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.     

Caecal intussusceptions and typhlocolitis in horses with severe Gastrodiscus aegyptiacus infestation

The intestinal trematode Gastrodiscus aegyptiacus (G. aegyptiacus) has been recognised in equids around the world for many years, but its pathogenicity is yet to be confirmed. This report describes seven cases of severe G. aegyptiacus infestation, including six cases of caecal intussusception.     

紫花苜蓿9个盐响应相关基因表达模式

紫花苜蓿(Medicago sativa)是世界上种植面积最大、应用最广泛的豆科牧草。由于其耐盐性中等,其在我国北方地区的产量和种植受到土壤盐渍化的限制。因此提高紫花苜蓿的耐盐性具有重要的科学和生产意义。为此,以龙牧801紫花苜蓿(M.sativa‘Longmu 801’)为试验材料,采用实时荧光定量PCR技术分析了不同浓度NaCl胁迫下9个盐胁迫蛋白质组筛选出的盐响应相关基因的表达模式。结果显示,处理时间和处理浓度对9个基因的相对表达量均有显著性影响,表明这9个基因均在紫花苜蓿盐胁迫应答中发挥着一定作用。处理1h时,G6PI、ABP19a、Trx-h1、PR bet 1、FBPA、6PGDH和ALDH这7个基因的相对表达量在不同NaCl胁迫浓度处理的紫花苜蓿中均显著上调,RRM和GDPD在NaCl处理2h后开始上调。除G6PI基因,其他8个基因在0.4%NaCl处理下相对表达量显著高于0.2%和0.8%NaCl处理。这些基因参与糖代谢、信号转导和胁迫响应。以上结果表明,紫花苜蓿的耐盐性极其复杂,涉及到多基因的表达和代谢通路的调控。研究结果有助于全面研究并了解紫花苜蓿的盐响应相关基因的表达模式,促进紫花苜蓿耐盐分子育种。     

Impact of cold-season grazing and supplementary feeding on rumen metabolic parameters and micro-organisms in the ewes of Gansu Alpine Fine Wool Sheep北大核心CSCD

An experiment was conducted to scientifically identify the appropriate feed for Gansu Alpine Fine Wool ewes in order to change the animals' breeding mode, which currently can only be mated at the age of 2.5 years due to insufficient nutritional supply. The experiment selected 48 12-month-old ewes and randomly divided them into group a (Diet 1), group b (Diet 2), group c (Diet 3), and a control group with no supplementary feeding. The feed test was run for 90 days. The rumen pH of groups a and b were very significantly higher (P<0.01) or group c significantly lower (P<0.05) than control group. The rumen ammonia nitrogen, protein nitrogen and total nitrogen of groups a, b and c were all higher than control group (P<0.01). The total volatile fatty acids of group a and b were all higher than the control group (P<0.01), group b significantly higher than group c (P<0.05). The molar ratio of acetic acid and propionic acid in group a was very significantly higher than control group (P<0.01), group b and c were significantly higher than control group (P<0.05). The molar ratio of butyric acid in group a was very significantly higher than group c (P<0.01). The ratio of acetic acid and propionic acid in group b was significantly lower than control group (P<0.05). The number of several rumen fiber-degrading bacteria and protozoa in groups a, b and c were very significantly (P<0.01) or significantly (P<0.05) higher than the control. In conclusion, this study shows that supplementary feeding can positively affect the rumen metabolic parameters and the number of several rumen microorganisms in Gansu Alpine Fine Wool ewes. In general, the effects of Diet 1 were the strongest, while Diet 2 also produced notable improvements. © 2018, Editorial Office of Acta Prataculturae Sinica. All rights reserved.     

草鱼呼肠孤病毒生长特性研究

以草鱼呼肠孤病毒(GCRV)873毒株为试验材料,建立了GCRV荧光定量PCR检测方法;用GCRV873株感染CIK细胞及草鱼,通过对病毒RNA复制水平、子代病毒含量以及细胞病变等情况进行监测,研究其在CIK细胞及草鱼体内的生长特性。在CIK细胞中,GCRV感染后12 h即可检测到子代病毒,36 h时,所有细胞被感染;72 h时,病毒滴度达到最高,为10~(6.75) TCID_(50)/mL。在草鱼的肝、脾、肾中,均能检测到GCRV RNA,其中肾脏中含量最高,其次是肝脏,脾脏中最少。本试验为研究GCRV致病机理及制备细胞灭活疫苗奠定了试验基础。     

一起未经执业兽医注册从事诊疗活动案引发的思考

2017年成都市办理的一起未经兽医执业注册而进行犬只声带切除的行政处罚案件,引发了笔者对动物诊疗机构监督执法中"动物诊疗"范畴界定、动物诊疗许可证申请对象探究、"一事不再罚"原则的适用、"责令停业诊疗活动"与"责令停产停业"区别等问题的思考。本文以法学理论和相关法律规定为基础,对成都市动物诊疗机构执法实践的疑点与难点进行分析,提出执法人员应更准确适用法律法规,并呼吁尽快建立全国执业兽医查询系统的建议。     

信号肽对溶血素在大肠杆菌中跨膜转运的影响

信号肽在引导外源蛋白的跨膜转运过程中具有重要作用。本试验通过PCR方法获得带有信号肽序列和不带有信号肽序列的溶血素基因,构建表达载体在大肠杆菌中进行诱导表达。经过溶血试验和免疫印迹试验证明缺失了信号肽的重组溶血素不能被跨膜转运至细胞外;带有信号肽的重组溶血素能够跨膜转运大肠杆菌细胞外。本试验为猪链球菌2型基因缺失弱毒疫苗的研究、细菌信号肽以及基因工程重组蛋白的可溶性表达进行了有益探索。     

如何做好重大动物疫病防控消毒药品的政府采购

本文指出了政府在消毒药品采购中存在高素质专业人才匮乏,采购方评标标准不一,低价中标忽视质量,不同品牌消毒剂功效差异大,国家标准有待完善等问题,建议政府采购以质量为先,尽快出台消毒药品采购指南,加强采购人员专业素质培养等。此外,政府采购的消毒药品应选择高效速效、安全低毒、无三致、使用成本低、适用大环境使用的药品,这有助于重大动物疫病防控工作的开展。     

Copper‐Induced Oxidative Stress and Responses of the Antioxidant System in Roots of Medicago sativa

The effects of various copper (Cu) concentrations on the antioxidative system in the roots of Medicago sativa were explored. The results indicated that the Cu content of the roots reached a value of 854 μg g^?1 DW at 10 μm Cu and a value of 4415 μg g^?1 DW at 100 μm Cu, suggesting that M. sativa has better ability to tolerate and accumulate Cu than other Cu‐bioaccumulators, and is a potential plant for phytoremediation. Treatment with Cu resulted in a significant increment in the levels of H₂O₂, O₂˙^? and OH˙. The reduced form of ascorbate and glutathione reached a peak at 30 μm Cu, and was followed by a sharp depletion to a lower level than that of the control. In contrast, the levels of the oxidised forms of ascorbate and glutathione showed a progressive increment with increasing Cu concentrations, suggesting that the antioxidant system was unable to cope with Cu stress at higher Cu levels. Under the Cu concentrations tested, the activity of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) increased at lower Cu concentrations, and then decreased, reaching a maximum at 30 μm of Cu for APX and GR, at 10 μm for CAT, whereas the activities of guaiacol peroxidase (POD, EC 1.11.1.7) were gradually increased with increasing Cu concentrations. PAGE analysis of superoxide dismutase (SOD, EC 1.1.5.1.1) revealed that one band is a Mn‐SOD and five bands are identified as Cu, Zn‐SOD, whereas Fe‐SOD isoforms were not found in the roots of alfalfa. Cu at 10–100 μm increased the intensity of constitutive isozymes of CAT, APX and POD, whereas it decreased the intensity of isozymes of glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49) significantly. The activities of lipoxygenases (LOX, EC 1.13.11.12) were gradually augmented with increasing Cu concentrations, demonstrating that LOXs are probably involved in production of lipid hydroperoxides and superoxide anion. There was a continuous and pronounced enhancement in the activity of esterase (EST, EC 3.1.1.1) in roots treated with 10–30 Cu μm , whereas EST activity in roots exposed to above 30 μm Cu declined, suggesting that EST plays a protective role under lower Cu concentrations stress.     

南方地区高水分玉米安全度夏技术探讨

根据水分的高低,对新收购的高水分玉米分别储存在露天囤和高大平房仓中,将露天囤里的玉米通风降水后转入高大平房仓储存,再对玉米通风降温,并采取仓房密封、悬挂防晒网、空调控温等配套措施控制粮温,期间采取磷化铝多次补药熏蒸杀灭粮堆内害虫及霉菌,使玉米安全度夏。