This study was to detect the expression of heart fatty acid-binding protein (H-FABP) and adipocyte fatty acid-binding protein (A-FABP) gene mRNA in different tissues of Rugao and Luyuan chickens at 56 d and 120 d by real-time fluorescence quantitative reverse transcription polymerase-chain reaction (FQ-RT-PCR). The primers were designed according to the sequences of HFABP, A-FABP and GAPDH genes in Gallus gallus, which were used as target genes and internal reference gene, respectively. The levels of H-FABP and A-FABP gene expression were detected by SYBR Green I FQ-RT-PCR. The relative H-FABP and A-FABP gene mRNA expression level was calculated with 2-$^{{\Delta}Ct}$. Melting curve analysis showed a single peak of three genes. Intramuscular fat (IMF) content in breast muscle and leg muscle of the two chicken breeds at 120 d was higher than at 56 d. IMF content in breast muscle and leg muscle at 56 d and 120 d in Luyuan was significantly higher than in Rugao, however, abdominal fat of Luyuan was significantly lower than that of Rugao. The relative H-FABP gene mRNA expression level in cardiac muscle was the highest in both chicken breeds. The relative H-FABP and A-FABP gene expression of different tissues in Luyuan was higher than in Rugao. H-FABP gene mRNA expression had a negative effect on IMF of leg and breast muscles, and was significantly negatively correlated with IMF content. The relative A-FABP gene mRNA level in abdominal fat was higher than in liver. The A-FABP gene mRNA was not expressed in leg, breast and cardiac muscles. A-FABP gene mRNA expression level was significantly positively correlated with abdominal fat and had a significant effect on abdominal fat but not IMF content.
Raillietina species are prevalent in domestic chickens (Gallus gallus domesticus) in Phayao province, northern Thailand. Their infection may cause disease and death, which affects the public health and economic situation in chicken farms. The identification of Raillietina has been based on morphology and molecular analysis. In this study, morphological observations using light (LM) and scanning electron microscopies (SEM) coupled with molecular analysis of the internal transcribed spacer 2 (ITS2) region and the nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1) gene were employed for precise identification and phylogenetic relationship studies of Raillietina spp. Four Raillietina species, including R. echinobothrida, R. tetragona, R. cesticillus, and Raillietina sp., were recovered in domestic chickens from 4 districts in Phayao province, Thailand. LM and SEM observations revealed differences in the morphology of the scolex, position of the genital pore, number of eggs per egg capsule, and rostellar opening surface structures in all 4 species. Phylogenetic relationships were found among the phylogenetic trees obtained by the maximum likelihood and distance-based neighbor-joining methods. ITS2 and ND1 sequence data recorded from Raillietina sp. appeared to be monophyletic. The query sequences of R. echinobothrida, R. tetragona, R. cesticillus, and Raillietina sp. were separated according to the different morphological characters. This study confirmed that morphological studies combined with molecular analyses can differentiate related species within the genus Raillietina in Thailand.
Meydan, Hasan;Jang, Cafer Pish;Yildiz, Mehmet Ali;Weigend, Steffen
Asian-Australasian Journal of Animal Sciences
/
v.29
no.11
/
pp.1547-1554
/
2016
To assess genetic diversity and maternal origin of Turkish and Iranian native chicken breeds, we analyzed the mtDNA D-loop sequences of 222 chickens from 2 Turkish (Denizli and Gerze) and 7 Iranian (White Marandi, Black Marandi, Naked Neck, Common Breed, Lari, West Azarbaijan, and New Hampshire) native chicken breeds, together with the available reference sequences of G. gallus gallus in GenBank. The haplotype diversity was estimated as $0.24{\pm}0.01$ and $0.36{\pm}0.02$ for Turkish and Iranian populations, respectively. In total, 19 haplotypes were observed from 24 polymorphic sites in Turkish and Iranian native chicken populations. Two different clades or haplogroups (A and E) were found in Turkish and Iranian chickens. Clade A haplotypes were found only in White Marandi, Common Breed and New Hampshire populations. Clade E haplotypes, which are quite common, were observed in Turkish and Iranian populations with 18 different haplotypes, of which Turkish and Iranian chickens, Clade E, haplotype 1 (TRIRE1) was a major haplotype with the frequency of 81.5% (181/222) across all breeds. Compared to red jungle fowl, Turkish and Iranian chicken breeds are closely related to each other. These results suggest that Turkish and Iranian chickens originated from the same region, the Indian subcontinent. Our results will provide reliable basic information for mtDNA haplotypes of Turkish and Iranian chickens and for studying the origin of domestic chickens.
The variability of mtDNA hypervariable segment I (HVS I) sequences was investigated in a total of 48 birds belonging to 12 Chinese native chicken breeds. Sixteen haplotypes were identified from 35 polymorphic nucleotide sites which accounted for 6.4% of a sequenced 544 bp fragment. Diversity analysis of the haplotypes showed that Tibetan, Langshan and Henan cockfight chicken had only one haplotype, while ancient haplotypes existed in Taihe silky and Chahua chicken. Phylogenetic analysis of the haplotypes suggested that Chinese native chicken breeds shared 5 maternal lineages and some breeds would share the same maternal lineage, regardless of their external features and ecological types. Both divergent and phylogenetic analysis of the haplotypes indicated the close genetic relationships between the Chinese native chicken breeds and G. g. gallus and G. g. spadiceus from different areas, which implied that G. g. gallus and G. g. spadiceus were the original ancestors of the Chinese native chicken breeds.
Molee, A.;Kongroi, K.;Kuadsantia, P.;Poompramun, C.;Likitdecharote, B.
Asian-Australasian Journal of Animal Sciences
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v.29
no.1
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pp.29-35
/
2016
The aim of the present study was to investigate the effect of single nucleotide polymorphisms in the major histocompatibility complex (MHC) class II gene on resistance to Newcastle disease virus and body weight of the Thai indigenous chicken, Leung Hang Khao (Gallus gallus domesticus). Blood samples were collected for single nucleotide polymorphism analysis from 485 chickens. Polymerase chain reaction sequencing was used to classify single nucleotide polymorphisms of class II MHC. Body weights were measured at the ages of 3, 4, 5, and 7 months. Titres of Newcastle disease virus at 2 weeks to 7 months were determined and the correlation between body weight and titre was analysed. The association between single nucleotide polymorphisms and body weight and titre were analysed by a generalized linear model. Seven single nucleotide polymorphisms were identified: C125T, A126T, C209G, C242T, A243T, C244T, and A254T. Significant correlations between log titre and body weight were found at 2 and 4 weeks. Associations between single nucleotide polymorphisms and titre were found for C209G and A254T, and between all single nucleotide polymorphisms (except A243T) and body weight. The results showed that class II MHC is associated with both titre of Newcastle disease virus and body weight in Leung Hang Khao chickens. This is of concern because improved growth traits are the main goal of breeding selection. Moreover, the results suggested that MHC has a pleiotropic effect on the titre and growth performance. This mechanism should be investigated in a future study.
Many consumers are increasingly concerned about the meat they eat, and accurate labelling is important due to public health, economic and legal concerns. Meat species adulteration is a common problem in the retail markets. In this study, a TaqMan$^{(R)}$ quantitative real-time polymerase chain reaction (PCR) assay was applied for its ability to quantify chicken meat, which was not indicated on the label, in 79 commercial pork products (ham, sausages, bacon and ground meat) producted by 10 different manufacturers. The amplification efficiency was 82.05% and the square regression coefficient ($R^2$) was 0.995. PCR results showed that 38.6% of ham samples, 50.0% of sausages samples, and 50.0% of ground meat samples were contaminated with chicken residuals, while the bacon samples were not contaminated with chicken residuals. Only twelve pork products of one of the manufacturers were in accordance with indicated in their labels. The PCR assay reported in this work could be particularly useful in inspection programs to verify the food labelling of commercial processed meats and to gain consumers' trust.
Objective: Internal organs indirectly affect economic performance and well-being of animals. Study of internal organs during later layer period will allow full utilization of layer hens. Hence, we conducted a genome-wide association study (GWAS) to identify potential quantitative trait loci or genes that potentially contribute to internal organ weight. Methods: A total of 1,512 chickens originating from White Leghorn and Dongxiang Blue-Shelled chickens were genotyped using high-density Affymetrix 600 K single nucleotide polymorphism (SNP) array. We conducted a GWAS, linkage disequilibrium analysis, and heritability estimated based on SNP information by using GEMMA, Haploview and GCTA software. Results: Our results displayed that internal organ weights show moderate to high (0.283 to 0.640) heritability. Variance partitioned across chromosomes and chromosome lengths had a linear relationship for liver weight and gizzard weight ($R^2=0.493$, 0.753). A total of 23 highly significant SNPs that associated with all internal organ weights were mainly located on Gallus gallus autosome (GGA) 1 and GGA4. Six SNPs on GGA2 affected heart weight. After the final analysis, five top SNPs were in or near genes 5-Hydroxytryptamine receptor 2A, general transcription factor IIF polypeptide 2, WD repeat and FYVE domain containing 2, non-SMC condensin I complex subunit G, and sonic hedgehog, which were considered as candidate genes having a pervasive role in internal organ weights. Conclusion: Our findings provide an understanding of the underlying genetic architecture of internal organs and are beneficial in the selection of chickens.
Objective: In Japan, approximately 50 breeds of indigenous domestic chicken, called Japanese native chickens (JNCs), have been developed. JNCs gradually became established based on three major original groups, "Jidori", "Shoukoku", and "Shamo". Tosa-Jidori is a breed of Jidori, and archival records as well as its morphologically primitive characters suggest an ancient origin. Although Jidori is thought to have been introduced from East Asia, a previous study based on mitochondrial D-loop sequences demonstrated that Tosa-Jidori belongs to haplogroup D, which is abundant in Southeast Asia but rare in other regions, and a Southeast Asian origin for Tosa-Jidori was therefore suggested. The relatively small size of the D-loop region offers limited resolution in comparison with mitogenome phylogeny. This study was conducted to determine the phylogenetic position of the Tosa-Jidori breed based on complete mitochondrial D-loop and mitogenome sequences, and to clarify its evolutionary relationships, possible maternal origin and routes of introduction into Japan. Methods: Maximum likelihood and parsimony trees were based on 133 chickens and consisted of 86 mitogenome sequences as well as 47 D-loop sequences. Results: This is the first report of the complete mitogenome not only for the Tosa-Jidori breed, but also for a member of one of the three major original groups of JNCs. Our phylogenetic analysis based on D-loop and mitogenome sequences suggests that Tosa-Jidori individuals characterized in this study belong to the haplogroup D as well as the sub-haplogroup E1. Conclusion: The sub-haplogroup E1 is relatively common in East Asia, and so although the Southeast Asian origin hypothesis cannot be rejected, East Asia is another possible origin of Tosa-Jidori. This study highlights the complicated origin and breeding history of Tosa-Jidori and other JNC breeds.
Owing to its excellent nutritional value, eggs are among the most important components of the human diet. Gender and environmental factors, such as feed composition, may alter the nutritional profile and quality of eggs. Feed additives have recently been used to enhance the health and productivity of hens, which has resulted in the production of higher-quality eggs. The fungus Cordyceps militaris, a well-established source of traditional medicines, contains potential bioactive metabolites, which prompted us to examine the effects of C. militaris-supplemented diets on the quality of hens' eggs. The hens of two species (Gallus gallus domesticus and Araucana) were fed with one of three different diets: a control diet and diets supplemented with 2% or 5% of C. militaris. Egg quality was determined by measuring the Haugh Unit, yolk color, and shell thickness. In addition, egg and shell densities together with the ratio of yolk to albumen were calculated. Eggshell thickness and yolk color were both enhanced by the addition of C. militaris, whereas Haugh Unit values were somewhat reduced. Egg size, eggshell weight, and yolk and albumen production were all enhanced by C. militaris supplementation. Notably, in hens fed the 2% C. militaris-supplemented diet, enhancement was more evident in the yolk than in the albumen. The overall quality of the egg yolk was enhanced when 2% C. militaris was added to the hens' diet, which led to increases in both yolk color and quantity. Eggshell thickness and weight were also higher among eggs laid by hens fed the supplemented diets. Although these effects differed depending on the chicken species, we established that, in general, C. militaris contributes to improving egg quality.
Objective: This study aimed to investigate the effects of corticotropin-releasing factor (CRF) on the feed intake of broiler chickens and explore its influencing mechanism. Methods: The study included two trials. In trial 1, 32 male broiler chickens (Arbor Acres, Gallus gallus domesticus) were given ventricle buried tubes, and they were allowed to recover for 3 days. At 8:00 AM, intracerebroventricular (ICV) injection with CRF or normal saline was performed in 10-day-old broiler chickens, which were divided into the 5, 10, and 20 ㎍ and control (normal saline) groups according to the dose of CRF injection. In trial 2, chickens were divided into the 10 ㎍ and control group (physiological saline) to repeat trial 1. Results: Results of trial 1 showed that the cumulative amount of feed intake in the 10 or 20 ㎍ groups was considerably lower than that of the control group after ICV injection with CRF. The lowest amount of feed intake was obtained with the addition of 10 ㎍ of CRF. In trial 2, the expression of ghrelin in the hypothalamus injected with 10 ㎍ of CRF increased significantly, but the expression of ghrelin in various sections of the small intestine considerably decreased. The expression of CRF receptor subtypes 1 (CRFR1) in the hypothalamus and some parts of the small intestine remarkably increased, and the expression of CRF receptor subtypes 2 (CRFR2) increased only in the duodenum, whereas the expression of growth hormone secretagogue receptor (GHSR-1α) in the jejunum and ileum increased considerably after ICV injection of 10 ㎍ of CRF. Conclusion: The CRF at 10 ㎍ increased ghrelin expression in the hypothalamus and CRFR1 expression in the small intestine, and this phenomenon was related to the suppressed feed intake of broiler chickens.
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