Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

LED补光对水稻秧苗素质及其生理特征和产量的影响

采用发光二极管(light-emitting diode,LED)精确调制光谱能量,研究不同光质对水稻秧苗素质(株高、根长和鲜干重)和生理特征(内源激素、抗氧化酶、可溶性蛋白和丙二醛)的影响,探讨生理特征与秧苗素质和补光移栽后产量的耦合关系。水稻秧苗分别在5种光质下进行补光照射:R (100%红光),B (100%蓝光),RB (80%红光+20%蓝光)RBG_12.5(62.5%红光+25%蓝光+12.5%绿光),RBG₂₅(50%红光+25%蓝光+25%绿光),光量子通量均为1 000μmol·m^-2·s^-1,照射30 d (6 h/d)后采样,并插秧至稻田,以无补光作为对照(CK)。与CK相比,LED补充光源显著(P<0.05)影响水稻秧苗的株高、根长和鲜干重。株高和地上部分干重最大值分别出现在R和RBG_12.5处理,而根长和地下部分干重的最大值均出现在RBG_12.5处理。LED补充光源对内源激素、抗氧化酶、可溶性蛋白和丙二醛的影响程度因光质和植物部位而异。与CK相比,RBG_12.5二显著(P<0.05)提高地上部生长素(IAA)含量和可溶性蛋白含量,B显著(P<0.05)提高地上部超氧化歧化酶(SOD)活性、过氧化物酶(POD)活性和丙二醛(MDA)含量;RBG_12.5和RBG₂₅显著(P<0.05)提高地下部IAA含量,RBC₂₅显著(P<0.05)提高地下部可溶性蛋白含量和MDA含量。补光移栽后水稻产量的变化趋势与光质对水稻秧苗壮苗指数的影响趋势一致:RBG_12.5> RBG25> RB> B> R> CK。逐步回归分析发现,壮苗指数与地上部赤霉素(GA_3)含量和IAA含量及地下部IAA含量和SOD活性显著正相关;补光移栽后产量与秧苗地上部IAA含量及地下部IAA含量和过氧化氢酶(CAT)活性显著正相关。LED补充光源通过调节水稻秧苗生理特征影响其素质,且影响效应延续至水稻成熟期。RBG_12.5光源更利于培育水稻壮苗和水稻增产,适宜用作工厂化育秧的补充光源。     

神经介素B及其受体NMBR参与抗A型流感病毒H1N1亚型感染的信号通路

NMB/NMBR通过调节A型流感病毒（IAV/H1N1/PR8）感染诱导的细胞因子表达而参与抗IAV的先天性免疫反应。为探究其发挥抗IAV/H1N1感染的信号通路，本文用PR8和WSN毒株分别感染MLE-12细胞和小鼠，用NF-κB抑制剂BAY11-7028单独或联合NMB处理MLE-12细胞，小鼠后腿肌内注射NMB和NMBRA，采用RT-PCR和qRT-PCR分析NMB、NMBR、IL-6、IFN-α和NP基因表达变化，采用Western blot分析NMB、NMBR、P65/p-P65、IκBα和NP蛋白表达的变化。结果显示，BAY11-7028可促使PR8和WSN感染的MLE-12细胞中NMB、NMBR、IL-6和IFN-α基因表达水平均下降和NP基因表达水平上升，并降低NMB、NMBR和p-P65蛋白表达水平和提升IκBα和NP蛋白表达水平。然而，NMB联合BAY 11-7028诱导PR8或WSN感染后的细胞中IL-6和NP表达出现极显著下降和IFN-α显著上升。此外，NMB抑制PR8和WSN感染的小鼠肺组织内p-P65和NP蛋白表达水平和促进IκBα蛋白表达水平；NMBRA联合NMB抵消NMB对PR8或WSN感染后的这些蛋白表达水平的调节作用。综上表明，NMB/NMBR通过调节PR8和WSN感染的MLE-12细胞和小鼠体内的NF-κB信号通路上P65蛋白磷酸化和IκBα的表达，进而影响下游细胞因子IL-6和IFN-α基因的表达，从而发挥抗IAV/H1N1感染的先天性免疫应答反应。     

外源NO及反向调控PEG胁迫下苜蓿萌发种子抗氧化酶及其同工酶动态的研究

以紫花苜蓿为材料,用一氧化氮供体硝普钠(SNP)及一氧化氮清除剂c-PTIO对苜蓿种子进行浸种处理,采用分光光度法和同工酶电泳技术来研究外源NO及反向调控对PEG胁迫下紫花苜蓿抗氧化酶活性及其同工酶的影响,并探讨NO调控苜蓿种子耐旱性的生理机制。结果表明:PEG胁迫下外施0.1 mmol·L^-1 SNP,第6天时较PEG处理POD活性降低了32.72%;第4天时SOD、CAT活性较PEG处理升高了10.48%、23.60%,有效缓解了PEG对紫花苜蓿萌发中种子的氧化损伤,提高其抗氧化能力。PEG胁迫下添加 c-PTIO抑制了苜蓿萌发期的抗氧化系统活性。从同工酶的谱带数量和强弱来看,POD同工酶各区带活力均随PEG胁迫时间的延长而增加,在第2天酶带只有1条,而第4天酶带呈现9条;SOD和CAT同工酶表达量变化不显著,但酶带强弱有一定变化,S3酶带随处理时间的延长表达量逐渐减弱;CAT同工酶谱带则一直保持2条带,无明显强弱变化。因此,外源NO在PEG胁迫下紫花苜蓿萌发中抗氧化酶快速响应并在维护氧自由基代谢平衡中起重要保护作用。     

柑橘褐斑病菌对不同抗性柑橘品种防御酶活性的影响

以柑橘褐斑病高抗品种Lw8,中抗品种L2,高感品种Lw14为实验材料,通过孢子喷雾法使柑橘褐斑病菌感染柑橘叶片,研究接菌0d、1d、2d、3d、6d、8d、10d时,柑橘叶片中超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、几丁质酶(CHT)酶和β-1,3葡聚糖酶(GLU)含量及活性变化规律与品种抗病性的关系。结果表明,接种前,两个抗病品种Lw8、L2中的POD、SOD、PAL、PPO 、GLU、CHT活性显著高于高感品种Lw14,CAT活性则与高感品种Lw14相近；接种后,3个不同抗性品种中SOD、POD、CAT、PAL、PPO 、GLU、CHT 7种防御酶活性较对照明显提高,但不同品种增加幅度不一致,除CAT外,其余6种防御酶活性均表现为抗病品种Lw8、L2增幅显著高于高感品种Lw14,且两个抗病品种一般在接菌3d内6种防御酶活性迅速升高并达到峰值；而高感品种Lw14防御酶(除CAT外)活性或增幅较小或较两个抗病品种滞后。本研究初步探讨了不同抗性品种接种褐斑病菌后7种防御酶的活性动态变化与柑橘品种抗病性关系,为进一步研究其抗病机理奠定基础。     

黑龙江鱼类开发利用浅析

黑龙江流域地处我国高寒地区,地理环境和鱼类区系组成特殊,形成了许多特有的名贵经济鱼类,为我国的渔业经济发展做出重大贡献。本文简要概述黑龙江流域的鱼类资源、黑龙江省渔业发展现状,以及主要经济鱼类的养殖状况,分析讨论黑龙江鱼类的生物学种质特点,以及目前名优鱼类的市场需求,并针对黑龙江鱼类的开发利用提出了几点建议。     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.     

不同品种藜麦苗期对海拔变化的生理响应

为研究不同品种藜麦幼苗对海拔梯度变化的生理响应机制,本试验以4个藜麦品种—‘陇藜1号’、‘陇藜2号’、‘陇藜3号’及‘陇藜4号’为材料,选取甘南州临潭县3个不同海拔(2380 m,2580 m,2780 m)分布区为试验地。通过测定幼苗生物量及生理生化指标,分析不同藜麦品种幼苗在不同海拔试验区的生理响应机制,为建立藜麦品种和环境相适宜的评价体系提供参考依据。结果表明,随海拔升高,藜麦叶片叶绿素、可溶性蛋白以及抗坏血酸过氧化物酶(APX)含量逐渐降低,海拔2780 m最低;超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)逐渐升高,在海拔2780 m达到最大值;超氧阴离子(O2·-)产生速率呈现先升高后降低的趋势。不同藜麦品种幼苗生物量随着海拔的升高逐渐降低。说明高海拔下太阳辐射强度增大、温度降低,导致藜麦幼苗植株抗氧化酶活性升高以清除多余活性氧(ROS)自由基,使得植株能够适应高海拔环境而正常生长,但生物量较低海拔仍有降低。因此,不同品种藜麦在低海拔地区受到的胁迫较低,虽然在高海拔区域胁迫加剧,但藜麦幼苗可以正常生长并进行干物质的积累。     

Exogenous xylanase improves growth,protein digestibility and digestive enzymes activities in Nile tilapia,Oreochromis niloticus,fed different ratios of fish meal to sunflower meal

A 3 × 2 factorial experiment was designed to evaluate the effect of different ratios of fish meal (FM): sunflower meal (SFM) with or without exogenous xylanase supplementation on growth, feed utilization, digestive enzymes activities, apparent digestibility, intestinal and liver morphology and chemical composition of Nile tilapia, Oreochromis niloticus. Three isonitrogenous (329.80 g/kg of crude protein) and isoenergetic (18.46 MJ/kg gross energy) experimental diets were formulated as SFM₁ (FM:SFM = 2:1), SFM₂ (FM:SFM = 1:1) and SFM₃ (FM:SFM = 1:2) based on protein content. Each diet was supplemented with 0 or 0.5 g/kg of exogenous xylanase and was fed to triplicate groups of twelve fish (with initial weight, 1.31 ± 0.02 g) for 84 days. After 84 days of feeding period, the highest weight gain, specific growth rate, protein efficiency, protein productive value and the best feed conversion ratio were recorded in fish fed either SFM₁ or SFM₂ supplemented with exogenous xylanase. Whereas lowest growth performance was recorded in fish fed SFM₂ and SFM₃ un‐supplemented with xylanase. The highest activities of chymotrypsin, trypsin, lipase, amylase, alkaline phosphatase and cholecystokinin were observed in fish fed SFM₁ and SFM₂ diets supplemented with xylanase. The highest ADCs of dry matter, protein, lipid and digestible energy were recorded in fish fed SFM₁ and SFM₂ diets supplemented with exogenous xylanase. Supplementation of exogenous xylanase improved muscularis mucosa thickness, height of mucosal folds and enterocytes of intestinal fish. Addition of exogenous xylanase increased the calcium and phosphorus retention. Results of this study indicated that the addition of exogenous xylanase to diet containing high inclusion level of sunflower meal improved growth, digestive enzymes, nutrient digestibility, histological morphometric of liver and intestine and nutrient retention.     

苏云金芽胞杆菌JQD117菌株对韭蛆体内几种酶活性的影响

为明确苏云金芽胞杆菌JQD117对韭蛆幼虫蛋白酶和解毒酶活性的影响，测定比较了感染Bt后幼虫体内胰蛋白酶、胰凝乳蛋白酶、乙酰胆碱酯酶、谷胱甘肽-S-转移酶和羧酸酯酶的活性。首先采用室内生物活性测定方法明确了菌株JQD117对韭蛆3龄幼虫72 h的LC₅₀为2.8070×10⁷cfu/mL，然后采用10×LC₅₀、1×LC₅₀和0.1×LC₅₀三个浓度饲喂感染韭蛆幼虫，定期取样测定韭蛆体内5种酶活性，结果表明以较高浓度（1×LC₅₀和10×LC₅₀）感染韭蛆幼虫后体内蛋白酶和解毒酶活性变化较大，而以低浓度（0.1×LC₅₀）感染韭蛆幼虫后体内蛋白酶活性变化较小。其中，以1×LC₅₀和10×LC₅₀浓度感染韭蛆幼虫后，胰蛋白酶和乙酰胆碱酯酶活性在24~36 h和6~60 h与对照相比均显著升高；类胰凝乳蛋白酶和羧酸酯酶活性在6~60 h与对照相比均受到显著抑制。以0.1×LC₅₀浓度感染韭蛆幼虫后，胰蛋白酶活性只在36 h时与对照相比显著升高；乙酰胆碱酯酶活性在12~48 h时与对照相比显著升高；羧酸酯酶活性只在6 h与对照相比受到显著抑制；类胰凝乳蛋白酶活性与对照相比均无显著变化。谷胱甘肽-S-转移酶活性在三种感染浓度下与对照相比均无显著变化。可见，感染JQD117对韭蛆体内蛋白酶和解毒酶活性均产生了不同程度的影响，且随感染浓度的升高而增强，扰乱了韭蛆正常的生理代谢和对外源毒素的分解，本文为Bt防治韭蛆应用和开发提供了理论指导。