Novel reassortant H5N6 highly pathogenic influenza A viruses in Vietnamese quail outbreaks

Avian influenza A H5N6 virus is a highly contagious infectious agent that affects domestic poultry and humans in South Asian countries. Vietnam may be an evolutionary hotspot for influenza viruses and therefore could serve as a source of pandemic strains. In 2015, two novel reassortant H5N6 influenza viruses designated as A/quail/Vietnam/CVVI01/2015 and A/quail/Vietnam/CVVI03/2015 were isolated from dead quails during avian influenza outbreaks in central Vietnam, and the whole genome sequences were analyzed. The genetic analysis indicated that hemagglutinin, neuraminidase, and polymerase basic protein 2 genes of the two H5N6 viruses are most closely related to an H5N2 virus (A/chicken/Zhejiang/727079/2014) and H10N6 virus (A/chicken/Jiangxi/12782/2014) from China and an H6N6 virus (A/duck/Yamagata/061004/2014) from Japan. The HA gene of the isolates belongs to clade 2.3.4.4, which caused human fatalities in China during 2014–2016. The five other internal genes showed high identity to an H5N2 virus (A/chicken/Heilongjiang/S7/2014) from China. A whole-genome phylogenetic analysis revealed that these two outbreak strains are novel H6N6-like PB2 gene reassortants that are most closely related to influenza virus strain A/environment/Guangdong/ZS558/2015, which was detected in a live poultry market in China. This report describes the first detection of novel H5N6 reassortants in poultry during an outbreak as well as genetic characterization of these strains to better understand the antigenic evolution of influenza viruses.     

Secondary sexual traits and semen characteristic of chicken germline chimeras

Birds obtained by embryo engineering are used to study embryo development and to produce transgenic birds. As this method of producing birds still generate strong emotions of the public opinion head ornaments, testes and semen characteristics of sex chimera roosters were examined to check whether they differ from chickens obtained by non‐manipulated methods. Measurements of head ornaments, testes and semen were correlated with each other. Semen quality factor (SQF) was calculated, as well as the level of fluctuating asymmetry (FA) of bilateral traits (wattles and testes). Positive correlation was found for comb width and wattle length and comb thickness and sperm concentration. Semen characteristics and FA did not exceed the level encounter in other chicken lines. Results obtained indicate that germline chimeras are similar in appearance of secondary sexual traits, and semen and testes characteristics to chickens produced in non‐manipulated way.     

Rapid detection of contagious ecthyma by loop-mediated isothermal amplification and epidemiology in Jilin Province China

The aim of this experiment was to develop a loop-mediated isothermal amplification (LAMP) assay and to research the recent epidemiology of contagious ecthyma in Jilin Province, China, using the assay. A LAMP assay targeting a highly conserved region of the F1L gene was developed to detect contagious ecthyma virus (CEV). Three hundred and sixty-five cases from 64 flocks in 9 different areas of Jilin Province, China, from 2011 to 2014 were tested using the LAMP assay. The results showed that the sensitivity of the LAMP assay was 100 copies of the standard plasmid, which is 100-fold higher than the sensitivity of PCR. No cross-reactivity was observed with capripoxvirus, fowlpox virus, foot-and-mouth disease virus serotype O, foot-and-mouth disease virus serotype Asia I and bluetongue virus. The average positive rate was 19.73% (72/365), and the positive rate was highest in lambs aged 1–6 months. Our results demonstrated that CEV infection was very widespread in the flocks of Jilin Province and that the LAMP assay allows for easy, rapid, accurate and sensitive detection of CEV infection.     

常见计算机病毒的防范

分析了计算机常见的四种病毒,根据其固有特点提出了相应的防范方法及建议。     

表达牛病毒性腹泻病毒E2蛋白的重组乳酸菌的构建及蛋白免疫原性分析

试验旨在构建能表达牛病毒性腹泻病毒（Bovine viral diarrhea virus，BVDV）E2抗原蛋白的重组乳酸乳球菌（Lactococcus lactis），为进一步研制BVDV乳酸菌口服活载体疫苗奠定基础。将BVDV E2基因克隆后测序，根据乳酸乳球菌的密码子偏嗜性进行优化，再将优化的基因片段插入表达载体pNZ8148中，并电转化乳酸乳球菌NZ9000感受态细胞，构建重组乳酸菌pNZ8148-E2/NZ9000，经1 ng/mL乳链菌肽诱导表达后，对菌体物进行了SDS-PAGE和Western blotting分析。将重组乳酸菌pNZ8148-E2/NZ9000口服免疫6~12月龄健康犊牛，在免疫后不同时间点采集血液样品并分离血清，用间接ELISA方法检测抗体水平。结果显示，PCR扩增到了1 149 bp的目的片段，乳酸菌密码子偏嗜性优化后，GC含量从45.28%变为34.30%。重组质粒pNZ8148-E2经酶切鉴定插入片段与预期大小相符，在菌体裂解物中出现大小约42 ku的条带，与预期蛋白大小一致，且该蛋白可与BVDV E2抗体反应。在免疫犊牛的血清中检测到特异性抗BVDV E2蛋白的抗体。本研究结果表明，表达BVDV E2蛋白的重组乳酸菌口服免疫可诱导犊牛产生特异性的体液免疫反应，该重组菌具有较好的免疫原性。     

Ethyl acetate fraction of flavonoids from Polygonum hydropiper L. modulates pseudorabies virus-induced inflammation in RAW264.7 cells via the nuclear factor-kappa B and mitogen-activated protein kinase pathways

Pseudorabies virus (PRV) infection leads to severe inflammatory responses and tissue damage, and many natural herbs exhibit protective effects against viral infection by modulating the inflammatory response. An ethyl acetate fraction of flavonoids from Polygonum hydropiper L. (FEA) was prepared through ethanol extraction and ethyl acetate fractional extraction. An inflammatory model was established in RAW264.7 cells with PRV infection to evaluate the anti-inflammatory activity of FEA by measuring cell viability, nitric oxide (NO) production, reactive oxygen species (ROS) release, and mRNA expression of inflammatory factors, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Its functional mechanism was investigated by analyzing the phosphorylation and nuclear translocation of key proteins in the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Our findings indicate that PRV induced inflammatory responses in RAW264.7 cells, and the responses were similar to that in lipopolysaccharide (LPS)-stimulated cells. FEA significantly suppressed NO synthesis and down-regulated both expression and secretion of COX-2, iNOS, and inflammatory cytokines (P<0.05 or P<0.01). FEA also reduced NF-κB p65 translocation into the nucleus and decreased MAPK phosphorylation, indicating that the NF-κB/MAPK signaling pathway may be closely related to the inflammatory response during viral infection. The findings suggested the potential pharmaceutical application of FEA as a natural product that can treat viral infections due to its ability to mitigate inflammatory responses.     

快速灵敏的甘蔗线条花叶病毒免疫学检测体系的建立

 甘蔗花叶病广泛存在于我国甘蔗种植区,严重影响甘蔗产业的高质量发展。近年来甘蔗线条花叶病毒在蔗区肆虐,尽管针对其的血清学检测技术已经建立,但是快速、准确、高通量的检测方法亟待发掘。本研究制备了^SCSMVCP的抗血清,特异性高,与引起甘蔗花叶病的另两种病原 (高粱花叶病毒和甘蔗花叶病毒) 间没有血清学交叉反应。基于该多克隆抗体,建立了直接抗原包被的ELISA、斑点杂交、Western blot和基于多抗的免疫试纸条检测技术。开发的免疫试纸条检测技术能快速、准确、高通量应用于田间病毒鉴定。本文提供了基于血清学的快速、准确、高通量,且便捷的甘蔗线条花叶病毒检测技术,有助于我国蔗区甘蔗花叶病的监测与防控。     

云南小麦品种（系）锈病和赤霉病抗性功能基因的KASP标记检测

利用5个锈病成株期抗性基因的KASP标记Sr2_ger9 3p、Lr34jagger、CSTM4_67G、Lr68-2、VPM_SNP和抗赤霉病基因Fhb1的KASP标记TaHRC-KASP,对云南省育成的42个小麦品种（系）进行检测,旨在筛选出含有目标基因的优异小麦种质,为云南省持久抗病小麦新品种（系）的选育提供材料。结果表明,4个材料含兼抗型成株抗锈病基因Lr34/Yr18/Sr57,频率为9.52%;6个品种（系）含兼抗型成株抗锈病基因Lr67/Yr46/Sr55,频率为14.29%;7个材料含抗慢叶锈病基因Lr68,频率为16.67%;含兼抗型成株抗锈病基因Sr2/Yr30和成株抗叶锈基因Lr37的材料各有1个,频率均为2.38%;未检测出含抗赤霉病基因Fhb1的品种（系）。云麦69、云麦75、云麦56、宜麦1号和宜麦3号等兼有2个成株期抗锈病基因,可作为今后云南持久抗锈病育种的抗源材料。