热应激下鸡肝脏SFRP1和PDK4基因表达水平研究

为研究热应激条件下,鸡SFRP1和PDK4两个与代谢相关的基因在鸡肝脏组织中的差异表达,选取12只60日龄体重相近的黄羽肉母鸡,分为正常对照组(CL)、热应激反应组(HS)和热应激反应后的降温组(HF)3个处理组,并对这两个基因在三组肝脏组织中的表达水平进行荧光定量表达分析。结果表明,在热应激状态下,SFRP1和PDK4基因在肝脏中的表达均上调,且PDK4在对照组和热应激组间差异显著(P<0.05)。本研究结果还表明,当热应激反应后温度重新降至(25±1)℃时,SFRP1和PDK4在肝脏中的相对表达量均有不同程度的恢复,进一步说明这两个基因在热应激中发挥的作用。本研究为从分子机制分析热应激提供了相应的参考。     

卢氏绿壳蛋鸡壳色性状的AY493302标记

卢氏鸡是我国唯一片羽型的可产绿壳蛋的地方鸡种,壳色有绿壳和褐壳2种颜色。本研究采集131只卢氏绿壳蛋鸡,45只卢氏褐壳蛋鸡及18只地方白壳蛋鸡的血液,提取基因组DNA,用微卫星AY493302标记探讨了这3种壳色的基因型差异。结果表明,微卫星AY493302在卢氏绿壳蛋鸡、卢氏褐壳蛋鸡以及地方白壳蛋鸡之间基因型分布差异极显著,基因型AA（225bp/225bp）在卢氏鸡中的频率为0.396 9,且只在卢氏绿壳蛋鸡中出现。其A基因频率在卢氏绿壳蛋鸡、卢氏褐壳蛋鸡和地方白壳蛋鸡中分别为0.698 5,0.500 0,0.500 0。进一步进行测序发现,卢氏绿壳蛋鸡AA基因型在19bp处碱基和217bp处碱基均为G,但卢氏绿壳蛋鸡AB基因型在19bp处碱基却为C,在217bp处碱基与绿壳蛋鸡AA基因型同为G。卢氏褐壳蛋鸡AB基因型和地方白壳蛋鸡的AB基因型则表现在19bp处碱基为C,217bp处碱基为T。AA基因型的序列与NCBI数据库中红色原鸡比较,达99%同源的序列有V00425.1、J02015.1和NW_003763484.1,均位于鸡的1号染色体上。该微卫星标记的群体杂合度、群体多态信息含量在卢氏绿壳蛋鸡中都低于卢氏褐壳蛋鸡和地方白壳蛋鸡。尽管在所研究的卢氏绿壳群体中只有39.7%的个体表现为纯合AA基因型,但卢氏褐壳蛋鸡和地方白壳蛋鸡中没有该基因型,故而认为微卫星AY493302的纯合AA基因型可以作为对卢氏绿壳蛋鸡进行分子标记辅助选择的基因型。     

Nutritional significance of amino acids,vitamins and minerals as nutraceuticals in poultry production and health – a comprehensive review

Nutraceuticals have gained immense importance in poultry science recently considering the nutritional and beneficial health effects of their constituents. Besides providing nutritional requirements to birds, nutraceuticals have beneficial pharmacological effects, for example, they help in establishing normal physiological health status, prevent diseases and thereby improve production performance. Nutraceuticals include amino acids, vitamins, minerals, enzymes, etc. which are important for preventing oxidative stress, regulating the immune response and maintaining normal physiological, biochemical and homeostatic mechanisms. Nutraceuticals help in supplying nutrients in balanced amounts for supporting the optimal growth performance in modern poultry flocks, and as a dietary supplement can reduce the use of antibiotics. The application of antibiotic growth enhancers in poultry leads to the propagation of antibiotic-resistant microbes and drug residues; therefore, they have been restricted in many countries. Thus, there is a demand for natural feed additives that lead to the same growth enhancement without affecting the health. Nutraceuticals substances have an essential role in the development of the animals’ normal physiological functions and in protecting them against infectious diseases. In this review, the uses of amino acids, vitamins and minerals as well as their mode of action in growth promotion and elevation of immune system are discussed.     

Assessment of age-related morphological changes in the testes of post-hatch light ecotype Nigerian indigenous chicken

The age-related morphological changes of the testes in light ecotype Nigerian indigenous chicken were evaluated in this study using gross anatomical, histological and histomorphometric techniques. The results showed that the testes of 3- to 9-month-old birds were light pink while testes of sexually mature chicken were creamy white in colour. The left and right testicular weight, length, diameter, circumference and the organosomatic indices increased significantly (p < .05) with increasing age across the groups. Although the mean tubular diameter and epithelial height of the left and right seminiferous tubules increased significantly (p < .05) with age, the tubular diameter, epithelial height and luminal diameter did not vary significantly (p > .05) between the left and right testes of all the groups. The one-cell layer thick germinal epithelium of the left testes at 3 to 6 months old showed islands of cell proliferation that contained spermatogonia and spermatocytes. At 6 to 9 months, the left testes exhibited numerous early spermatids with occasional occurrence of late stage spermatids while the right testes showed scanty early stage spermatids. At 12 to 18 months, the germinal epithelia of both left and right testes were characterized by the presence of Sertoli cells, spermatogonia, primary spermatocytes, numerous early and late stage spermatids as well as spermatozoa. In conclusion, the morphological features highlighted in the present study show that at pre-pubertal periods, the left testes may develop faster than the right testes. However, both left and right testes may participate actively in the production of spermatozoa during the post-pubertal life.     

鸡繁殖性能近交衰退相关CpG岛差异甲基化基因的筛选

鸡繁殖性能近交衰退是地方鸡遗传资源活体保种过程中面临的重要问题之一,本研究旨在探讨全基因组CpG岛（CpG island,CGI）区DNA甲基化在鸡繁殖性能近交衰退中的作用。分别从狼山鸡高近交组和低近交组中各选取健康母鸡3只,即试验分2个组,每组3个重复,然后采用全基因组重亚硫酸盐测序（WGBS）技术,检测分析两组个体性腺轴组织（包括卵巢和下丘脑）全基因组DNA甲基化差异,筛选差异甲基化区域（DMRs）,并对CpG岛区差异甲基化基因进行功能注释和富集分析。结果表明,狼山鸡高近交组和低近交组比较,其卵巢和下丘脑基因组整体甲基化水平均不存在显著差异（P>0.05）;高、低近交组间差异甲基化区域检测发现,下丘脑和卵巢中分别检测到5 948和4 593个差异甲基化区域,其中1 798和995个差异甲基化区域位于基因组CpG岛区,分别注释到1 020和552个基因;下丘脑中,这些CpG岛区差异甲基化基因显著富集在信号转导、神经系统发育、生殖系统发育和卵母细胞成熟调控等繁殖相关的GO条目,以及转化生长因子β信号通路、乙型肝炎、脂肪酸代谢、胰岛素信号通路等19条KEGG信号通路（P<0.05）;卵巢中,CpG岛区差异甲基化基因显著富集于12条信号通路（P<0.05）,包括慢性骨髓白血病、流感A、精氨酸和脯氨酸代谢、粘着连接等,一些与卵子发育和性激素分泌相关的信号通路也被富集到,如黄体酮介导的卵母细胞成熟、卵母细胞减数分裂、GnRH信号通路、雌激素信号通路等,其中包含CDC27、ADCY8、AKT3等10个差异甲基化基因。因此,本研究在狼山鸡高、低近交组间检测到了大量差异甲基化区域,并发现大量差异甲基化基因与繁殖性状相关,推测这些基因CpG岛区DNA甲基化可能在狼山鸡繁殖性能近交衰退调控中发挥重要作用,研究结果为进一步深入探索鸡繁殖性能近交衰退调控机制奠定了基础,为物种资源保护和家禽育种工作提供了理论参考依据。     

1例肉鸡肝恶性黑色素瘤的病理学诊断

本研究旨在对临床上发现的1例肉鸡肝黑色素瘤进行病理学诊断。1羽25日龄AA肉鸡精神沉郁、极度消瘦,剖检发现其整个肝呈黑色。HE染色结果显示,病变肝内遍布大小不一、细胞核深染、细胞质内有大量黑色素颗粒沉积的异常细胞团。黑色素瘤诊断标志蛋白SOX 10和HMB-45抗体免疫组化染色均呈阳性。结合病理学检测和免疫组化结果,判定该病鸡肝患有恶性黑色素瘤。试验对病变肝进行胆色素染色和普鲁士蓝染色,结果均呈阴性,提示病变肝并非由胆红素或含铁血黄素淤积所致。综上表明,此例病鸡患有严重的肝恶性黑色素瘤,试验结果对禽类黑色素瘤的诊断与研究有参考价值。     

参考群筛选方法及规模对基因型填充准确性的影响

为探究基于A矩阵期望遗传关系最大化（maximizing the expected genetic relationship for matrix A,RELA）、基于A矩阵目标群体遗传方差最小化（minimized the target population genetic variance for matrix A,MCA）、平均亲缘关系最大化（the highest mean kinship coefficients,KIN）、随机选择（random selection,RAN）、共同祖先筛选（common ancestor,CA）等不同参考群筛选方法及参考群规模对基因型填充准确性的影响。本研究使用矮小型黄羽肉鸡作为试验群体,采用鸡600K SNP芯片（Affymetrix Axion HD genotyping array）进行基因分型,测定435羽子代公鸡45、56、70、84、91日龄体重。利用Beagle软件将低密度SNP芯片填充为高密度SNP芯片数据,比较不同参考群筛选方法、参考群规模对基因型填充准确性的影响,以及填充芯片基因组预测准确性。结果表明,使用Beagle 4.0结合系谱信息进行填充效果最佳,其次为Beagle 4.0,而Beagle 5.1填充效果最差。使用MCA方法筛选参考群进行基因型填充准确性最高,使用RAN方法筛选参考群进行基因型填充准确性最低,MCA、RELA、CA 3种方法基因型填充准确性差别较小。相比其他方法,使用MCA方法筛选个体作为参考群将低密度SNP芯片填充至高密度SNP芯片进行基因组选择的预测准确性较高,与真实高密度SNP芯片的基因组预测准确性相差甚微。随着参考群规模增大,基因型填充准确性也随之增加,但增速逐渐下降,最后趋于平缓。综上所述,可以通过参考群筛选方法构建参考群以及控制参考群规模,以保证基因型填充和基因组预测准确性并节省成本,本研究为基因型填充在畜禽遗传育种中的应用提供技术参考。     

Egg production,fertility, hatchability and immune responses of some chicken strains under high ambient temperatures

This experiment was designed to investigate and compare the egg production, fertility, hatchability and immune responses of some local developed Egyptian chicken strains under high ambient temperatures. A total of 108 (26 weeks old) laying hens of Matrouh, Silver Montazah, Mandarah and Inshas (9 hens × 3 replicates × 4 strains) were used to evaluate the impact of thermal stress (24-34ºC) during summer season (June, July and August) on egg production, fertility, hatchability and immune responses. The obtained results revealed that final body weight (FBW/g) and body weight change (BWC/g) among different chicken strains were not affected. The daily feed consumption (FC/g) and feed conversion ratio (FCR) for Silver Montazah and Inshas strains were significantly (p ≤ .05) higher than those of Matrouh and Mandarah strains. The mean egg production (EP/%) and egg mass (EM) for Silver Montazah and Inshas strains were significantly (p ≤ .05) higher than those of Matrouh and Mandarah strains. The highest percentages of hatchability of total eggs (HTE) and fertile eggs (HFE) were recorded in Inshas chickens, while the lowest value was recorded in Mandarah chickens. Also, the highest embryonic mortalities (p ≤ .05) and lowest chick weight at hatch (CWH/g) were recorded in Mandarah chickens compared with the other strains. The highest values for antibody titres against phytohemagglutinin-P (PHA-P) were recorded in Mandarah chickens, while the least values were recorded in Inshas chickens.     

0~4周龄云岭红肉鸡赖氨酸适宜需要量的研究

试验通过分析饲粮不同赖氨酸水平对云岭红肉鸡生长性能、氮沉积率及血液生化指标的影响,确定云岭红肉鸡饲粮赖氨酸的适宜需要量。采用单因素试验设计,将180只健康状况良好、体重相近的1日龄云岭红肉鸡随机分为5个处理组,每组3个重复,每个重复12只鸡。5组试验鸡饲喂的饲粮赖氨酸水平分别为0.85%、0.95%、1.05%、1.15%、1.25%。结果表明:赖氨酸水平对试验鸡的期末体重、全期增重、平均日增重及料重比有显著影响(P<0.05),当赖氨酸水平为1.15%时期末体重最大739.21 g,全期增重最大701.66 g,平均日增重最大24.19 g,料重比最低1.68。不同赖氨酸水平对丝氨酸、甘氨酸和组氨酸的表观代谢率影响显著(P<0.05)。对血清总氨基酸和血清尿素氮影响显著(P<0.05)。研究表明,0~4周龄云岭红肉鸡适宜赖氨酸水平最适量为1.15%。     

鸡FKBP5基因的克隆、生物信息学及组织表达谱分析

本研究旨在对鸡FK506结合蛋白5(FK506 binding protein 5,FKBP5)基因进行克隆和生物信息学分析,并检测其在鸡不同组织中的表达量。以鸡脾脏cDNA为模板,通过PCR方法扩增和克隆鸡FKBP5基因完整CDS区序列,并进行同源性比对及系统进化树构建;使用在线软件分析FKBP5蛋白的理化性质、疏水性、跨膜结构、信号肽、二级结构和三级结构;利用实时荧光定量PCR检测其在肢体内外翻畸形(valgus-varus deformity,VVD)组肉鸡和正常组肉鸡组织中的表达量,并绘制组织表达谱。结果显示,鸡FKBP5基因CDS区序列全长1350 bp,编码449个氨基酸;同源性比对和进化树分析表明,鸡FKBP5基因氨基酸序列与鹌鹑、鸭、壁虎、中华鳖、小鼠、人、猪、斑马鱼的同源性分别为98.4%、96.2%、85.8%、87.4%、81.1%、85.2%、86.1%和61.2%,与鹌鹑、鸭的亲缘关系最近,爬行类和哺乳类动物次之,与斑马鱼(鱼类)亲缘关系最远。鸡FKBP5蛋白分子质量为50.43 ku,理论等电点(pI)为5.94,半衰期为30 h,肽链N端为蛋氨酸(Met),不稳定系数为23.80,属于稳定蛋白;跨膜区和信号肽预测结果显示,FKBP5蛋白不属于跨膜蛋白和分泌型蛋白。结构域分析结果表明,该蛋白包括2个FKBP型肽基脯氨酰异构酶(FKBP-type peptidylprolyl isomerases)、3个四肽重复(TPR)序列,主要包含α-螺旋(46.77%)、延伸链(12.47%)和无规则卷曲(40.76%),且该蛋白与HSPA2、PPID、STIP1、HSP90AA1和HSP90AB1等互作蛋白具有很强的相关性。实时荧光定量PCR结果显示,FKBP5基因在鸡各组织中广泛表达,其中在软骨和法氏囊中表达量较高,在发病组肉鸡组织中的表达量均高于正常组肉鸡,且在心脏、腿肌、法氏囊、胸腺、软骨中表达量差异显著(P<0.05),在脾脏中表达量差异极显著(P<0.01)。本试验结果可为肉鸡骨骼疾病及腿部健康选育提供参考。