复合侵染水茄的两种菜豆金色花叶病毒属病毒基因组结构特征分析

为明确水茄Solanum torvum植株叶片邹缩、褪绿是否由菜豆金色花叶病毒属病毒侵染引起,从云南省西双版纳傣族自治州田间采集具有疑似感染症状的水茄植株叶片样品,应用菜豆金色花叶病毒属病毒简并引物和特异性引物进行PCR扩增、克隆和测序,通过生物信息软件分析比较其核苷酸序列特征,并对其进行系统发育分析。结果显示,从采集的疑似病叶中共克隆获得了5条菜豆金色花叶病毒属病毒DNA-A全序列和3条DNA-B全序列,经全序列分析发现,侵染水茄的2种菜豆金色花叶病毒属病毒分离物分别属于中国南瓜曲叶病毒(squash leaf curl China virus,SLCCNV)和野茼蒿黄脉病毒(Crassocephalum yellow vein virus,CraYVV)。SLCCNV水茄分离物的基因组具有典型的菜豆金色花叶病毒属病毒双组分结构特征,与来自泰国的SLCCNV分离物(AB330078)亲缘关系最近,相似性最高达到99.0%;CraYVV水茄分离物的基因组具有典型的菜豆金色花叶病毒属病毒单组分结构特征,与来自云南省景洪市的CraYVV分离物(EF165536)亲缘关系最近,相似性最高达到97.6%。表明水茄是这2种菜豆金色花叶病毒属病毒的新寄主,并首次发现双组分和单组分菜豆金色花叶病毒属病毒可复合侵染水茄。     

Effect of Fermented Chinese Medicine Preparation by Bacillus subtilis on Growth Performance,Biochemical Indices,Antioxidant Parameters and Ability of Anti-infection of Common Carp

The objective of this study was to evaluate of the growth performance, serum biochemical indices, liver antioxidant parameters and ability of anti-infection with Aeromonas hydrophila by adding the fermented Chinese medicine by Bacillus Subtilis to the carp fish feed, and to find out the appropriate supplemental level of fermented Chinese medicine in diets. Carp fingerling with an initial body weight of (35±0.5) g were set up to 6 experimental diets supplemented with 0 (control group), 0.1‰, 0.5‰, 1‰, 2‰ and 3‰ fermented Chinese medicine for 56 days, respectively. Each group had 30 fish and set up 3 repetitions. After the feeding trail,the fish were challenged with Aeromonas hydrophila to calculate the relative percentage survival (RPS).The results showed as follows:Supplementing fermented traditional Chinese medicine by Bacillus subtilis, the weight gain rate (WGR), specific growth rate (SGR), condition factor (CF), the activities of lysozyme (LSZ), complement 3 (C3),total protein (TP),total anti-oxidation capacity (T-AOC),total superoxide dismutase (T-SOD) and catalase (CAT) in serum of common carp were enhanced at different degrees, while the feed coefficient (FCR)and malondialdehyde (MDA) in serum were declined at different degrees.When the fermented Chinese medicine supplemental levels were 2‰ and 3‰,the effects were significant (P<0.05)(except for C3, ALP and Cr).The experimental results of the virulence of common carp infected with pathogen Aeromonas hydrophila showed that,the fermented herbal could obviously improve the ability of anti-infection, the immune protective rate of the group with 2‰, 3‰ doses of the fermented Chinese medicine could reach 77.23% and 74.63%, respectively. From what had been discussed above, the group with 2‰ and 3‰ doses of the fermented Chinese medicine had significant effect on the growth performance, serum biochemical indices, liver antioxidant parameters and Aeromonas hydrophila anti-infection ability. Considering the cost of adding fermented Chinese medicine, the recommendations for the best additive quantity was 2‰.     

A faster inoculation assay for Armillaria using herbaceous plants

Armillaria (honey fungus) is a virulent necrotrophic pathogen that causes Armillaria root disease. Conventional Armillaria inoculation assays use young saplings as hosts and consequently are cumbersome, frequently conducted outdoors, and take many years from establishment to analysis of infection. We have developed and evaluated a faster inoculation assay for Armillaria that uses herbaceous plants as hosts, is carried out in controlled conditions, and reduces experimental durations to 3 months. Plant species of known susceptibility to Armillaria and comparisons between virulent A. mellea and opportunistic A. gallica were used to validate the assay. Mortality and diagnostic symptoms of Armillaria root disease such as epiphytic rhizomorphs and mycelial fans were used to assess levels of infection. We also attempted to reduce assay preparation time by substituting woody inocula with agar inocula, but typical symptoms of Armillaria root disease were only observed on plants infected with woody inocula. Through our assay, we identified five new potential herbaceous hosts of Armillaria: Kniphofia hirsuta, Hordeum vulgare, Lobelia cardinalis, Nicotiana tabacum and Helenium hoopesii – further expanding the extensive list of plants with susceptibility to Armillaria and suggesting infection of herbaceous species may be more widespread than currently acknowledged.     

Stilbene oligomer phytoalexins in grape as a response to Aspergillus carbonarius infection

Stilbenes are grapevine phytoalexins elicited by biotic and abiotic agents; Aspergillus carbonarius is a widespread ochratoxin A producing fungus present in warm conditions, such as in Southern Italy. To increase the knowledge on biosynthesis of stilbene oligomers induced by A. carbonarius infection, grape berries of the Southern Italian grape cv. Negro Amaro were inoculated. Significant increase of trans-resveratrol and resveratrol dimers and oligomers, such as caraphenol, E-ε-viniferin, ω-viniferin, δ-viniferin, α-viniferin, E-miyabenol C, and two tetramers, was observed, and concomitant decrease of glycoside derivatives. These findings improve the knowledge on the phytoalexin production as response against this pathogen.     

PRV、PCV-2、PRRSV与HPs混合感染的核酸检测

为确定青海省某规模化发病猪场的发病原因,在临床诊断的基础上对采集的临床组织病料进行了13种常见病原体的核酸检测,并对检测结果进行了序列鉴定.结果表明,该发病猪场存在猪伪狂犬病毒、猪圆环病毒2型、猪繁殖与呼吸综合征病毒与副猪嗜血杆菌的混合感染.研究结果提示,利用分子生物学技术对发病猪场的病原进行快速鉴定,是病源快速诊断和防控的有效途径.     

亚成体东北虎感染巴氏杆菌死亡一例

2009年1月11日,北方森林动物园发生一起自繁自养的亚成体东北虎感染巴氏杆菌死亡病例。本病例从发病到死亡共5 d时间,病程较短,以食欲下降至废绝,饮水量不变,精神沉郁,口腔腺体分泌增多、流涎,呈滴水样,腹部疼痛感明显为特征,口服诺氟沙星和多酶片治疗无效后改用肌注头孢哌酮钠和庆大霉素治疗,3 d后出现神经症状抽搐死亡。剖检肝脏表面有大量的针尖大小黄白色点状坏死灶。根据临床症状、病理剖检和实验室检查,确诊为巴氏杆菌感染。     

1株感染多种致病性大肠杆菌噬菌体的生物学特性研究

旨在测定此前从断奶前仔猪粪样中分离得到的1株大肠杆菌噬菌体C6在致病性大肠杆菌上的生物学特性,并比较该噬菌体在不同致病菌上的感染特性。利用电镜形态观察和基因组测序确定其分类,用点滴法和双层平板法测定其在致病性大肠杆菌上的宿主谱,通过噬菌斑形态、最佳感染复数（MOI）、成斑率（EOP）、吸附率、一步生长曲线以及抑菌曲线等来评价噬菌体在多株致病菌上的感染特性。结果显示：噬菌体C6可感染1株非致病性大肠杆菌和4株致病性大肠杆菌,包括大肠杆菌O157（E.coli W1）、2株产志贺毒素大肠杆菌（STEC W3、STEC W5）和1株肠致病性大肠杆菌（EPEC 143）。通过比较噬菌体C6在4株致病菌上的各项感染特性指标发现：噬菌斑大小和成斑率E.coli W1>EPEC 143>STEC W3>STEC W5;吸附率E.coli W1>EPEC 143、STEC W3、STEC W5;最佳MOI EPEC 143、STEC W3、STEC W5>E.coli W1;液体LB中生长能力和抑菌能力E.coli W1>STEC W3>EPEC 143、STEC W5。噬菌体C6为T4样噬菌体,可感染多株致病性大肠杆菌,在不同致病菌上的感染特性存在差异,但均能显著抑制致病菌的生长。     

干扰素刺激基因对羊口疮病毒在OFTu细胞中增殖的抑制效应

羊口疮病毒（ORFV）是重要的人兽共患病病原,不仅严重危害养羊业,而且威胁人类健康。干扰素刺激基因（stimulator of interferon genes,STING）作为细胞的DNA感受器,在机体天然免疫中起重要作用。为探索STING在ORFV感染中的作用及其对病毒复制的影响,本研究构建了ORFV感染羊胚胎鼻甲细胞（OFTu）的模型,分析了ORFV感染细胞后对STING及其相关基因的动态表达,探索了STING基因在干扰表达和过表达状态下对ORFV在细胞上增殖的影响。结果表明,ORFV感染OFTu细胞后,STING、cGAS、TBK1、IRF3、IRF7、IL-6、IFN-β、IL-1β和TNF-α的转录明显升高。OFTu细胞过表达STING可导致RIG-1、DDX41、IFI16、IRF3、IRF7、IL-6、TNF-α、IFN-α和IFN-β等基因转录上调。OFTu细胞在STING过表达状态下感染ORFV可介导TBK-1、IRF3、IFN-β和TNF-α的转录升高,抑制ORFV的复制;在STING表达干扰的状态下,ORFV感染OFTu细胞降低了TBK-1、IRF3、IFN-β和TNF-α的转录,增加了ORFV的复制。这表明STING蛋白能够增强抗病毒细胞因子的表达,抑制ORFV在OFTu细胞中的增殖,研究结果为深入理解STING在羊口疮病毒感染和复制中的作用提供了科学的理论依据,也为深入探索ORFV感染和致病的分子机制提供了基础数据。     

汝南县奶牛附红细胞体病的流行病学调查与综合防控

目的 附红细胞体病是由附红细胞体（Eperythrozoon）寄生于人及动物的红细胞表面、血浆及骨髓中引起的一种人畜共患传染病。方法 为调查汝南县奶牛附红细胞体病流行情况,2020年3月到2021年2月选取748 头奶牛采用现场调查、询问和实验室诊断等方法进行流行病学调查。结果 汝南县奶牛附红细胞体病感染率为10.03%,其中,泌乳牛阳性率最高,为13.02%,青年牛次之,为10.20%,犊牛最低,为5.91%,7、8月为发病高峰期,发病率分别为14.55%、14.28%,1月发病率最低,为6.67%。结论 调查为汝南县奶牛附红细胞体病防治提供了科学参考。