滴鼻免疫疫苗对SPF鸡气管功能影响的研究

为研究疫苗滴鼻免疫对鸡气管功能的影响，本研究对SPF鸡滴鼻免疫新城疫疫苗后气管纤毛摆动速率、气管形态结构和黏液分泌进行了研究。80只7日龄SPF鸡随机分为对照组和试验组，每组40只，试验组SPF鸡滴鼻免疫新城疫弱毒疫苗，对照组滴鼻生理盐水；于免疫后的第0、2、4和6天，每组每次随机剖杀10只鸡，应用红细胞示踪法测定气管纤毛摆动速率，采集气管制备石蜡组织切片进行HE和奥新蓝pH 2.5和pH 1.0染色。结果发现，疫苗免疫后的第0和2天试验组SPF鸡纤毛摆动速率较对照组升高，但差异不显著（P>0.05）；滴鼻免疫后SPF鸡气管结构完整，无明显损伤；滴鼻免疫后试验组SPF鸡气管酸性黏液分布容积在免疫后的第0和2天显著高于对照组（P<0.05），而后明显降低，于免疫后的第6天又有所升高，但差异不显著（P>0.05）；硫酸化黏液分布容积在免疫后的第0和2天显著升高（P<0.05），而后降低并与对照组相一致。结果表明，新城疫疫苗滴鼻免疫后前两天SPF鸡气管黏液分泌增多，而后恢复到正常水平，疫苗的免疫对气管形态结构和纤毛的摆动速率无明显影响。     

鸡无尾性状的分子机制研究进展

动物的尾巴具有改变方向、控制升降、调整速度、支撑身体、防御、攻击、保温、示警、逃生和捕食等作用。鸟类的尾巴在飞行过程中起到平衡身体的关键作用，但在进化过程中尾部却出现了缩短和融合现象，如中国的瓢鸡及智利的Araucana鸡。作者简述了近年来关于鸡无尾性状的解剖学研究成果，发现无尾鸡在胚胎发育HH19期（Hamburger和Humilton标准分期）就已经出现无尾现象，而在结构上无尾鸡缺乏尾脂腺、尾羽、镰羽、尾椎骨和尾综骨。同时作者分析了导致尾部体节停止发育的分子遗传机制，包括周期性表达基因的转录调控影响尾部延伸过程、后端化因子梯度诱导尾部形成、Hox基因影响体节特化过程及基因突变有可能导致的无尾现象等，从而提出了可能导致家禽无尾性状的胚胎发育时期的关键分子及信号通路，包括Irx1和Irx2基因、Notch信号通路、Wnt信号通路、Fgf信号通路、RA信号通路等。研究鸡无尾性状的分子机制不仅可深入了解脊椎动物胚胎发育中尾部发育机制，更有利于揭示鸟类在进化过程中发生的尾部缩短及融合的机理。     

鸡粪及其配施尿素对甜瓜香味物质、酶活性及基因表达的影响

为了探讨鸡粪以及鸡粪与尿素配施对薄皮甜瓜果实香气物质合成及关键酶的影响,以薄皮甜瓜"DX108"为试材,以尿素为氮源的处理作为对照(CK),研究等量氮素条件下,鸡粪及其与尿素配施处理对甜瓜果实香气物质以及相关酶活性变化的影响,并分析了乙醇脱氢酶(ADH)和醇酰基转移酶(AAT)基因的表达水平。结果表明:不同处理对薄皮甜瓜香气物质合成、相关酶活性以及关键酶基因表达的影响存在明显差异;尿素处理促进了甜瓜果实中C6和C9醇醛类化合物的合成,而鸡粪以及鸡粪与尿素配施提高了成熟果实中酯类物质含量和种类,尤其是乙酸酯类含量;鸡粪配施尿素处理提高了花后35 d甜瓜果实中非乙酸酯类物质含量,且是尿素处理花后35 d甜瓜果实中非乙酸酯类物质含量的3.1倍,还检测到了尿素处理果实中没有检测到的特征性酯类物质;鸡粪和鸡粪配施尿素处理提高了花后25～30 d果实中LOX酶、氨基转移酶、ADH酶以及AAT酶活性,降低了花后35 d果实中ADH酶活性,抑制了30 d后果实中CmADH1和ADH2的基因表达,而促进了CmAAT1和CmAAT3基因表达。由此可知,鸡粪以及鸡粪与尿素配施有可能是通过调节果实不同发育期香味物质合成途径中关键酶活性的协调变化以及关键酶基因表达,影响了果实香气物质的合成,尤其是酯类物质的合成。     

Establishment of Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry Determination Method of Amantadine in Chicken and Preparation of Positive Samples

An ultra performance liquid chromatography (UPLC)-tandem mass spectrometry analysis method was established in this experiment for determination of amantadine in chicken.1% trichloroacetic acid with methanol as extraction liquid, the ultrasonic assisted solvent extraction was used to extract amantadine in chicken, extract liquid was purified and enriched in MCX solid-phase extraction column.Using C18 chromatographic column as separation column, in the positive ion mode, we detected the samples by electrospray ionization tandem mass spectrometry.Mass spectrometry and chromatographic conditions, the type of extract liquid, ultrasonic extraction time, the types of solid-phase extraction column and eluent were optimized.Under optimal conditions, a good linearity (r=0.9998) in the ranges of 0.1 to 20.0 ng/mL was obtained, the recoveries were 89.7% to 101.4% in the additive levels of 5.0, 10.0 and 20.0 μg/kg, the limit of detection (LOD) of amantadine was 0.07 μg/kg, the limit of quantiation (LOQ) of amantadine was 0.23 μg/kg, and the relative standard deviations (RSD) were below 7.0%.This novel approach had high sensitivity and accuracy, and was successfully applied to rapid and high sensitive detection of amantadine residues in chicken.The positive samples of amantadine in chicken were successfully prepared with concentrations of 11.16 and 7.18 mg/kg, which could be used for the research of ELISA kits of amantadine.     

雏鸡中枢免疫器官内B淋巴细胞变化规律

为了研究雏鸡在正常发育过程中免疫器官内B淋巴细胞的变化规律,以不同日龄的SPF鸡为研究对象,用流式细胞术检测鸡脾脏、胸腺及法氏囊内IgA+、IgG+及IgM+B淋巴细胞含量的变化情况。结果显示,在脾脏中,4日龄时B淋巴细胞含量比较多,7日龄时B淋巴细胞含量下降,随后,7至21日龄细胞含量逐渐升高;胸腺内则是在4日龄时IgM+B淋巴细胞含量相对较高,而IgA+、IgG+B淋巴细胞几乎没有;在法氏囊中,IgA+、IgG+及IgM+B淋巴细胞含量在4至14日龄时逐渐升高,并且IgA+和IgG+细胞含量在21日龄时基本趋于稳定,但是IgM+B细胞含量则在21日龄时有所下降。试验结果表明,雏鸡出壳后各免疫器官B淋巴细胞始终以IgM+细胞含量最多。     

Study on Expression of Bcl-2 and PCNA Genes in Testicular Tissue of Chickens,Quails and Chicken-quail Hybrids

In order to investigate the molecular mechanism of stunted growth testis of chicken (♂) and quail (♀) hybrids. 84 testicular tissue samples of New Roman cocks, male Korean quails and chicken-quail hybrids at different developmental stages were collected,the mRNA expression of Bcl-2 and PCNA genes in testicular tissue of cocks, quails and chicken-quail hybrids at different growth stages were detected using Real-time PCR. The results showed that the Bcl-2 and PCNA genes mRNA expression patterns in testes of chickens and quails at different growth stages were similar.The Bcl-2 gene was fluctuated in a whole, and decreased to the lowest value, then recovered rapidly and remained at a high level. The PCNA gene had a very significant peak value.Compared with the chicken and quail,the expression of Bcl-2 and PCNA genes mRNA in hybrids had no obvious change, which indicated that the pattern of the Bcl-2 gene regulation apoptosis in testis was different from chicken and quail at the hybrid testicular development process, and there was no obvious cell appreciation process.It was suggested that the molecular influencing factors of testicular dysplasia of chicken and quail hybrids were related to the abnormal expression of Bcl-2 and PCNA genes mRNA in testes.The results provided an important reference for the further study of the mechanism of testicular dysplasia between chicken and quail interspecific hybrids.     

禽呼肠病毒感染鸡胚成纤维细胞后Toll样受体mRNA转录水平的动态变化

为分析禽呼肠病毒(ARV)标准毒株S1133株感染鸡胚成纤维细胞(CEF)后对相关鸡Toll样受体(ChTLRs)mRNA转录水平的影响作用,利用实时荧光定量PCR,测定和分析ARV-S1133感染CEF后ARV结构蛋白σC和ChTLRs的mRNA转录水平变化情况。结果表明,ARV-S1133感染CEF 10h后,ARVσC蛋白的mRNA相对表达量开始迅速上升,在48h达到峰值;同时,感染CEF中的ChTLR3、ChTLR5、ChTLR7、ChTLR15和ChTLR21mRNA表达量发生显著变化,在感染72h内各个受体的mRNA表达量呈波浪式变化。5个不同滴度的ARV感染CEF 24 h后,ChTLR3、ChTLR5、ChTLR7、ChTLR15、ChTLR21mRNA转录水平与病毒滴度均呈正线性相关。上述结果表明,ARV感染后可诱导CEF ChTLR3、ChTLR5、ChTLR7、ChTLR15、ChTLR21的mRNA转录水平发生变化,可能与禽呼肠病毒的复制和致病机制相关。     

Effect of Different Rearing Conditions of Indoor and Outdoor in Winter and Summer on Growth Performance and Immune Function of Heijiao MA Chicken

This experiment was conducted to study the effect of different rearing conditions of indoor and outdoor in winter and summer on growth performance and immune function of Heijiao MA chicken.In the winter and summer, 180 Heijiao MA chicken with the age of 35-day-old were chosen and randomly divided into two groups with three replicates per group and 30 chickens per replicate.The Heijiao MA chicken were reared at outdoor and indoor, respectively, and at 49, 63, 77, 91 days the immune organ indexes, serum antibody titers and growth performance were measured.The results showed that:①Whether it was winter or summer, the body weight of Heijiao MA chicken rearing at indoor were higher than that of outdoor, and the difference was significant at experimental stages in winter (P<0.05).There was significant difference only at 77 days old of Heijiao MA chicken in summer (P<0.05), the rest were not significant (P>0.05);②The ADG of outdoor group in summer was higher than that in winter, while the F/G of outdoor group in summer were significantly lower than that in winter (P<0.05).The ADFI and ADG of indoor group were not significantly different between winter and summer (P>0.05).③On immune function, immune organ indexes and ND antibody titer of Heijiao MA chicken of indoor group in winter were higher than outdoor group, while part of immune organ indexes, AI (H5) and ND antibody titers of outdoor group in summer were higher than that of indoor group;And ND and AI (H5) antibody titers in winter were higher than summer.In conclusion, The climate and season were major factors to influence the growth performance and immune function of Heijiao MA chicken.     

Analysis on Target Genes with Mutation in microRNA of Blue-shelled Chicken and White Leghorn

The study was conducted to explore the potential different characters between Blue-shelled chicken and White leghorn.Global genome microRNA was combined the identified microRNA with complementary lab-predicted microRNA.Then the two breed chicken's SNP data got by GGRS were mapped to the microRNA and focused on SNP that deliberately located in mature-microRNA.Bioinformatics method was adopted for target prediction on microRNA which had SNPs.By further gene enrichment analysis,the study found these genes enriched in 22 GO terms,10 KEGG pathways,and 3 IPA important networks.And they enriched in traits which associated with growth,such as mTOR signaling pathways,Wnt signaling pathways,growth hormone receptor networks and insulin-like growth factor Ⅰ receptor networks.And they also enriched in some laying traits,such as oocyte meiotic signaling pathways and progesterone mature oocytes signaling pathways.The methods and the results might provide references for further studies.     

饲粮代谢能水平对14～17周龄不同性别北京油鸡育肥期生长性能、屠宰性能、肉品质及血清生化指标的影响

本试验旨在研究饲粮代谢能水平对14～17周龄不同性别北京油鸡育肥期生长性能、屠宰性能、肉品质及血清生化指标的影响，为确定北京油鸡适宜饲粮代谢能水平提供依据。本试验严格按照完全随机区组试验设计进行。选取13周龄北京油鸡600只，公母各占1/2，根据体重一致原则随机分为6组，其中3个组均为公鸡，另外3个组均为母鸡，每组5个重复，每个重复20只。3个组分别为低代谢能组（饲粮代谢能水平为12.20 MJ/kg）、中代谢能组（饲粮代谢能水平为12.64 MJ/kg）、高代谢能组（饲粮代谢能水平为13.08 MJ/kg）。预试期1周，正试期4周（14～17周龄）。结果显示：1）高代谢能组公鸡和母鸡的平均日增重显著高于其他2组（P <0.05），且高代谢能组母鸡的料重比显著低于其他2组（P <0.05），饲粮代谢能水平为13.08 MJ/kg时公鸡和母鸡有较好的生长性能。2）公鸡方面，低代谢能组的腿肌率显著高于其他2组（P<0.05）；母鸡方面，低代谢能组的全净膛率显著高于高代谢能组（P<0.05），翅膀率显著低于高代谢能组（P <0.05）。低代谢能组公鸡和母鸡的皮脂...