血清中IBV特异性IgG及总IgG含量变化规律的研究

本研究采用间接酶联免疫吸附试验对人工感染鸡传染性支气管炎病毒的SPF鸡血清中特异性抗体及总IgG含量进行检测并比较其相关性.结果表明感染IBV-M41第1 d、3 d、5 d、7 d、10d、14d、21 d、28 d的血清中特异性抗体水平随感染天数增加而升高,于第14天达到最高,之后略呈降低趋势,与健康对照比较差异显著(P＜0.01);总IgG含量较健康对照组增幅小,差异显著(P＜0.01).同时,两者的消长趋势基本一致.     

Identification and characterisation of microRNAs and Piwi-interacting RNAs in cockerels’ spermatozoa by Solexa sequencing

1. There has been substantial research focused on the roles of microRNAs (miRNAs) and Piwi-interacting RNAs (piRNAs) derived from mammalian spermatozoa; however, comparatively little is known about the role of spermatozoa-derived miRNAs and piRNAs within breeding cockerels’ spermatozoa.

2. A small RNA library of cockerels’ spermatozoa was constructed using Illumina high-throughput sequencing technology. Unique sequences with lengths of 18–26 nucleotides were mapped to miRBase 21.0 and unique sequences with lengths of 25–37 nucleotides were mapped to a piRNA database. A total of 1311 miRNAs and 2448 potential piRNAs were identified. Based on stem-loop qRT-PCR, 8 miRNAs were validated.

3. Potential target genes of the abundant miRNAs were predicted, and further Kyoto Encyclopedia of Genes and Genomes database (KEGG) and Gene Ontology (GO) analyses were performed, which revealed that some candidate miRNAs were involved in the spermatogenesis process, spermatozoa epigenetic programming and further embryonic development.

5. GO and KEGG analyses based on mapping genes of expressed piRNAs were performed, which revealed that spermatozoal piRNAs could play important regulatory roles in embryonic development of offspring.

6. The search for endogenous spermatozoa miRNAs and piRNAs will contribute to a preliminary database for functional and molecular mechanistic studies in embryonic development and spermatozoa epigenetic programming.     

Multimarker and rare variants genomewide association studies for bone weight in Simmental cattle

Bone weight, defined as the total weight of the bones in all the forequarter and hindquarter joints, can reflect somebody conformation traits and skeletal diseases. To gain a better understanding of the genetic determinants of bone weight, we used a composite strategy including multimarker and rare‐marker association to perform genomewide association studies (GWAS) for that character in Simmental cattle. Our strategy consisted of three models: (i) A traditional linear mixed model (LMM) was applied (Q+K‐LMM); (ii) single nucleotide polymorphisms (SNPs) with p‐values less than .05 from the LMM were selected to undergo the least absolute shrinkage and selector operator (Lasso) in the second stage (LMM‐Lasso); (iii) genes containing two or more rare SNPs were examined by performing the sequence kernel association test (gene‐based SKAT). A total of 1,225 cattle were genotyped with an Illumina BovineHD BeadChip containing 770,000 SNPs. After the quality‐control procedures, 1,217 individuals with 608,696 common SNPs and 105,787 rare SNPs (with 0.001 < minor allele frequency [MAF] <0.05) remained in the sample for analysis. A traditional LMM successfully mapped three genes associated with bone weight, while LMM‐Lasso identified nine genes, which included all genes found by traditional LMM. Only a single gene, EPHB3, surpassed the significance threshold after Bonferroni correction in gene‐based SKAT. In conclusion, based on functional annotation and results from previous endeavours, we believe that LCORL, RIMS2, LAP3, PRKAR2B, CHSY1, MAP2K6 and EPHB3 are candidate genes for bone weight. In general, such a comprehensive strategy for GWAS may be useful for researchers seeking to probe the full genetic architecture underlying economic traits in livestock.     

Comparative expression profile of microRNAs and piRNAs in three ruminant species testes using next‐generation sequencing

microRNA (miRNA) and piwi‐interacting RNA (piRNA) are two classes small non‐coding regulatory RNAs that play crucial roles in multiple biological processes such as spermatogenesis. However, there are no published studies on conjoint analysis of miRNA and piRNA profiles among cattle, yak and their interspecies (the dzo) using sequencing technology. Next‐generation sequencing technology was used to profile miRNAs and piRNAs among those three ruminants to elucidate their functions. A total of 119, 14 and six differentially expressed miRNAs were obtained in cattle vs. dzo, cattle vs. yak and yak vs. dzo comparison groups, while there were 873, 1,065 and 1,158 differentially expressed piRNAs in those three comparison groups. The expression of three miRNAs was validated in the three ruminants, and the results suggested that the miRNA expression profiles data could represent actual miRNA expression levels. Moreover, the putative targets of differentially expressed miRNAs were predicted by their own genome, it is worth to note that both the cattle and yak genome were used for dzo, then the targets were subjected to GO enrichment and KEGG pathway analysis, revealing the likely roles for these differentially expressed miRNAs in spermatogenesis. In conclusion, this study provided a useful resource for further elucidation of the miRNAs and piRNAs regulatory roles in spermatogenesis. It may also facilitate the development of therapeutic strategies for dzo reproduction research.     

放牧对武功山草甸土壤微生物生物量及酶活性的影响

以武功山山地草甸为研究对象,通过分析不同放牧强度对土壤微生物生物量碳氮以及酶活性的影响,旨在为退化草甸修复提供理论依据。结果表明:1)不同土壤微生物生物量碳氮含量表现为0-20cm土层20-40cm土层,除20-40cm土层轻牧和中牧的土壤微生物生物量氮无显著差异外(P0.05),随放牧强度增加,土壤微生物生物量碳氮含量均显著降低(P0.05);2)可溶性碳氮含量在0-20和20-40cm土层间无显著差异(P0.05),均表现为在0-20cm土层轻牧显著大于中牧和重牧(P0.05);3)碱解氮和易氧化碳含量表现为在不同土层,轻牧和中牧含量均显著高于重牧(P0.05);4)土壤β-葡萄糖甘酶、β-N-乙酰氨基葡萄糖甘酶和脲酶活性表现为在不同土层下,轻度和中牧均显著高于重牧(P0.05);5)相关性分析表明,不同放牧强度土壤β-葡萄糖甘酶、β-N-乙酰氨基葡萄糖甘酶和脲酶活性与微生物生物量碳氮均呈显著性正相关(P0.05)或极显著正相关(P0.01)。     

广东省广州市活禽批发市场从业人员H7N9流感“知信行”现状调查

为了解广东省广州市活禽批发市场从业人员对H7N9流感的认知、态度和行为,也为制定针对性的健康教育措施提供依据,2017年4—5月对广州市全部6个活禽批发市场的435名从业人员进行了问卷调查。调查结果显示:318名密切接触人员和117名非密切接触人员对H7N9流感相关知识的总知晓率分别为80.59%和69.60%;在"发现对禽高致病性的H7N9流感病毒""病死禽应该进行深埋或焚烧等无害化处理""外省养殖场发生过H7N9流感疫情"方面,密切接触者的知晓率显著高于非密切接触者(P0.03);获取H7N9流感知识的途径主要有培训,以及电视、网络和海报等宣传媒介;绝大多数市场从业人员认为政府有能力防控H7N9流感,对政府采取的H7N9流感防控措施及对目前的"1110"制度感到满意,认为活禽集中屠宰可行(P0.01);从业人员在预防H7N9流感时,接触禽类后用肥皂或洗手液洗手的占比最高(83.45%),其次是戴手套(69.20%)和穿水鞋(63.91%),而戴口罩(39.77%)和用消毒液消毒手(35.86%)的占比较低。调查认为,广州市活禽批发市场从业人员的H7N9流感知识知晓率较高,但个人防护措施实施不全面。调查结果提示,应充分利用培训,以及电视和网络等优势媒介,强化微信和微博等新兴媒介,加强对H7N9流感防控知识的宣传。     

湖北省引入第三方检测机构提升动物产地检疫水平实践

根据规定,产地检疫中对怀疑患有检疫规程规定病种及临床检查发现其他异常情况,以及跨省调运要求实验室检测的动物,需经实验室检测合格后,官方兽医才能出具动物检疫合格证明。湖北省开展试点,引入第三方检测机构,为养殖企业提供相关动物疫病的实验室检测服务,借此提升动物产地检疫技术含量,取得了较好效果。本文介绍了湖北省引入第三方检测机构的试点情况,分析了引入第三方开展动物疫病实验室检测的意义并提出了有关思考,以期为各地提供借鉴。引入第三方检测机构,有助于提升动物产地检疫的技术水平,减少动物疫病跨区域传播风险,提升养殖企业的动物疫病防控主体责任意识,弥补动物疫病预防控制机构检测资源与能力的不足。因此,建议正确认识第三方检测机构的作用;适时修订《动物防疫法》及配套规章;借鉴国外经验,探索建立有效的第三方检测机制;将第三方检测机构的检测服务范围拓展到散养户和小型养殖企业,从而整体提升动物产地检疫水平。     

美国国家动物卫生实验室网络组建与发展现状

2002年,为提升动物外来病入侵和生物恐怖袭击的应对能力,美国以联邦、州立和大学动物卫生诊断实验室为基础,组建了国家动物卫生实验室网络(NAHLN)。十多年来,NAHLN几乎覆盖了美国所有公立动物疫病诊断实验室,网络体系趋于完善;通过联邦和各州持续投资,NAHLN实验室强化了生物安全设施、标准化快检技术、质量体系、电子报告系统、人员素质和应急监测预案等6方面建设,能力大幅提升;按照早发现、快反应、有序恢复(无疫)的理念,设置了风险监测、应急监测、无疫监测3种模式,对10余种优先防范和控制病种实施分类监测,在发现疫情和证实无疫方面发挥了重要作用,HANLN的名字由此得到了美国政府、国会和行业的认可。同时,HANLN也面临着经费支持不足、被动监测组织性不够等问题。     

2018年安徽省部分地区A型禽流感流行率横断面研究

为了解安徽省A型禽流感病毒感染情况,2018年4—5月,在安徽省北部、中部和南部7个市,随机选取各类型场点173个,在每个场点内随机采集35份咽喉/泄殖腔和环境拭子样品,使用荧光RT-PCR进行A型、H5亚型、H7亚型流感病毒核酸检测。结果显示：在4个养殖场、3个活禽批发市场和17个农贸市场检出了A型禽流感病毒,场点阳性率分别为2.9%（4/140）、75.0%（3/4）、80.9%（17/21）,未检出H5和H7亚型流感病毒核酸;A型禽流感群体流行率为11.2%（95%CI：6.5%~15.8%）,个体流行率为1.2%（95%CI：0.9%~1.5%）;活禽交易市场内,禽群感染A型禽流感病毒的风险较高（OR=136,95%CI：32.7~549.4）。结果表明,A型禽流感病毒在养殖和交易环节存在一定的污染,而在活禽交易市场的污染更严重,是今后禽流感防控的重点,需继续加强活禽交易市场的管理和消毒灭源工作。     

两种诊断制品对临床样品中布鲁氏菌抗体的检测比较

本研究以国家标准《动物布鲁氏菌病诊断技术》(GB/T 18646-2018)中的试管凝集试验方法为确诊金标准,对353份临床牛、羊和猪血清样品进行确诊,然后分别采用虎红平板凝集试验抗原和抗体检测试纸条,对该353份临床血清进行检测,比较两种诊断制品在布鲁氏菌病初筛中的差异。经统计分析发现,虎红平板凝集试验抗原敏感性为100%(23/23),特异性为97.88%(323/330),与确诊结果间的Kappa值为0.86;布鲁氏菌抗体检测试纸条敏感性为100%(23/23),特异性为99.39%(328/330),与确诊结果间的Kappa值为0.95,两种方法的重复性试验结果均一致。试验结果证明,布鲁氏菌抗体检测试纸条和布鲁氏菌虎红平板凝集试验抗原都能满足布鲁氏菌病初筛的需求;由于布鲁氏菌抗体检测试纸条的储存、运输和操作的便捷性,在部分地区或特殊环境下可替代布鲁氏菌虎红平板凝集试验抗原用于布鲁氏菌病初筛。