Survivability,immunity, growth and production traits in indigenous and White Leghorn breeds of chicken

ABSTRACT

1. The survivability, innate and adaptive immunity, growth and production traits up to 72 weeks of age were determined in Ghagus, Nicobari (unimproved indigenous) and White Leghorn (WLH) breeds and the study investigated links between innate and adaptive immunity and survivability and production traits.

2. At 20 and 40 weeks of age, there was a significant effect of breed on innate immunity assessed by measuring titres of natural antibody (NAb) binding to rabbit red blood cells (RRBC) and adaptive immunity assessed by measuring specific antibody titre (SpAb) to Newcastle disease virus.

3. Highest survivability was in WLH (91.6%) followed by Nicobari (87.1%) and Ghagus (82.9%) breeds. Growth traits at different ages were higher (P< 0.001) in Ghagus followed by WLH and Nicobari breeds. Egg production up to 72 weeks was higher (P < 0.001) in WLH followed by Nicobari and Ghagus breeds, whereas egg weight at different ages was higher (P < 0.001) in WLH than Ghagus and Nicobari breeds.

4. NAb titres measured at 20 weeks were significantly (P = 0.002) associated with the survivability of hens during 20 to 72 weeks of age. Breed-wise analysis showed a significant (P = 0.019) association between NAb titres at 20 weeks and survivability in the Ghagus breed. Furthermore, NAb titres at 20 weeks were higher in hens which survived to 72 weeks compared with those that died (P = 0.002).

5. Measuring NAb titres to RRBC is quick, economical and simple. This method has potential to be used in a breeding programme to increase survivability of laying hens.     

甜菜碱对育肥猪生长性能和PRKAG3基因相对表达量的影响

为了研究甜菜碱对育肥猪生长性能和PRKAG3基因表达量的影响,试验采用单因子试验设计,选用80 kg左右的健康杜长大三元杂交猪60头,随机分成4组,每组3个重复,每个重复5头猪,在日粮中添加甜菜碱0,500,1 000,1 500 mg/kg,试验期为41 d,试验结束后随机选择(每组随机选择3头)12头猪进行屠宰测定。结果表明:与对照组相比,日粮中添加甜菜碱对育肥猪的日增重、日采食量、料重比均有明显影响,可以不同程度地提高日增重和采食量,降低料重比,其中以添加1 500 mg/kg甜菜碱(4组)的效果最为明显;日粮中添加甜菜碱可以不同程度地提高育肥猪PRKAG3基因的表达量,其中以1 500 mg/kg甜菜碱(4组)的效果最为明显。说明甜菜碱可以提供甲基供体,节约蛋氨酸和胆碱,从而提高育肥猪的生长性能。     

PRKAG3基因在骨骼肌中的营养代谢调节作用及其对肉质的影响

PRKAG3是近年来确定的一个影响猪肉pH、肉色以及系水力的主效基因,其突变R200Q与过量肌糖原含量有关,被认为是汉普夏猪发生RN-突变的根本原因。AMPK-γ3复合物在糖酵解骨骼肌中对糖和脂肪的代谢有着独特的调节作用。本文系统综述了PRKAG3基因在骨骼肌中营养代谢调节作用和对肉质的影响及其基因表达的调控,为从分子水平深入开展肉质的研究提供参考。     

PRKAG3基因在不同品种猪不同生长阶段骨骼肌中的表达差异及其表达量与肉质的关系

本试验中旨在比较PRKAG3基因在不同品种猪不同生长阶段骨骼肌中的表达差异,并探讨PRKAG3基因与肉质的关系。挑选15 kg左右的汉普夏阉公猪17头和长撒阉公猪16头,饲喂相同饲粮,当体重分别达到20和50 kg时,2个品种的猪分别屠宰5头,体重达到100 kg时分别屠宰7和6头。各生长阶段屠宰后均测定骨骼肌pH、肌糖原含量以及PRKAG3基因表达量,且在100 kg阶段屠宰后同时测定肉质性状。结果表明:1)在不同生长阶段长撒猪骨骼肌中PRKAG3基因的表达量均高于汉普夏猪,特别是在100 kg阶段,长撒猪骨骼肌中PRKAG3基因的表达量是汉普夏猪的6.81倍(P0.05)。长撒猪与汉普夏猪骨骼肌中PRKAG3基因的表达量均随体重的增加而增加,但汉普夏猪不同生长阶段PRKAG3基因的表达量差异不显著(P0.05),而长撒猪PRKAG3基因的表达量在100 kg阶段时显著高于20和50 kg阶段时(P0.05)。2)汉普夏猪和长撒猪的肉质存在差异。汉普夏猪的滴水损失和失水率显著高于长撒猪(P0.05),而熟肉率、黄度(b)值极显著低于长撒猪(P0.01),剪切力和pH2(屠宰后24 h的pH)显著低于长撒猪(P0.05)。与长撒猪相比,汉普夏猪具有较高的肌糖原含量(P0.05)。3)猪骨骼肌中PRKAG3基因的表达量与肉质的相关性存在品种效应。汉普夏猪骨骼肌中PRKAG3基因表达量与滴水损失呈正相关,与熟肉率呈负相关,与pH2呈显著负相关(P0.05)。长撒猪骨骼肌中PRKAG3基因表达量与滴水损失和失水率呈正相关,与pH2呈负相关。上述结果表明,猪骨骼肌中PRKAG3基因具有品种和生长阶段表达差异;猪骨骼肌中PRKAG3基因的表达量与肉质性状相关,特别是与pH2,二者呈显著负相关。     

Association of SNPs in exon 2 of the MHC B-F gene with immune traits in two distinct chicken populations: Chinese Beijing-You and White Leghorn

Abstract

Antibody titers raised for vaccinations against avian influenza (AI) and Newcastle disease (ND) were higher in Chinese Beijing-You (BJY) than in White Leghorn (WL) (P<0.001), but there was no breed difference in titers for sheep red blood cells (SRBC). Genotyping by PCR-SSCP identified seven haplotypes in WL and 17 in BJY. After sequencing PCR products (35 and 85, respectively), 43 (WL) and 47 (BJY) single nucleotide polymorphisms (SNPs) were found in the 264 bp of exon 2. In WL chickens, significant associations were found with antibody responses to AI (two SNPs), ND (six SNPs), and SRBC (one SNP), while in BJY there was association with responses to ND (two SNPs) and SRBC (two SNPs), but none with AI. These results indicate that the genomic region bearing exon 2 of the major histocompatibility complex B-F gene has significant effects on antibody responses to SRBC and vaccination against AI and ND. Different SNPs affected antibody titers for each of the antigens and they differed between these very distinct breeds.     

Allelic frequencies of PRKAG3 in several pig breeds and its technological consequences on a Duroc × Landrace‐Large White cross

The allelic frequencies of PRKAG3 gene (the RN gene) have been investigated in several pig breeds. R200Q mutation appear only in Hampshire pigs, whereas V199I mutation is most abundant in Iberian, Porco Celta or Bizaro, and less in breeds selected for muscularity as Duroc, Landrace and Pietrain. A thorough study of phenotypic effects of V1991 has been performed in a Duroc × Landrace‐Large White cross. 199I homozygous pigs show increased pH₂₄ values in ham homogenates and loin (0.14 and 0.16 pH units, respectively) compared to 199V homozygous ones. Meat of 199I homozygous pigs exudates 42.6% less fluid and is darker (2.46 ‘L’‐value units). 199I homozygous pigs are fatter (4.2 mm more backfat thickness) and contain less muscle mass in ham (1.0 percentage points) and shoulder (2.7 percentage points), than 199V homozygous ones. 199I homozygous pigs contain 7.3% less protein in the belly and 8.5% more fat in shoulder muscle mass than 199V homozygous pigs. 199I homozygous pigs have also superior functional properties: better gelling (22.8% larger G′ value) and emulsion capacities (14 percentage points less of total exuded fluid), and higher curing yield in the belly (6 percentage points more). These data support the adipogenic character of the V199I mutation. The advantages and disadvantages of selecting any of the two PRKAG3 alleles for position 199 are discussed.     

PRKAG3在猪组织器官中的表达差异及与胴体品质关系研究

PRKAG3是编码一磷酸腺苷激活蛋白激酶(AMP-activated protein kinase,AMPK)γ3亚基的基因,是近年来确定的一个影响猪肉pH值、肉色以及系水力的主效基因,其突变R200Q与过量肌糖原含量有关,被认为是汉普夏猪发生RN-突变的根本原因。本研究以-βactin作为内参,用半定量RT-PCR方法测定雅南猪和杜×长×大三元杂交猪(DLY猪)PRKAG3基因在心脏、肝脏、肾和骨骼肌中表达差异,并测定猪胴体品质。结果表明,PRKAG3主要在骨骼肌中表达,在心脏中有少量表达,而在肝脏和肾中未见表达,在不同品种的骨骼肌中,雅南猪PRKAG3基因的相对表达量高于DLY猪(P=0.078)。相关分析结果表明:骨骼肌中PRKAG3基因的表达量与屠宰率、眼肌面积和背膘厚关系不大,而与肌内脂肪含量成正相关(r=0.832)。     

延边黄牛PRKAG3基因第10外显子单核苷酸多态性及其与肉质性状的相关分析

采用PCR-SSCP方法对120头延边黄牛PRKAG3基因的多态性进行检测,结果发现第10外显子存在C4738T突变,产生3种基因型。不同基因型与肉质性状均值差异显著性检验结果发现,基因型CT与a2*4、b2*4、油酸存在一定的负相关(P0.05);与嫩度、谷氨酸、精氨酸、丙氨酸、缬氨酸和脯氨酸有一定的正相关(P0.05);基因型CC与天冬氨酸存在一定的正相关(P0.05)。     

New alleles of chicken CD8 alpha and CD3d found in Chinese native and western breeds

Chinese native chicken breeds provide useful resources for the study of genetic diversity. In this study, the alleles of CD8 alpha and CD3d cDNA from Chinese native and introduced western breeds of chicken were analyzed at the sequence level. Six alleles were found, due to 13 amino acid replacements in the extracellular domain of the CD8 alpha sequence. There were four alleles detected in the Chinese strains, and alleles 5 and 6 were identified for the first time. Allele 6 was shared by Langshan, Beijing Fatty and Recessive White Feather chickens. Allele 2, found in the Bigbone strain, was the same as that found in the Leghorn strain H.B15.H7, and allele 3 in the Xianju breed was also the same as in the Leghorn strain H.B15.H12. Two Leghorn lines (RPL line 7 and AY519197) and the Camellia possessed an allele (alleles 1, 4 and 5), respectively, that was not found in the other lines. Nine out of 13 amino acid replacements were situated in the putative major histocompatibility complex (MHC) class I binding CDR1 (positions 30, 33 and 34), CDR2 (positions 58, 62, 63 and 65) and CDR3 (positions 90 and 106). Except for the Xianju breed, the CD8 alpha cDNA of Chinese native chicken breeds shared high homology. Two alleles were found in CD3d. Three additional nucleotides were found at positions 64, 65 and 66 in the newly discovered allele 2. This led to a difference of four amino acids (at residues 22, 23, 24 and 25) in the extracellular domain of CD3d cDNA from the Gushi, Recessive White Feather and ISA chickens compared with these of the White Leghorn and T11.15 (NM_205512). Five hybridoma clones (1C9, 1H5, 4B11, 6G5 and 13C5) against chicken CD8 alpha were generated by DNA immunization. Two monoclonal antibodies (mAbs), 6G5 and 4B11, showed reactivity to the splenocytes from five Chinese native chicken breeds, the Recessive White Feather chicken and the Leghorn (AY519197), while mAbs 1C9, 1H5 and 13C5 showed no reaction with these breeds.     

Exploring polymorphisms and their effects on reproductive traits of the INHA and INHβA genes in three goat breeds

In this study, we report the analysis of INHA and INHβA gene polymorphisms in 786 goats of three breeds: Xinong Saanen (SN), Guanzhong (GZ) and Boer (BG). We identified three new allelic variants: P1–C80G and/126G (GenBank accession no. HQ202573) in the three goat breeds and P2–C936T (GenBank accession no. HQ202572) in SN and GZ goat breeds. At P1 locus, AA, AB and BB genotypes were found in the three goat breeds. At P2 locus, CC and CT genotypes were found in SN and GZ goat breeds. After comparing genotype distribution within the three goat breeds, BG had conspicuous differences from SN and GZ (P < 0.001) at P2 locus. The SNP locus was in Hardy–Weinberg disequilibrium at P1 locus in the three goat breeds (P < 0.05). At P2 locus, the SNP locus was in Hardy–Weinberg disequilibrium in SN and GZ goat breeds (P < 0.05). Association of polymorphisms with litter size was done at P1 locus in the three goat breeds. The result showed that AA genotype had remarkable litter size at P1 locus in the three goat breeds (P < 0.05). Therefore, these results suggest that INHA gene is a strong candidate gene that affects litter size in goats.     

Similar developmental fluctuations of hepato-renal xanthine oxidoreductase gene expression and xanthine oxidase activity in layer and broiler chicken embryos

1. Xanthine oxidase (XO) has many physiological functions associated with the synthesis of both antioxidant (uric acid: UA) and numerous oxidants (e.g. H₂O₂), which makes it an important regulator of the cellular redox potential involving organogenesis. The ontogenetic study of hepatic and renal XO makes a better understanding of the putative role of this enzyme in the development of these tissues.

2. Developmental changes of gene expression of xanthine oxidoreductase (XOR), XO activity and UA content of liver and kidney tissues in both broiler and layer chicken embryos were examined during incubation d 14–21.

3. In both strains, hepatic XOR gene expression peaked on d 21 while renal XOR gene expression did not change.

4. The XO activity was higher in kidney than liver in both strains. Hepatic XO activity of both strains peaked on d 18 and thereafter was decreased on d 21. Renal XO activity peaked on d 18 and from then on did not show any significant changes until d 21 in both strains.

5. The UA content was higher in kidney vs. liver in both strains. The hepatic and renal UA values of the both strains increased significantly from d 14 to d 21.

6. The present results showed dissimilar behaviour of XOR gene expression, XO activity and UA content of liver and kidney tissues in both broiler and layer chicken embryos.     

Association of three beta-defensin gene (AvBD4, AvBD5, AvBD14) polymorphisms with carrier-state susceptibility to salmonella in chickens

ABSTRACT

1. Chicken salmonellosis is a common zoonotic infectious disease transmitted both vertically and horizontally. Avian beta-defensins (gallinacins) play an important role in the innate defence of the host and provide broad-spectrum immunity against multiple pathogens.

2. To detect the relationship between immune genes and salmonella carrier status and susceptibility to salmonellosis in chickens, polymorphisms with carrier-state susceptibility to salmonella and, hence, developing salmonellosis, were investigated in three avian beta-defensin genes (AvBD4, AvBD5, and AvBD14) in a Chinese local chicken breed, based on a case-control study.

3. Fifteen, twenty and nineteen SNPs were found in AvBD4, AvBD5 and AvBD14, respectively. Among the 54 total SNPs, four resulted in non-synonymous substitution of amino acid changes. Five SNPs in AvBD5 and four SNPs in AvBD14 were significantly associated with salmonellosis susceptibility (P < 0.05). Using the PHASE program, thirteen, ten and twelve major haplotypes were constructed in AvBD4, AvBD5 and AvBD14. Logistic regression analysis revealed that five haplotypes in AvBD5 and six haplotypes in AvBD14 were significantly associated with salmonellosis susceptibility, but no significant haplotype in AvBD4 was detected. A total of six strongly susceptible haplotypes with odds ratio (OR) values greater than 2.0 and four strongly resistant haplotypes with OR value less than 0.5 were revealed in the three genes examined.

4. These results suggested that the AvBD5 and AvBD14 genes may play an important role in the susceptibility to salmonellosis in chickens.     

Effect of 3,3',5-triiodothyronine and 3,5-diiodothyronine on progesterone production, cAMP synthesis, and mRNA expression of STAR, CYP11A1, and HSD3B genes in granulosa layer of chicken preovulatory follicles

In vitro studies were performed to assess whether stimulatory effects of triiodothyronine (T₃) on progesterone (P₄) production in a granulosa layer (GL) of chicken preovulatory follicles are associated with 3′,5′-cyclic adenosine monophosphate (cAMP) synthesis and mRNA expression of STAR protein, CYP11A1, and HSD3B. Effects of 3,5-diiodothyronine (3,5-T₂) on steroidogenic function in these follicles were also investigated. The GL of F3 to F1 follicles was incubated in medium supplemented with T₃ or 3,5-T₂, LH, or forskolin (F), and a combination of each iodothyronine with LH or F. Levels of P₄ and cAMP in culture media were determined by RIA. Expression of genes involved in P₄ synthesis (ie, STAR protein, CYP11A1, and HSD3B) in the GL of F3 to F1 follicles incubated in medium with T₃ or 3,5-T₂ and their combination with LH was performed by real-time PCR. Triiodothyronine increased basal and LH- and F-stimulated P₄ secretion by preovulatory follicles. The 3,5-T₂ elevated P₄ synthesis by F3, had no effect on F2 follicles, and diminished P₄ production by the GL of F1 follicles. It had no effect on LH-stimulated P₄ production; however, it augmented F-stimulated P₄ production by F2 and F1 follicles. Although T₃ did not affect basal and F-stimulated cAMP synthesis by the GL of preovulatory follicles, it increased LH-stimulated synthesis of this nucleotide. However, 3,5-T₂ elevated F-stimulated cAMP synthesis in F3 and F2 follicles; it did not change basal and LH-stimulated cAMP production. Triiodothyronine decreased basal STAR and CYP11A1 mRNAs in F3 follicles, increased them in F1 follicles, and elevated HSD3B mRNA levels in F1 follicles. Triiodothyronine augmented LH-stimulated STAR, CYP11A1, and HSD3B mRNA levels in F2 and CYP11A1 in F1 follicles. However, T₃ decreased LH-stimulated STAR and HSD3B mRNA levels in F1 follicles. The 3,5-T₂ did not affect basal STAR and CYP11A1 mRNA expression in all investigated follicles; however, it decreased LH-stimulated STAR expression in F2 and F1 ones. The effects of 3,5-T₂ caused elevated basal but diminished LH-stimulated HSD3B mRNA levels. In conclusion, data indicate that both iodothyronines are involved in P₄ production in the GL of chicken preovulatory follicles acting alone and additively with LH. Effects of iodothyronines depend on follicle maturation and are associated with modulation of cAMP synthesis and STAR, CYP11A1, and HSD3B mRNA expression. We suggest that iodothyronines participate in maturation and ovulation of chicken follicles.     

Detection of novel single-nucleotide polymorphisms (SNPs) in the CYP21 gene and association analysis of two SNPs for CYP21 and ESR2 with litter size in a commercial sow population

Altogether 129 F₁ sows from a commercial sow farm with at least four litters were genotyped for the oestrogen receptor 2 gene (ESR2) and cytochrome P450 hydroxylase 21 gene (CYP21) and investigated for associations on the litter‐size parameters: total number born and number born alive. Five novel polymorphisms were found in the 3′‐untranslated region for the CYP21 gene. Genotype and allele frequencies for the CYP21 (position 3462G > A) single‐nucleotide polymorphism (SNP) were 0.434 (GG), 0.504 (AG), 0.062 (AA) and 0.69 (G):0.31 (A), respectively. No association was found between this polymorphism and litter‐size parameters. For the ESR2 gene, the SNP in exon 5 associated with an amino acid substitution MET (allele A) > VAL (allele G) was investigated. Only two genotypes were found leading to allele frequencies of 0.34 (A):0.66 (G). Only number born alive piglets were significantly increased for the AG genotype (p = 0.034) with 11.64 piglets per sow and litter in comparison with the GG genotype, leading to only 10.96 piglets per sow and litter. From these data, it can be concluded that the investigated SNP of the ESR2 gene is associated with the number of liveborn piglets in the commercial population considered, and hence could be useful in selection for litter size. Therefore, this gene should be investigated in additional populations.     

中国荷斯坦奶牛铁蛋白基因3'端克隆及序列分析

本研究旨在克隆和分析中国荷斯坦奶牛的铁蛋白(FTH)基因,为该基因的功能研究和有效利用提供理论依据.运用cDNA末端快速扩增(RACE)技术从公犊肝脏组织中扩增出FTH基因的3'端cDNA序列,并利用生物信息学方法对该基因的氨基酸序列进行分析.结果表明,FTH基因3'端cDNA全长为489 bp,编码81个氨基酸;经生物信息学分析表明,该基因编码的蛋白无N端信号肽及跨膜区,相对分子质量为9.1253 kD,理论等电点为5.19,脂肪系数为85.56,总亲水性为-0.342,其一级结构中存在4个功能结构域;二级结构元件以α-螺旋和β-旋转为主;同源序列分析表明,中国荷斯坦奶牛与原牛FTH氨基酸的相似性最高(99％);系统进化树显示,在该基因座位上中国荷斯坦奶牛与所测物种的亲缘关系均较远.本研究有助于揭示FTH家族基因的进化历史,为对其进行深入的功能分析和利用研究提供参考.     

STAT5b基因的遗传多态性及其与京海黄鸡体重和繁殖性状的关联分析

以京海黄鸡、AA鸡、尤溪麻鸡、边鸡等4个鸡品种为研究对象,采用PCR-SSCP方法检测信号转导和转录激活子5b(STAT5b)第7外显子和部分内含子7的多态性,并分析其对京海黄鸡体重和繁殖性状的遗传效应。结果显示,在STAT5b第7内含子区域检测到1处突变(8 066bp C→T)。在4个鸡品种中均检测到AA、AB、BB 3种基因型,χ2检验结果表明,除京海黄鸡外其余3个品种群体在该座位均达到Hardy-Weinberg平衡(P>0.05)。最小二乘分析表明,京海黄鸡3种基因型个体的出生重和开产日龄存在显著差异(P<0.05)。     

Identification and characterization of polymorphisms within the 5' flanking region, first exon and part of first intron of bovine GH gene

The aim of the present study was to identify and characterize polymorphisms within the 5′ flanking region, first exon and part of first intron of the bovine growth hormone gene among different beef cattle breeds: Nelore (n = 25), Simmental (n = 39), Simbrasil (n = 24), Simmental × Nelore (n = 30), Canchim × Nelore (n = 30) and Angus × Nelore (n = 30). Two DNA fragments (GH1, 464 bp and GH2, 453 bp) were amplified by polymerase chain reaction and then used for polymorphism identification by SSCP. Within the GH1 fragment, five polymorphisms were identified, corresponding to three different alleles: GH1.1, GH1.2 and GH1.3 (GenBank: AY662648 , AY662649 and AY662650 , respectively). These allele sequences were aligned and compared with bovine GH gene nucleotide sequence (GenBank: M57764 and AF118837 ), resulting in the identification of five insertion/deletions (INDELs) and five single nucleotide polymorphisms (SNPs). In the GH2 fragment two alleles were identified, GH2.1 and GH2.2 (GenBank: AY662651 and AY662652 , respectively). The allele sequences were compared with GenBank sequences ( M57764 , AF007750 and AH009106 ) and three INDELs and four SNPs were identified. In conclusion, we were able to identify six new polymorphisms of the bovine GH gene (one INDEL and five SNPs), which can be used as molecular markers in genetic studies.