无菌人工养殖海水中副溶血弧菌噬菌体与宿主的消长动态

在3组一定浓度的副溶血弧菌菌液中分别加入3种浓度的副溶血弧菌噬菌体,第12天各实验组再次分别加入一定浓度的副溶血弧菌或副溶血弧菌噬菌体,比较各组水体中副溶血弧菌噬菌体及其宿主的动态关系。结果表明,副溶血弧菌噬菌体可使宿主下降至一定浓度并与之保持动态平衡关系,实验组1、2、3中副溶血弧菌浓度从2.43×106 mL-1分别下降至6.2×104、9.2×104、7.46×105 mL-1,并以此浓度范围维持下去。高浓度副溶血弧菌噬菌体与其宿主之间平衡关系的稳定性较好,再次加入副溶血弧菌,高、中浓度组分别在1 d和4 d后副溶血弧菌下降至平衡时的数量级浓度,并持续下去。低浓度组副溶血弧菌组有所上升,并持续下去。再次加入副溶血弧菌噬菌体对副溶血弧菌浓度影响较小(P>0.05)。     

口蹄疫病毒VP1植物表达载体的构建

P1 T重组质粒上含有口蹄疫病毒 (FMDV)GD10分离株的p1cDNA片段 ,以此为模板 ,用PCR方法扩增其中的VP1基因 ,获得大小约 6 40bp的片段。该片段用BglⅡ和BstEⅡ酶切消化后克隆至表达载体 pCAMBIA130 5 .2 ,转化EcoliTOP10感受态细胞。重组质粒经PCR、酶切及序列分析 ,证实VP1基因处于CaMV35S启动子控制 ,且读码框正确     

Shrimp arginine kinase being a binding protein of WSSV envelope protein VP31

Viral entry into the host is the earliest stage of infection in the viral life cycle in which attachment proteins play a key role. VP31 (WSV340/WSSV396), an envelope protein of white spot syndrome virus (WSSV), contains an Arg-Gly-Asp (RGD) peptide domain known as a cellular attachment site. At present, the process of VP31 interacting with shrimp host cells has not been explored. Therefore, the VP31 gene was cloned into pET30a (+), expressed in Escherichia coli strain BL21 and purified with immobilized metal ion affinity chromatography. Four gill cellular proteins of shrimp (Fenneropenaeus chinensis) were pulled down by an affinity column coupled with recombinant VP31 (rVP31), and the amino acid sequences were identified with MALDI-TOF/TOF mass spectrometry. Hemocyanin, beta-actin, arginine kinase (AK), and an unknown protein were suggested as the putative VP31 receptor proteins. SDS-PAGE showed that AK is the predominant binding protein of VP31. An i n vitro binding activity experiment indicated that recombinant AK’s (rAK) binding activity with rVP31 is comparable to that with the same amount of WSSV. These results suggested that AK, as a member of the phosphagen kinase family, plays a role in WSSV infection. This is the first evidence showing that AK is a binding protein of VP31. Further studies on this topic will elucidate WSSV infection mechanism in the future.     

Studies on hemolysis of hemolysin produced by Synechocystis sp. PCC 6803

Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemolysin had not been re-ported.To characterize the action mode of hemolysin produced by the wild-type strain of Synechocystis sp.PCC6803,hemolysis of erythrocytes originating from human,mouse,sheep,rabbit and goldfish was studied.The erythrocytes of mouse,sheep and rabbit were sensitive,while those of human and fish were resistant,to this hemolysin.Using rabbit erythrocytes,it was shown that hemoly-sis occurred in two steps:a binding step within the first 10 min of treatment and a lytic step after 30 min.Both binding and lysis were highly temperature-dependent.Effects of erythrocyte density on hemolysis suggest that the hemolysin might target erythrocytes via a multiple-hit mechanism.In the osmotic protection experiment,all tested osmotic protectants,with molecular diameters ranging from 0.9 ?5.66 nm,failed to effectively inhibit hemolysis.Scanning electron micrographs showed that the hemolysin caused protuberances or echinocytes in rabbit erythrocytes,and then disrupted and ruptured the erythrocytes.Characteristics of hemolysis showed distinct differences from other pore-forming mechanisms,suggesting that this hemolysin might act through a detergent-like or lipase mecha-nism,rather than a pore-forming mechanism.     

草鱼呼肠孤病毒096vp7基因生物信息学分析及其酵母表达载体的构建

草鱼呼肠孤病毒（Grass carp reovirus, GCRV）是草鱼出血病的病原.从患病草鱼体内分离到一株草鱼呼肠孤病毒GCRV 096,经反转录PCR,克隆得GCRV 096 长度为855 bp 的vp7 基因,并分析该基因编码蛋白的特性.结果表明,同一基因型分离株VP7 蛋白间的差异不大,但不同基因型分离株VP7 蛋白间存在很大的差异.对GCRV 096 VP7 蛋白生物信息学分析表明,信号肽位点位于20 氨基酸处,且VP7 含有4个潜在的抗原决定簇.构建GCRV 096 vp7 基因的酵母表达载体pGBKT7-S10,并成功转化至酵母中.     

青海玛多MS7.4地震安西站VP宽频带倾斜仪数据特征分析

青海玛多M_S7.4地震前甘肃省安西县地震监测站VP宽频带倾斜仪出现明显异常变化且记录到该地震的同震波形。结合CTS-1地震计进行融合分析可知,2套仪器同震波形的震相到时基本同步,表明2套仪器具有可靠性和一致性,但震相记录特征存在差异。2021-04-17 VP宽频带倾斜仪数据打破原有倾斜趋势,年变向西北方向倾斜,05-13恢复为向西南方向倾斜,震前记录到倾角急剧增大的方位与震中至台站的方位一致,倾斜旋转变化方向与区域构造应力方向一致。在同时段时均值数据时频谱中,异常时段存在（1.085~4.340）×10^-6 Hz(周期为64~128 h)的低频信号。此次地震中2套仪器的优势频率和时频变化特征存在显著差异,这种差异可能与2套仪器不同的机械结构、频段宽度、采样频率有关。     

临汾台峒体摆数字倾斜仪在忻州五寨4.2级地震前的异常特征分析

2011年3月7日忻州五寨4.2级地震前,位于震中西南方向326.8 km外的临汾台的数字化峒体观测仪器,其记录曲线图出现了不同程度的突变、阶变、抖动现象.其中,SSQ-21石英摆、VPl,VP2宽带倾斜仪的短临异常现象较为明显.通过分析发现:SSQ-2I石英摆、VPl宽带摆、VP2宽带倾斜仪对近场地震有很好的映震性能...     

Studies on Antiviral and Immuno-Regulation Activity of Low Molecular Weight Fucoidan from Laminaria japonica

The antiviral activity in vitro and in vivo and the effect of the immune system of two fucoidan fractions with low molecular weight and different sulfate content from Laminaria japonica(LMW fucoidans) were investigated in order to examine the possible mechanism. In vitro, I-type influenza virus, adenovirus and Parainfluenza virus I were used to infect Hep-2, Hela and MDCK cells, respectively. And 50% tissue culture infective dose was calculated to detect the antiviral activity of two LMW fucoidans. The results indicated that compared with the control group, 2 kinds of LMW fucoidans had remarkable antiviral activity in vitro in middle and high doses, while at low doses, the antiviral activity of 2 kinds of LMW fucoidans was not statistically different from that in the blank control group. And there was no statistically difference between two LMW fucoidans in antiviral activity. In vivo, LMW fucoidans could prolong the survival time of virus-infected mice, and could improve the lung index of virus-infected mice significantly, which have statistical differences with the control group significantly(p 0.01). However, the survival time of the two LMW fucoidans was not statistically significant(p 0.05). In this study, it was shown that both of two LMW fucoidans(LF1, LF2) could increase the thymus index, spleen index, phagocytic index, phagocytosis coefficient and half hemolysin value in middle and high doses, which suggested that LMW fucoidans could play an antiviral role by improving the quality of immune organs, improving immune cell phagocytosis and humoral immunity.     

An improved method for detection of Edwardsiella tarda by loop-mediated isothermal amplification by targeting the EsrB gene

Edwardsiella tarda is a major pathogen in aquatic environments that can cause heavy economic losses. An improved method for quick and accurate detection of E. tarda by loop-mediated isothermal amplification (LAMP) with two additional loop primers was developed by targeting the EsrB gene (EsrB-LAMP). In this method, the Mg 2+ concentration, reaction temperature, and reaction time were optimized to 8 mmol/L, 61°C, and 40 min, respectively. The detection limit with the EsrB gene was as low as 10 copies, which is 100 times more sensitive than that of conventional polymerase chain reaction (PCR). The EsrB-LAMP assay was shown more sensitive and rapid than previously reported LAMP assays targeting the hemolysin gene (hemolysin-LAMP) for detection of E. tarda. The EsrB-LAMP was also highly specific to E. tarda and had no cross-reaction with 13 other strains of bacteria. The assay can be carried out in a simple heating device and the EsrB-LAMP products can be visually detected by adding fluorescent dye to the reaction mixture. Taken together, the improved EsrB-LAMP diagnostic protocol has the potential for detection of E. tarda from indoor and outdoor samples.     

溶藻弧菌疫苗对凡纳滨对虾免疫功能的影响

应用溶藻弧菌疫苗饲喂凡纳滨对虾,测定了弧菌疫苗对凡纳滨对虾免疫指标的影响以及 疫苗的免疫保护作用。结果发现,投喂弧菌疫苗后,溶菌活力在12～20d内迅速增强,在20d时 活力达到最高,VF1组高于VF2和VF3组,但VF2和VF3之间差异不显著;凝集效价在投喂后的 第8天开始升高,在14d时达到最高,实验组的凝集活力均高于对照组,但实验组之间无明显的差 异;而超氧化物歧化酶(SOD)活力和溶血活力变化不明显。以每克饲料含疫苗细胞1.0×109、 1.0×108和1.0×107个的浓度投喂,对凡纳滨对虾的免疫保护率分别为66.7%、46.7%和13.3%。 可见弧菌疫苗对凡纳滨对虾的免疫保护率随疫苗浓度的增大而升高。     

河北VP型宽频带垂直摆倾斜仪映震效能及时频特征分析

利用河北地区8套VP型宽频带垂直摆倾斜仪的连续观测资料,从映震特征及映震能力等方面讨论其映震效能,并对其时频响应特征进行分析。结果显示：1）河北地区宽频带垂直摆倾斜仪的映震能力与震级和震中距正相关,与震源深度负相关（崇礼台除外）;对于同一地震而言,各台站震中距的远近与仪器映震能力相关性不大。2）8套仪器均能记录到较为完整的远震地震波信息,从波形上能够清晰看到P波与S波的起始时刻。3）时频分析结果显示,仪器能够记录到较为完整的面波信号,但受采样率的限制,体波（P波和S波）信号记录不全,无法记录到0.5 Hz以上的信号。4）由于厂家修改了崇礼台仪器的高端截止频率参数,调整后仪器记录的高频成分增多,信噪比降低,对浅源地震的映震效能产生了与其他仪器相反的结论,对该仪器参数的修改结果需作进一步验证。     

VP垂直摆观测背景的功率谱密度特征分析

对正常时段及干扰时段VP垂直摆秒采样数据进行功率谱密度分析,有助于进一步认识观测数据的正常背景和干扰特征,为识别地震异常信息、揭示地壳变化过程提供更加客观的背景参考。     

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VP型垂直摆倾斜仪观测频带的分析及改进

从仪器原理、传递函数和观测数据等方面分析目前在网进行1 Hz采样的VP型垂直摆倾斜仪在2~60 s周期内的频率特性,认为该仪器可以观测到频带内的信号变化,但因处于过渡带,观测幅度较真实幅度削弱约20~40 dB。对VP型垂直摆倾斜仪观测频带进行拓展,使2~60 s周期内的观测信号由之前的过渡带进入到仪器的通频带,结果与相同采样率的其他定点形变仪器的观测效果相同,将地倾斜观测量转换为加速度表达后,其幅频特性符合全球地震背景噪声模型。改进后的VP型垂直摆倾斜仪对于远距离震级不大的同震体波震相记录效果略优于改进前,但受地脉动影响,在识别非地脉动信号时需要采用更精细的方法和手段来提高识别效果。     

A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda

Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry.Therefore,a rapid,reproducible,and sensitive method for detection and quantification of this pathogen is needed urgently.To achieve this purpose,we developed a TaqMan-based real-time PCR assay for detection and quantification of E.tarda.The assay targets the hemolysin activator HlyB domain protein of E.tarda.Our optimized TaqMan assay is capable of detecting as little as 40 fg of genomic DNA per reaction.A standard curve was generated from the threshold cycle values(y) against log10(E.tarda genomic DNA concentration) as x.The intra-and inter-assay coefficient of variation(CV) values were less than 2.06% and 1.05% respectively,indicating that the assay had good reproducibility.This method is highly specific to E.tarda strains,as it shows no cross-reactivity to Edwardsiella ictaluri,a member of the same genus,or to nine other fish-pathogenic bacteria species belonging to three other genera.This sensitive and specific real-time PCR assay provides a valuable tool for diagnostic quantitation of E.tarda in clinical samples.     

Elevation,slope aspect and integrated nutrient management effects on crop productivity and soil quality in North-west Himalayas,India

On farm bio-resource recycling has been given greater emphasis with the introduction of conservation agriculture specifically withclimate change scenarios in the mid-hills of the north-west Himalaya region（NWHR）. Under this changing scenario, elevation, slope aspect and integrated nutrient management（INM） may affect significantly soil quality and crop productivity. A study was conducted during 2009-2010 to 2010-2011 at the Ashti watershed of NWHR in a rainfed condition to examine the influence of elevation, slope aspect and integrated nutrient management（INM） on soil resource and crop productivity. Two years of farm demonstration trials indicated that crop productivity and soil quality is significantly affected by elevation, slope aspect and INM. Results showed that wheat equivalent yield（WEY） of improved technology increased crop productivity by -20%-37% compared to the conventional system. Intercropping of maize with cowpea and soybean enhanced yield by another 8%-17%. North aspect and higher elevation increased crop productivity by 15%-25% compared to south aspect and low elevation（except paddy）. Intercropping of maize with cowpea and soybean enhanced yield by another 8%-15%. Irrespective of slope, elevation and cropping system, the WEY increased by -30% in this region due to INMtechnology. The influence of elevation, slope aspect and INM significantly affected soil resources（SQI） and soil carbon change（SCC）. SCC is significantly correlated with SQI for conventional（R2 = 0.65＊）, INM technology（R2 = 0.81＊） and for both technologies（R2 = 0.73＊）. It is recommended that at higher elevation.（except for paddy soils） with a north facing slope, INM is recommended for higher crop productivity; conservation of soil resources is recommended for the mid hills of NWHR; and single values of SCC are appropriate as a SQI for this region.     

京津唐地区HFMD时空变异分析与影响因子探测

手足口病（HFMD）是一种多发于儿童的常见传染病。近年来,中国HFMD发病人数逐年上升,疾病疫情也日益受到社会广泛关注。虽已有不少相关研究,但对于探测其时空异质性及量化潜在影响因子解释力的研究仍然较少。本文采用地理探测器及贝叶斯时空层次模型,对2009-2013年京津唐地区HFMD发病率的时空异质性进行分析,并量化各影响因子及其两两交互作用对HFMD发病率的解释力。结果表明：① HFMD的相对风险存在时间异质性,其发病风险在春夏季（5-7月）达到峰值,而冬季（12-次年2月）发病风险最低;② HFMD的相对风险存在空间异质性,在经济发达的地区HFMD发病率较高;③ 影响HFMD发病率时间异质性的主要气象因子为平均温度、累积降水、相对湿度,解释力分别为0.38,0.27,0.13,且交互作用都大于独自影响的作用,如平均温度和相对湿度、平均温度和降水,平均温度和风速两两交互的解释力分别为0.43, 0.40, 0.42。通过研究京津唐地区HFMD发病率的时空异质性以及影响因子的量化状况,为本地区HFMD的预防和控制提供理论依据。     

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Effects of drought on net primary productivity: Roles of temperature,drought intensity,and duration

Northeast China has experienced frequent droughts over the past fifteen years. However, the effects of droughts on net primary productivity (NPP) in Northeast China remain unclear. In this paper, the droughts that occurred in Northeast China between 1999 and 2013 were identified using the Standardized Precipitation Evapotranspiration Index (SPEI). The NPP standardized anomaly index (NPP-SAI) was used to evaluate NPP anomalies. The years of 1999, 2000, 2001, and 2007 were further investigated in order to explore the influence of droughts on NPP at different time scales (3, 6, and 12 months). Based on the NPP-SAI of normal areas, we found droughts overall decreased NPP by 112.06 Tg C between 1999 and 2013. Lower temperatures at the beginning of the growing season could cause declines in NPP by shortening the length of the growing season. Mild drought or short-term drought with higher temperatures might increase NPP, and weak intensity droughts intensified the lag effects of droughts on NPP.     

GEOCHEMICAL STUDIES OF Fe,Mn,Co,As,Cr,Sb,Zn,Sc AND V IN SURFACE SEDIMENTS FROM JIAOZHOU BAY

The contents of nearly forty-elements in surface sediments in Jiaozhou Bay were determined using a Neutron Activation Analysis Technique(Grancini,et al.,1976;Li Peiquan et al.,1985,1986;Li Xiuxia etal.,1986).This paper's detailed discussion on only nine elements(Fe,Mn,Co,Cr,Sc,As,Sb,Zn andV)includes their distributions,concentrations,correlationships,material sources,background,etc.Based on Zavaristski's classification method,Fe,Mn,Co Cr and V belong to the second group;As and Sb to theeighth groups:Sc and Zn to the third and sixth groups.It was found that their notably good correla-tionship is mainly due to the similarity of their ionic structures and that their variation is controlled by theFe content(except Mn).The source of sediments is mainly terristrial material,and the composition ofsediment is similar to that of shale and shale+clay.The contents for a large number of ele-ments are within the scope of the background level,but there still is pollution of Zn and Cr,at least in a few stations.