Plasma exosomes have been identified to be involved in embryo development and implantation. In this study, the plasma exosomes of seven pregnant cows on day 16 and seven non-pregnant cows were collected. Exosome miRNA profiles were revealed by RNA sequencing technology and the abundance of exosome miRNA between them were compared. Seven miRNAs were significantly differently expressed (p-value < 0.05, |log2(FoldChange)| > 1), with five miRNAs upregulated and two downregulated in the pregnant cow group. The most significant miRNA was bta-miR-136 (downregulated in pregnant cows, p-value < 0.005). A total of 5828 target genes were computationally predicted for the seven miRNAs. GO and KEGG enrichment analyses showed these targets were significantly involved in several biological processes and pathways associated with embryo implantation and endometrial development, such as cell adhesion, cell junction, focal adhesion, Ras signaling pathway and Rap1 signaling pathway. This study identifies seven differentially expressed plasma exosomes miRNAs on day 16 of pregnancy in cows, with bta-miR-136, bta-miR-199b, and bta-miR-382-3p emerging as candidate miRNAs for future studies on maternal-fetal communication and embryo implantation.

In March 2022, a large-scale dairy farm in Henan Province suffered from neonatal calf diarrhea. On the basis of epidemiological investigation, observation of clinical symptoms and pathological dissection, the fresh feces of 6 diarrhea calves were examined for parasite eggs, Cryptosporidium parvum, bovine rotavirus, coronavirus, and K99 antibody against Escherichia coli. The blood samples were examined for bovine viral diarrhea virus antigen, and the tissue samples were isolated for bacteria. The results showed that 6 bovine rotavirus antigens were positive, and the positive rate was 100%. Coronavirus antigen was positive in 4 cases and negative in 2 cases（66.7%）. Other pathogens were negative. According to the clinical symptoms, pathological changes and laboratory test results, these cases were confirmed as calf diarrhea caused by mixed infection of bovine coronavirus and rotavirus. According to the diagnosis results, comprehensive prevention and control measures such as improving feeding management, fluid infusion, convergence and diarrhea prevention were taken, which effectively controlled the epidemic situation and provided a reference for clinical prevention and treatment of calf diarrhea.

The aim of this study was to detect the novel copy number variation (CNV) locus of NCAM2 gene in Chinese Holstein, and to analyze the effect of the novel CNV locus in NCAM2 gene on milk composition traits. The novel CNV locus of NCAM2 gene in 310 Chinese Holstein was detected by real-time quantitative fluorescent PCR (qPCR) and association analysis was performed between the novel CNV locus in NCAM2 gene and milk composition traits in Chinese Holstein. There are three CNV types of NCAM2 gene in Chinese Holstein: gain (increased copy number), median (normal copy number) and loss (deleted copy number). Statistical analysis revealed that there was a significant association between CNV types and milk fat rate (p < 0.05). Moreover, we also discovered that the milk production and milk protein rate of gain type is higher than that of loss type, but that of mediate type is lower than that of loss type. However, in terms of somatic cell score, loss type is higher than that of gain type, but that of mediate type is lower than that of gain type. These observations suggested that gain type can be used as a candidate molecular genetic marker of milk fat rate.

旨在分析不同因素和SMG9基因拷贝数变异对奶牛繁殖性能的影响,以期为提升奶牛繁殖性能和探究SMG9基因拷贝数功能提供参考.收集河南某牛场200头已孕奶牛的配种次数、产犊后首次配种天数和空怀天数等繁殖性状数据,统计父亲、年龄、胎次、配种员和配种季节等因素,并通过荧光定量PCR法鉴定每头牛SMG9基因的拷贝数类型.通过一般线性模型将SMG9基因的拷贝数类型与繁殖性状进行关联分析,并分析不同因素对繁殖性能的影响.结果:父亲、年龄、胎次、配种季节和SMG9拷贝数变异会显著影响该奶牛群体的繁殖性状(P＜0.05).其中:1胎牛配种次数显著高于2胎牛,秋季配种次数显著高于春季和冬季(P＜0.05),冬季产犊后首次配种天数高于秋季(P=0.064),1胎的空怀天数显著高于2胎(P＜0.05).SMG9基因的拷贝数变异与配种次数和空怀天数显著相关(P＜0.05),多拷贝型个体繁殖性能优于正常型和缺失型.研究结果可为牛繁殖性能提升提供参考,并提示SMG9基因的拷贝数可作为奶牛繁殖性能选育的分子标记.

Intrauterine exosomes have been identified to be involved in the embryo development and implantation. The aim of this study was to explore the role of miRNAs in intrauterine exosomes in bovine pregnancy. Intrauterine exosomes were collected from uterine flushing fluids of three donor and three recipient Xianan cows 7 days after fertilization. Intrauterine exosomes miRNAs were extracted and the exosomal miRNAs expression levels were analyzed. Sixty miRNAs differed significantly in their amounts between donors and recipients (p-value < 0.05, |log2(FoldChange)| > 1). Twenty-two miRNAs were upregulated and 38 downregulated in the group of donor cows. The bta-miR-184 was the most significant (PBenjamini-Hochberg < 0.001). A total of 9,775 target genes were predicted using the 60 miRNAs. GO and KEGG analysis showed that the target genes were enriched in several biological processes or pathways associated with embryo implantation and endometrial development, such as cell adhesion, cell junction, focal adhesion, and Rap1 signaling pathway. Our findings suggest that, in cattle early pregnancy stage, these differently expressed miRNAs in intrauterine exosomes involved in embryo implantation and endometrial development, which may exert a significant effect and influence the uterine microenvironment for embryo implantation. These results could provide reference for screening and exploring the intrauterine exosomal miRNA affecting embryo implantation.

[目的]为了分析河南三种肉牛超数排卵效果.[方法]分别选择夏南牛、郏县红牛、南阳牛作为试验母牛,采用两次PG+促卵泡素(FSH)的方法对试验母牛进行超数排卵,观测试验母牛获得总胚胎数、可用胚数分析超数排卵效果.[结果]结果显示,夏南牛可用胚9.86枚/头,南阳牛可用胚7.71枚/头,郏县红牛为6.75枚/头,可用胚中夏南牛最高与郏县红牛和南阳牛有显著性差异.可用胚率郏县红牛最高79.41％,显著高于夏南牛和南阳牛.[结论]结果表明,PG+FSH法可以用于夏南牛、郏县红牛和南阳牛的超数排卵.

随着我国经济水平迅速增长,肉牛产业生产需求也越来越大.截至2015年底,我国自主培育了4个优秀肉牛品种,为肉牛产业带来了一定的经济效益.但是,由于我国肉牛产业起步较晚,新品种虽然相比于原来的肉牛品种有进步,但仍然面临许多问题.本文就夏南牛、延黄牛、辽育白牛、云岭牛4个自主培育品种的生产现状、存在问题和对策进行论述,并对肉牛产业发展前景进行了展望.

Copy number variations (CNVs) were similar to single nucleotide polymorphisms (SNPs) and insertion-deletion (InDel), regarded as genetic variations in many species. CNV is defined as the variable change of DNA segment length compared with the reference genome, including gains or losses from 50 bp to several mega bases. The functions ofUSP16gene are diverse, such as regulating the cell cycle, DNA damage, histone H2A deubiquitination or mitotic nuclear division. To analyze the relationship between CNV ofUSP16gene and milk traits in Chinese Holstein, we used qPCR to detect the individuals of Chinese Holstein (n = 180). The results showed that the effect ofUSP16gene CNV on daily milk yield and fat percentage had significant difference (p < 0.05). The gain was the advantage type in daily milk yield and the loss was the advantage type in fat percentage. Therefore, CNV ofUSP16gene is an important factor of milk traits in Chinese Holstein. Meanwhile, it may be used as a molecular marker for assisted selection of milk traits in Chinese Holstein, which provides a theoretical basis for the genetic improvement of cow breeds in China.

为了给双肌肉牛品种的培育提供新的遗传素材,拟利用CRISPR-Cas9基因编辑技术制备牛MSTN基因编辑胚胎。从采集到的86对牛卵巢中收集卵母细胞进行成熟培养以及体外受精,并使用在线软件设计能够在体外高效编辑牛MSTN基因的gRNA序列,将验证后所得的体外切割活性最高的gRNA与Cas9 mRNA的混合物显微注射牛的受精卵,培养至囊胚后,利用T7EI核酸内切酶法检测囊胚中MSTN基因的编辑情况,然后将基因编辑阳性的囊胚样品进行测序验证。结果表明,牛胚胎体外培养平均囊胚率为21.28%;在设计的多条gRNA序列中,gRNA5的体外切割活性最高,为96.00%;将Cas9 mRNA和gRNA5的混合物显微注射受精卵并培养至囊胚后,T7EI核酸内切酶检测表明囊胚MSTN基因切割阳性率为15.40%;对MSTN基因编辑囊胚阳性样品测序表明,在gRNA5靶位点存在部分片段缺失。综上,说明利用CRISPR-Cas9基因编辑技术成功获得了牛MSTN基因编辑胚胎。

本研究的目的是分析郏县红牛生长性状遗传基础,筛选与生长性状相关的候选基因.运用SLAF—seq简化测序技术,通过全基因组关联分析,筛选出3个与郏县红牛生长性状相关的SNP位点,rs449748996位点与18月龄体高显著相关(P=9.88E— 13),rs210024569与6月龄体高潜在显著相关(P=9.53E-08),rs42177491与12月龄胸围潜在显著相关(P=2.71E-07).在每个SNP周围1Mb区域内筛选相关基因,并对之进行GO数据库分析,发现rs449748996位点所在的LOC100337124基因可以作为18月龄体高性状的候选基因;rs210024569位点所在的C6orf106基因可以作为6月龄体高性状的候选基因;rs42177491位点所在的JAM3基因能否作为12月龄胸围的候选基因还需要进一步验证.

This study was conducted to study the effects of Chinese herbal medicine additives on dry matter intake,milkperformance and blood biochemical index of dairy cows under heat stress. According to the similar principles in milk yield,lactation day,birth order,body condition score,60 healthy Holstein cows were selected. 60 Holstein cows were randomlydivided into 4 groups with 15 per group. The trial diets were supplementation of 100,200 and 300 g/head · d Chinese herbalmedicine additives into the basal diets. The experiment was conducted for 9 weeks,including a 1 week adaptation period and a8 week treatment period. The results showed as follows :Compared with the groups of 0 and 100 g/head · d,dry matter intakeand milk yield of 200 and 300 g/head · d were significantly increased(P < 0.05). Chinese herbal additives have no significanteffect on milk composition of dairy cows under heat stress(P > 0.05).thyroid hormone and cortisol concentration of testgroup were significantly increased(P < 0.05),and the highest concentration was in the 200 g/head · d group. Superoxidedismutase,glutathione peroxidase activities of test group were significantly increased(P < 0.05),Malondialdehyde contentwere significantly decreased(P < 0.05). In summary,the best effect was obtained with the Chinese herbal medicine additivesapplication in the concentration of 200 g/head · d,it can significantly improve dry matter intake and milk performance.

近年来,动物福利在国际上得到了广泛关注,但是我国关于动物福利的认识依旧比较落后.畜禽产品安全问题以及国际动物福利壁垒引发学者们从动物福利的角度开始思考.我国畜禽业生产经过了三十年的迅猛发展,现阶段已基本满足了人们对畜禽产品数量上的需求,随着经济水平发展和人们生活水平的提高,畜禽产品的主要矛盾已经发生了改变,由原先的数量需求转变为数量与质量并存,从"吃得饱"转变为"吃的健康""吃的营养".另一方面,国际上关于动物福利贸易壁垒问题的突出,严重制约着我国畜禽产品的出口.因此,发展动物福利成为我国亟待解决的问题,良好的动物福利也是保障畜禽产品质量安全的重要途径之一.

中国是中药生产及使用大国,随之产生的中药渣废弃物数量巨大.传统方式处理中药渣不但容易产生环境污染,还造成资源严重浪费.因此,将中药渣应用到畜禽生产中,将其变成畜禽饲料资源,为中药渣的合理利用提供了新的途径.本文主要综述中药渣的主要营养成分、营养价值及其在主要畜禽动物猪、牛、羊和鸡的应用情况和作用效果,旨在为中药渣饲料资源的开发利用提供一定的参考.

奶牛子宫内膜炎是发生在牛子宫内膜上的炎症,为了探索奶牛子宫内膜炎发生时血液生化指标的变化特点,本实验以患子宫内膜炎奶牛为研究对象,与正常奶牛做对比实验.结果显示:患牛Ca、P显著下降,其他指标没有显著变化;2例炎症严重的病牛,谷草转氨酶AST和磷酸激酶CK远远高于平均值,呈离群状态.这些指标变化为子宫内膜炎的诊断治疗提供参考.

This study was aimed to constract growth differentiation factor 9 (GDF9) lentiviral vector,which was stably expressed in goat primary fibroblast.The CDS of sheep GDF9 gene was cloned by gene synthesis,the CDS was 1 362 bp,encoding 453 amino acids.After double enzyme digestion and ligation,the GDF9 fragment was sub-cloned into an empty lentiviral vector.The GDF9 lentiviral vector and the packaging plasmids were co-transfected into 293T cells,and the lentivirus with titer of 1 × 106 TU/mL was obtained.The goat primary fibroblasts were transfected with GDF9 lentivirus,the red fluorescence could be observed in more than 60 ％ of the cells,suggesting the prepared lentivirus had high infection efficiency to goat primary fibroblasts.After purom ycin screening,all cells were able to observe red fluorescence.Real-time quantitative PCR analysis showed that the GDF9 expression level was higher than that of control group.The results indicated that the goat primary fibroblasts stably expressing GDF9 were successfully obtained.This results might lay a foundation for the function study of GDF9 and the goat germplasm resources innovation in the future.

分娩对母牛是一个巨大的应激过程,在这个过程中可能会带来一系列问题,如何作好母牛的产后管理与疾病防控,促进母牛产后子宫恢复,实现一年一胎,对提高牛场繁殖水平意义重大.文章就母牛产后管理关键技术及避免母牛产后出现繁殖障碍的问题进行论述.

为了探讨季节变化对夏南牛血液生理生化指标的影响，本试验选用20头24月龄左右未孕健康母牛，于春季（4月）、夏季（7月份）、秋季（10月份）、冬季（1月份）四个季节分别采集颈静脉血测定13项生理生化指标值。结果表明：白细胞数夏季值与冬季值之间差异显著（ P＜0．05）；碱性磷酸酶及葡萄糖夏季值与秋季值之间差异显著（P＜0．05），夏季值与春季值和冬季值之间差异极显著（P＜0．01）；甘油三酯夏季值与春季值之间差异显著（P＜0．05），其余9项差异不显著。同时对结果进行了分析，以期为夏南牛对季节温度的适应性提供基础数据。

为了解河南省及周边地区规模化猪场猪传染性胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)对磺胺类药物的耐药情况,本研究从河南省及周边地区49个规模化猪场的肺脏和气管中分离出79株疑似APP菌株,血清型鉴定结果显示,分离菌株为1、2、3、5、7、8、9、10型APP,采用纸片扩散法对分离株进行了耐药表型鉴定,同时采用PCR方法对磺胺类药物耐药基因进行了检测.药敏试验结果显示,APP对磺胺类药物的耐药率为100％ (79/79).根据GenBank中登录的序列,设计了3对特异性引物,对磺胺类药物的耐药基因Sul1、Sul2、Sul3进行扩增检测.耐药基因检测结果显示,分离的79株细菌中,Sul2基因检出率为100％ (79/79),Sul1基因的检出率为79.75％(63/79),Sul3基因的检出率为89.87％ (71/79),Sull和Sul2基因同时检出率为70.89％(56/79),Sul1和Sul3同时检出率为62.03％ (49/79),Sul2和Sul3基因同时检出率为77.22％(61/79).

In order to understand the serotype and drug resistance of porcine contagious Actinobacillus pleuropneumoniae(APP),apair of primers was designed according to GenBank database to amplify specific 950 bp fragment,and the molecular identification and antimicrobial susceptibility test of APP serotype 3 were investigated.The results showed that the PCR product sequences were more than 99% homology with the APP serotype 3published in GenBank.The isolated strains were highly resistant and multiple drug resistance.The molecular identification and antimicrobial susceptibility test of APP serotype 3provided the basis for the identification,diagnosis and prevention of porcine contagious pleuropneumonia.

近几年我国肉牛业集约化、规模化程度有了很大提高,但相对于鸡、猪、奶牛业来说,肉牛业的发展还是起步晚、发展慢,特别是近几年由于多方面原因,肉牛养殖业经济效益比较差,甚至亏损,严重打击了肉牛养殖者的积极性,致使国内肉牛存栏量和出栏量锐减.分析原因,除了社会性因素外,肉牛繁殖率低下也是制约肉牛业发展的重要因素.