±1.0 146.6±0.9 155.1±3.8 0.061
Laying rate (%) 88.3±0.6 87.2±0.6 92.3±2.3 0.061
Total egg hatched 140.6±1.0 139.4±0.9 147.8±3.8 0.087
Fertilized eggs 122.5ab±1.3 119.0b±1.2 133.7a±5.1 0.007
Fertilized rate (%) 87.1a±0.7 85.3b±0.6 90.6a±2.6 0.039
Hatched chick number 111.0ab±1.7 106.7b±1.6 123.0a±6.4 0.016
Hatching rate (%) 89.9±0.6 88.9±0.6 91.8±2.3 0.253
1-36 laying weeks
Total egg yield 213.0a±2.4 204.7b±2.2 205.2ab±9.0 0.034
Laying rate (%) 84.5a±0.9 81.2b±0.9 81.4ab±3.6 0.034
Total egg hatched 199.9a±2.3 192.4b±2.2 193.3a±8.8 0.047
Fertilized eggs 172.4a±1.9 165.0b±1.8 171.2ab±7.3 0.019
Fertilized rate (%) 86.6±0.5 86.2±0.5 88.6±2.0 0.447
Hatched chick number 150.3±1.8 145.1±1.6 151.5±6.7 0.056
Hatching rate (%) 87.4±0.5 88.1±0.5 90.2±2.0 0.298
1-48 laying weeks
Total egg yield 281.9a±2.8 267.3b±3.9 244.8b±15.0 0.007
Laying rate (%) 83.9a±1.4 79.6b±1.2 72.9b±4.5 0.007
Total egg hatched 272.9a±4.2 258.3b±3.9 235.8b±15.0 0.007
Fertilized eggs 234.9a±3.3 221.0b±3.2 207.6ab±11.9 0.003
Fertilized rate (%) 86.7±0.5 86.1±0.5 88.5±1.8 0.373
Hatched chick number 203.5a±2.8 193.1b±2.6 187.0ab±9.9 0.014
Hatching rate (%) 86.9±0.5 87.8±0.5 90.1±1.9 0.174
a,b,c Values within a row with different superscripts are significantly different (P<0.05)

At the polymorphic site of the MTRN-1C gene, when the yield was counted in 12 weeks or until 24
the effects of the genotype were found in egg production weeks of laying (P<0.01), but in the later stage, the

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difference was non-significant (Table 2). Superior egg similar, with the exception of yolk index, in which the
yield was found in quails of AA genotype while those of highest was presented in DD genotype quails (P<0.05).
in TT genotyped quails were inferior (153.4 vs. 144.6 In the first 6 months of laying, many egg quality
eggs/24 laying weeks, respectively). A similar trend was characteristics were associated with the MTRN-1C
also found in several fertilized eggs and hatched chicks. polymorphism, especially egg weight and other traits
In Table 3, egg quality is presented for different related to yolk and albumen (Table 4). The genotype CC
laying periods (1-24 weeks and 25-48 weeks). Among the showed lower egg weight than both CT and TT
external and internal egg traits, albumen weight and genotypes, which was also observed for the yolk index.
albumen ratio were found to differ (P<0.01) in the first 6 Another internal trait, HU was the highest in birds with
months of laying, with the highest values being in quails TT genotype. However, in the later stage, only yolk ratio
of II genotype (6.6 g and 54.3%, respectively). In the was affected by genotype (P<0.05).
later stage (25-48 weeks), most of the parameters were

Table 2. Association between MTNR-1C polymorphism with egg production for extended laying periods.

Genotypes
Parameters P
AA (n=40) AC (n=64) AT (n=101) CC (n=25) CT (n=80) TT (n=64)
1-12 laying weeks
Total egg yield 72.4ab±0.6 72.7a±0.7 67.6c±1.4 71.4abc±1.0 71.6abc±0.7 70.1bc±0.6 0.002
Laying rate (%) 86.2 ±0.8
ab
86.5 ±0.8
a
80.4 ±1.7
c
85.0abc±1.1 85.2abc±0.9 83.4bc±0.7 0.002
Total eggs hatched 68.6 ±0.6
ab
68.8 ±0.7
a
63.8 ±1.4
c
67.6abc±0.9 67.7abc±0.7 66.3bc±0.6 0.002
Fertilized eggs 58.8±0.8 57.6±0.9 55.1±1.8 57.7±1.2 57.3±0.9 57.1±0.7 0.439
Fertilized rate (%) 85.8±0.8 83.6±0.9 86.3±1.8 85.3±1.2 84.6±1.0 86.0±0.7 0.342
Hatched chick number 54.6±0.9 52.7±1.0 51.0±2.0 53.1±1.4 52.5±1.0 52.7±0.8 0.477
Hatching rate (%) 92.6±0.7 91.0±0.7 92.4±1.5 91.9±1.0 91.5±0.7 92.0±0.6 0.661
1-24 laying weeks
Total egg yield 153.4a±1.3 148.9ab±1.5 153.9a±2.8 149.7ab±2.1 149.3ab±1.5 144.6b±1.2 0.000
Laying rate (%) 91.3a±0.8 88.7ab±0.9 91.6a±1.7 89.1ab±1.3 88.9ab±0.9 86.1b±0.7 0.000
Total eggs hatched 144.9 ±1.3 140.6 ±1.5 145.4 ±2.7 141.4 ±2.1 140.9ab±1.5
a ab a ab
136.3b±1.2 0.000
Fertilized eggs 126.1a±1.9 118.4ab±2.1 127.1ab±3.9 119.6ab±2.9 121.8ab±2.1 118.6b±1.7 0.019
Fertilized rate (%) 87.0±1.0 84.0±1.1 87.4±2.0 84.5±1.5 86.2±1.1 86.9±0.8 0.211
Hatched chick number 114.8±2.4 105.7±2.7 117.6±5.1 106.0±3.8 109.6±2.7 107.4±2.1 0.052
Hatching rate (%) 90.5±0.9 87.9±1.0 92.1±1.9 87.7±1.4 89.3±1.0 90.0±0.8 0.196
1-36 laying weeks
Total egg yield 210.6±3.6 205.0±3.8 208.4±7.0 214.9±5.5 206.5±4.0 206.6±3.2 0.686
Laying rate (%) 83.6±1.4 81.3±1.5 82.7±2.8 85.3±2.2 82.0±1.6 82.0±1.3 0.686
Total eggs hatched 199.0±3.6 193.5±3.9 196.9±7.1 203.5±5.5 194.9±4.0 195.4±3.2 0.705
Fertilized eggs 172.8±3.0 164.8±3.2 173.3±5.9 172.7±4.6 167.5±3.4 168.1±2.7 0.445
Fertilized rate (%) 87.3±0.8 85.7±0.8 88.2±1.5 85.2±1.2 86.4±0.9 86.1±0.7 0.456
Hatched chick number 153.5±2.7 146.2±2.9 152.7±5.3 150.8±4.1 147.7±3.0 147.3±2.4 0.405
Hatching rate (%) 89.1±0.7 89.0±0.8 88.5±1.5 87.3±1.1 88.3±0.8 87.8±0.7 0.065
1-48 laying weeks
Total egg yield 266.0±6.4 264.9±6.8 246.1±14.8 268.3±9.4 265.2±7.1 270.3±5.7 0.783
Laying rate (%) 79.2±1.9 78.9±2.0 73.3±4.4 79.9±2.8 78.9±2.1 80.5±1.7 0.783
Total eggs hatched 250.6±6.3 249.6±6.8 230.7±14.7 253.0±9.4 249.6±7.1 254.6±5.7 0.784
Fertilized eggs 261.8±5.1 213.2±5.4 204.3±11.8 214.9±7.5 213.4±5.6 218.3±4.5 0.892
Fertilized rate (%) 87.3±0.7 86.1±0.8 85.3±1.7 85.6±1.1 86.3±0.8 86.0±0.7 0.555
Hatched chick number 191.6±4.1 188.0±4.4 183.6±9.5 187.3±6.0 186.3±4.6 190.2±3.7 0.936
Hatching rate (%) 89.0±0.8 88.7±0.8 90.0±1.7 87.5±1.1 87.8±0.8 87.5±0.7 0.494
a,b,c
Values within a row with different superscripts are significantly different (P<0.05)

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Table 3. Association between PRL polymorphism and egg quality traits.

Genotype
Parameters P
II ID DD
1-24 laying weeks
Total eggs checked 1.068 1.218 108
Shape index 76.3±0.3 75.9±0.2 76.3±1.3 0.682
Egg weight (g) 12.2±0.1 12.0±0.1 12.2±0.3 0.240
Egg shell weight (g) 1.6±0.01 1.6±0.01 1.5±0.01 0.291
Albumen weight (g) 6.6a±0.1 6.3b±0.1 5.1c±0.4 0.000
Albumen height (mm) 4.1±0.03 4.1±0.02 4.1±0.1 0.554
Albumen width (mm) 41.2±0.8 44.6±0.7 45.9±3.3 0.842
Yolk weight (g) 3.9±0.02 3.8±0.02 3.8±0.1 0.156
Yolk height (mm) 11.0±0.1 10.9±0.03 11.1±0.2 0.081
Yolk width (mm) 23.4±0.2 23.4±0.1 23.6±0.7 0.953
Egg shell ratio (%) 12.9±0.1 12.9±0.1 12.3±0.4 0.347
Albumen ratio (%) 54.3a±0.7 52.1b±0.6 41.9c±3.0 0.000
Albumen index 9.5±0.1 9.5±0.1 9.1±0.5 0.780
Yolk ratio (%) 31.6±0.1 31.4±0.1 31.5±0.6 0.595
Yolk index 48.0±0.6 47.3±0.2 47.2±2.6 0.687
Haugh unit 87.0±0.2 86.9±0.1 86.9±0.7 0.849
25-48 laying weeks
Total eggs checked 1068 1218 108
Shape index 76.3±0.3 75.9±0.3 75.2±1.2 0.548
Egg weight (g) 11.8±0.1 11.8±0.1 11.7±0.3 0.831
Egg shell weight (g) 1.6±0.01 1.6±0.01 1.4±0.1 0.097
Albumen weight (g) 6.6±0.1 6.6±0.1 6.5±0.2 0.929
Albumen height (mm) 4.4±0.03 4.4±0.03 4.3±0.1 0.268
Albumen width (mm) 42.9±0.4 42.3±0.4 42.2±1.8 0.585
Yolk weight (g) 3.7±0.03 3.7±0.03 3.8±0.1 0.364
Yolk height (mm) 10.8±0.1 10.9±0.1 11.2±0.5 0.494
Yolk width (mm) 26.5±1.3 23.2±1.1 22.9±5.4 0.148
Egg shell ratio (%) 13.2±0.1 13.3±0.1 13.3±0.5 0.115
Albumen ratio (%) 55.3±0.2 55.6±0.2 54.9±1.0 0.677
Albumen index 10.4±0.2 10.7±0.1 10.4±0.6 0.287
Yolk ratio (%) 31.4±0.2 31.1±0.2 32.8±0.8 0.084
Yolk index 45.8b±0.4 46.7b±0.3 49.1a±1.2 0.033
Haugh unit 88.6±0.2 89.0±0.2 88.2±0.8 0.264
a,b,c Values within a row with different superscripts are significantly different (P<0.05)

Table 4. Association between MTNR-1C polymorphism with egg quality traits.

Genotypes
Parameters P
AA AC AT CC CT TT
1-24 laying weeks
Total eggs checked 486 120 384 216 612 426
Shape index (%) 76.7±0.5 75.9±0.6 75.7±1.1 76.4±0.8 75.9±0.6 75.6±0.5 0.727
Egg weight (g) 12.2ab±0.1 12.0ab±0.1 12.2ab±0.2 11.7b±0.1 12.3a±0.1 12.2a±0.1 0.025
Egg shell weight (g) 1.6±0.02 1.5±0.02 1.6±0.03 1.5±0.02 1.6±0.02 1.6±0.02 0.485
Albumen weight (g) 6.3±0.1 6.1±0.1 6.8±0.3 6.1±0.2 6.5±0.2 6.5±0.1 0.066
Albumen height (mm) 4.0b±0.04 4.2ab±0.04 4.1ab±0.01 4.1ab±0.1 4.1ab±0.04 4.2a±0.03 0.005
Albumen width (mm) 44.6±1.1 42.9±1.2 45.3±2.2 42.6±1.2 45.2±1.3 45.5±0.9 0.388
Yolk weight (g) 3.9±0.04 3.7±0.04 3.8±0.1 3.8±0.1 3.8±0.1 3.8±0.03 0.085
Yolk height (mm) 11.0abc±0.1 11.1a±0.1 11.1abc±0.1 10.7c±0.1 10.8bc±0.1 11.0ab±0.1 0.002
Yolk width (mm) 23.8a±0.3 23.6ab±0.3 23.6ab±0.5 24.0a±0.3 22.5b±0.3 23.2ab±0.2 0.003
Egg shell ratio (%) 12.8±0.1 12.9±0.1 12.8±0.3 13.1±0.2 12.7±0.2 13.0±0.1 0.525

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Albumen ratio (%) 52.1±1.1 51.6±1.1 56.3±2.1 51.9±1.4 53.3±1.2 52.8±0.9 0.429
Albumen index (%) 9.1ab±0.2 9.8a±0.2 9.1ab±0.3 9.6ab±0.2 9.1b±0.2 9.6ab±0.1 0.013
Yolk ratio (%) 31.9ab±0.2 31.1c±0.2 30.8bc±0.4 32.3a±0.3 30.7c±0.2 31.4bc±0.2 0.000
Yolk index 46.3ab±0.9 48.6ab±0.9 47.2 ±1.7
ab
44.8b±1.1 49.7a±1.0 47.9ab±0.7 0.015
Haugh unit 86.3b±0.2 87.3a±0.2 86.9ab±0.4 86.9ab±0.3 86.5ab±0.3 87.3a±0.2 0.004
25-48 laying weeks
Total eggs checked 486 120 384 216 612 426
Shape index (%) 11.8±0.1 11.8±0.1 11.8±0.2 11.5±0.1 11.9±0.1 11.8±0.1 0.254
Egg weight (g) 11.8±0.1 11.9±0.1 11.9±0.2 11.5±0.1 12.0±0.1 11.9±0.1 0.198
Egg shell weight (g) 1.5±0.02 1.6±0.02 1.6±0.04 1.5±0.03 1.6±0.02 1.6±0.02 0.351
Albumen weight (g) 6.5ab±0.1 6.6ab±0.1 6.6ab±0.1 6.3b±0.1 6.7a±0.1 6.6ab±0.1 0.044
Albumen height (mm) 4.4±0.1 4.4±0.1 4.5±0.1 4.4±0.1 4.3±0.1 4.4±0.04 0.145
Albumen width (mm) 42.4±0.6 42.6±0.6 44.0±1.2 41.8±0.8 43.3±0.7 42.5±0.5 0.654
Yolk weight (g) 3.7±0.04 3.7±0.04 3.7±0.09 3.7±0.06 3.7±0.07 3.7±0.04 0.697
Yolk height (mm) 10.7±0.2 11.2±0.2 10.8±0.3 10.7±0.2 10.8±0.2 10.7±0.1 0.249
Yolk width (mm) 28.9±1.8 23.5±1.9 23.7±3.6 23.2±2.4 23.6±2.1 23.4±1.5 0.205
Egg shell ratio (%) 13.1±0.2 13.4±0.4 13.0±0.3 13.3±0.2 13.2±0.2 13.2±0.1 0.797
Albumen ratio (%) 55.3±0.3 55.8±0.3 55.9±0.6 54.8±0.4 56.3±0.4 55.4±0.3 0.161
Albumen index (%) 10.6±0.2 10.6±0.2 10.4±0.4 10.7±0.3 10.2±0.3 10.5±0.2 0.687
Yolk ratio (%) 31.6ab±0.3 30.8ab±0.3 31.1ab±0.5 31.9a±0.3 30.5b±0.3 31.4ab±0.2 0.015
Yolk index 45.1±0.7 47.5±0.8 45.9±1.4 46.2±1.0 46.6±0.8 46.1±0.6 0.345
Haugh unit 89.1±0.3 89.0±0.3 89.6±0.6 89.1±0.4 88.0±0.3 88.7±0.2 0.081
a,b,c Values within a row with different superscripts are significantly different (P<0.05)

DISCUSSION Bagheri Sarvestani et al. (2013) that revealed genotype II

and ID had higher hatching rates, whereas genotype DD
In the present report, a 24-bp Indel and a point gave a lower rate.
mutation NC_006091.5: c.25.144.C>A(T) was detected There has been evidence showing the linkage
in PRL and MTNR-1C genes of Japanese quails, between the PRL polymorphic site with egg production.
respectively. The polymorphisms showed certain effects Jiang et al. (2005) informed that chickens with
on egg production and internal egg quality traits, which homozygote insertion (II genotype) could reduce
may indicate that they can be of interest in the selection prolactin expression leading to no broodiness in chickens.
of reproductive traits in breeding programs. Also, the report from Shiue et al. (2006) revealed that
In the PRL gene, the presence of the 24-bp Indel mRNA expression of the prolactin gene was significantly
with three genotypes, including II (154 bp), ID (154 and higher in low egg-producing chickens, which indicates
130 bp) and DD (130 bp), was found. The results also that egg production was related to prolactin mRNA
support the statement that in non-commercial breeds of expression. Then, a rise of prolactin level in plasma may
chickens, the “I” allele frequency varies from low to induce incubation behavior and then terminate laying
medium (Begli et al. 2010, Cui et al. 2006, Rashidi et al. (Sockman et al. 2000) and thus decrease egg production
2012), while in the commercial layer, this allele is (Reddy et al. 2002). The elevated levels of prolactin
commonly found. Besides, these results correlate decrease the egg clutch lengths by increasing the inter-
favorably with the reports on chickens (Bagheri sequence pauses between the sequences of egg lay,
Sarvestani et al. 2013), fowls (Begli et al. 2010), and give particularly in native birds (Reddy et al. 2002, Reddy et
further support to the conclusion found for quails (Lotfi al. 2006). Prolactin also plays a significant role in
et al. 2013, Yousefi et al. 2012). It has been demonstrated decreasing the number of follicles in the ovary, thus
that II genotype has a positive effect on egg production affecting egg production. In that respect, broodiness was
efficiency compared with effects from ID and DD adversely correlated with egg production and finally
genotypes (Lotfi et al. 2013). In the present study, stimulated the reduction of egg production in domestic
Japanese quails possessing II genotype had higher egg fowl (March et al. 1994).
production, which is also in agreement with the report of It was also noted that, in the first months of egg-
Begli et al. (2010) in Fars native chickens. Supportably, laying, the productivity of DD genotyped quails tended to
the increase of allele I in the population has been reported be higher, then decreased rapidly during the 7-9 month
to eliminate the incubation behavior in local Papua hens period. In reality, if quails up to 24 weeks of laying are
(Lumatauw and Mu’in 2016). However, the current used for egg production, then it is reasonable to
results were in contrast with the previous report by incorporate the selection of this genotype into breeding,
yet with longer laying periods, productivity will decrease

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rapidly. However, in this study, there were only 15 there are differences between quails that carry different
individuals with genotypes DD out of a total of 396 genotypes, no genotype really stands out in egg quality.
individuals, so it is not possible to have a precise Quails carrying AA genotype gave an average egg mass
conclusion about the benefits and disadvantages of this compared with those with other genotypes, and there is
genotype. Meanwhile, II genotype resulted in relatively no difference in other indicators related to albumen and
stable egg production and a slow reduction in laying rate. yolk. However, this is a homogeneous genotype for high
Therefore, when considering the effectiveness of quails yield, so it should be noted for breeding purposes.
up to 48 weeks of laying, this would be the best choice in
Conclusion: Altogether, it can be concluded that the two
terms of productivity and flock development.
polymorphisms studied may be used in breeding
In the present study, a novel polymorphic site
programs to improve egg production, and to some extent,
was detected in MTNR-1C gene and it was found to
egg quality traits in Japanese quails, of which the II
associate with egg yield and some egg quality traits.
genotype in PRL gene and the homozygous AA genotype
According to Sundaresan et al. (2009), the MTNR-1C
in MTNR-1C genes are appropriate for selection.
gene is highly expressed in the follicular granulocytes;
thus, it is thought to play an important role in egg Acknowledgments: This research was funded by the
formation, thereby affecting poultry egg production. The Vietnam National Foundation for Science and
study by Li et al. (2015) showed that the expression of Technology Development (NAFOSTED) under grant
the MTNR-1C gene was related to chicken egg number 106-NN.05-2013.12.
production, and the authors demonstrated that chickens
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