规模化蛋鸡场病原菌溯源与生物安全防控研究

细菌病严重威胁蛋鸡健康,以抗生素为主导的细菌病传统防控方式会导致病原菌耐药性严重、鸡蛋药物残留,影响蛋品安全。要保证蛋鸡产蛋期不用抗菌药物而又能生产出"无菌、无抗"的鸡蛋,是国内外技术难题。本团队以规模化蛋鸡场全封闭鸡舍为研究对象,基于PFGE、MLST、全基因组测序等技术,探明了蛋鸡病原菌的种类及基因组特征;对病原菌的耐药性研究表明,蛋鸡病原菌多重耐药严重,依靠药物防控的方式亟待转变;对规模化蛋鸡场细菌溯源研究结果表明,鸡舍生物媒介(鼠、苍蝇等)和非生物媒介(空气、饮水、饲料、鸡粪等)是蛋鸡病原菌的重要传染来源和传播途径;对规模化蛋鸡场生物安全量化评价结果表明,外部和内部生物安全权重高达92.2%。根据溯源和生物安全权重分析结果,提出了在规模化全封闭蛋鸡舍将蛋鸡细菌病从药物防控转变为生物安全防控的新策略;创新了规模化蛋鸡场细菌病防控的系统技术,实现蛋鸡规模养殖整个产蛋期不用抗生素,为生产"无菌、无抗"安全鸡蛋提供技术支撑。     

Influences of sand cover on erosion processes of loess slopes based on rainfall simulation experiments

Aeolian-fluvial interplay erosion regions are subject to intense soil erosion and are of particular concern in loess areas of northwestern China.Understanding the composition,distribution,and transport processes of eroded sediments in these regions is of considerable scientific significance for controlling soil erosion.In this study,based on laboratory rainfall simulation experiments,we analyzed rainfall-induced erosion processes on sand-covered loess slopes(SS)with different sand cover patterns(including length and thickness)and uncovered loess slopes(LS)to investigate the influences of sand cover on erosion processes of loess slopes in case regions of aeolian-fluvial erosion.The grain-size curves of eroded sediments were fitted using the Weibull function.Compositions of eroded sediments under different sand cover patterns and rainfall intensities were analyzed to explore sediment transport modes of SS.The influences of sand cover amount and pattern on erosion processes of loess slopes were also discussed.The results show that sand cover on loess slopes influences the proportion of loess erosion and that the compositions of eroded sediments vary between SS and LS.Sand cover on loess slopes transforms silt erosion into sand erosion by reducing splash erosion and changing the rainfall-induced erosion processes.The percentage of eroded sand from SS in the early stage of runoff and sediment generation is always higher than that in the late stage.Sand cover on loess slopes aggravates loess erosion,not only by adding sand as additional eroded sediments but also by increasing the amount of eroded loess,compared with the loess slopes without sand cover.The influence of sand cover pattern on runoff yield and the amount of eroded sediments is larger than that of sand cover amount.Furthermore,given the same sand cover pattern,a thicker sand cover could increase sand erosion while a thinner sand cover could aggravate loess erosion.This difference explains the existence of intense erosion on slopes that are thinly covered with sand in regions where aeolian erosion and fluvial erosion interact.     

硒对母猪氧化应激及其后代抗氧化能力的营养调控研究

硒作为的重要微量元素之一,可以通过胎盘和乳组织进行转移。仔猪可以从乳汁中获取硒源,提高硒含量,进而提高仔猪的初生重。同时,硒也能够调节母猪妊娠期间代谢速度,控制妊娠期自由基产生,进而提高胎盘输送营养功能。文章从母猪氧化应激体系角度来解析硒对母猪妊娠期氧化应激的调控机制、硒对哺乳母猪生产性能的调控作用。     

Effects of the severity and timing of basal leaf removal on the amino acids profiles of Sauvignon Blanc grapes and wines

The effects of the severity and timing of leaf removal(LR) on the amino acids of Sauvignon Blanc grapes and wines were studied during the 2017 growing season. High-performance liquid chromatography(HPLC) was used to analyze the amino acids profiles of grape berries and wines. The basal leaves were removed at three time points(40, 56 and 72 days after flowering, named LR40, LR56 and LR72, respectively) at two severity levels(one at which the first, third, and fifth basal leaves of each shoot were removed(50% level); and another at which the first six basal leaves were removed(100% level)). The results showed that leaf removal had little impact on total soluble solids(°Brix), titratable acidity, pH or berry weight. The LR72-50% treated grapes had higher berry weight, titratable acidity and °Brix than those of the other treatments. The highest concentrations of total amino acids and of total amino acids except proline were detected in LR72-50% treated grapes(2 952.58 and 2 764.36 mg L~(-1), respectively); the lowest were detected in LR72-100% treated grapes(2 172.82 and 2 038.71 mg L~(-1), respectively). LR72-50% treatment significantly promoted the synthesis of aspartic acid, serine, arginine, alanine, aminobutyric acid and proline at both severity levels for grapes, the concentrations of all of these amino acids were increased relative to the control concentrations. The LR72-50%, LR40-100% and LR72-100% treated wines had higher total amino acids concentrations and higher concentrations of some individual amino acids, such as arginine, alanine and serine, than did the control wines. Of all the amino acids studied, glycine, tyrosine, cysteine, methionine and lysine were not significantly influenced by the timing or severity basal defoliation in grapes and wines. The present study reveals the effects of the timing and severity of leaf removal on the amino acids profiles of grapes and wines.     

持续低温胁迫下大花蕙兰不同组织器官耐寒响应

采用自然降温处理,分别测定了大花蕙兰地上部分和地下部分中丙二醛(MDA)含量、膜透性(CMP)、可溶性蛋白质(SP)含量、可溶性糖(SS)含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性和过氧化氢酶(CAT)活性等与耐寒性有关的生理指标,比较地上部分和地下部分耐寒差异性,研究低温胁迫对大花蕙兰组培苗生理特性的影响。结果表明:POD活性、可溶性蛋白质含量和可溶性糖含量在反映大花蕙兰组培苗耐寒性方面作用明显,可作为其耐寒性鉴定指标;地上部分和地下部分在耐寒性提高时表现出不一致性,后者比前者表现出更强的耐寒性能。     

白花蛇舌草超微粉对小鼠免疫功能的影响

The assay was aimed to study the effect of Hedyotis diffusa Willd superfines powder on immune function. Three groups of mice were gavaged with the drugs at doses of 1.75,1.25 and 0.75 g/kg once a day for 7 consecutive days, repectively. The mice of control group were administered with the same volume of saline.At the last time, mice were injected into the muscle with Newcastle disease vaccine,and get peripheral blood on days 7, 14 and 21 after the medicine administration. The enhancement of innate immune responses were evaluated by using CCK-8 method, hemagglutination inhibition test, macrophage phagocytic function test and organ coefficient method. The levels of lymphocytes proliferative capacity, serum antibody, peritoneal macrophage phagocytosis and the indexes of immune organs in mice were extremely significantly enhanced (P＜0.01). Hedyotis diffusa dWilld superfines powder could extremely significantly regulate the immunity function of mice (P＜0.01).     

茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建

CRISPR/Cas9技术是一门新兴的基因组定点编辑技术,具有操作简单、高效的优点,可轻松实现对目标基因的敲除、替换和定点突变等操作。该技术刚诞生,就受到了全球生命科学领域研究者的关注,不到3年的时间就已经成功应用于多种动、植物当中。然而CRISPR/Cas9技术在茶树中的应用面临载体构建问题,本文以茶树咖啡碱合成酶为例,联合采用常规PCR、Overlapping PCR和Golden Gate Cloning技术,构建了包含茶树咖啡碱合成酶双靶点的CRISPR/Cas9基因编辑载体,为CRISPR/Cas9介导的基因组编辑技术在茶树中的应用奠定了坚实基础。     

不同施肥方式对红壤蔬菜田氨氧化细菌和氨氧化古菌群落的影响

通过构建氨单加氧酶基因(amoA)克隆文库,研究在红壤蔬菜田上只施用磷钾化肥(PK)、只施氮磷钾化肥(NPK)、施用腐熟有机肥(DNPK)和施用新鲜有机肥(FNPK)等4种不同施肥处理的土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)群落多样性及与土壤脲酶活性的相关性。结果表明:施加有机肥处理(DNPK和FNPK)的蔬菜田土壤的AOB文库和AOA文库OTU数量和Shannon指数高于只施用无机肥(NPK和PK)处理的蔬菜田土壤;DNPK和FNPK处理的土壤优势AOB菌群为多形亚硝化叶菌(Nitrosolobus multiformis),比例分别为88.5%和68.5%,NPK和PK处理的土壤优势AOB菌群为亚硝化单胞菌属(Nitrosospirasp.),比例分别为54.8%和65.5%;DNPK、FNPK、NPK和PK处理土壤优势AOA菌群均为阿伯丁土壤亚硝化细杆菌侯选种(CandidatusNitrosotalea devanaterra),比例分别为90.9%、84.4%、77.8%和45.2%;施加有机肥处理(DNPK和FNPK)的土壤脲酶活性和氨氧化微生物的多样性指数都高于只施用无机肥处理(NPK和PK);AOA群落多样性指数与土壤脲酶活性呈显著正相关,而AOB群落多样性与土壤脲酶活性相关性不显著。总体来看,有机肥比无机肥处理提高了AOA和AOB群落多样性,且AOA在红壤蔬菜田土壤氨氧化过程中起着更为重要的作用。     

黄土沟壑区开采沉陷对地形因子的影响研究

以黄土沟壑区的采煤沉陷区为研究对象,以地面数字高程模型和开采沉陷预计结果为基础数据,基于ArcGIS平台对煤矿开采沉陷区的地形数据进行分析处理,研究开采沉陷前、后地形因子的变化特征,并对不同地形因子之间的相关性进行了分析.研究结果表明:黄土沟壑区地面高程、坡度、坡向、起伏度等各种地形因子均受开采沉陷影响而发生改变,开采沉陷和地形因子之间存在明确的量化关系,而平均高程、起伏度、曲率、沟壑密度等因子与地面坡度之间存在显著的相关性.研究结果有助于揭示煤矿开采沉陷区的地形地貌演变规律,为保护西部矿区的生态环境提供技术支持.     

用于构建双价禽流感DNA疫苗的双目的基因表达质粒的构建及鉴定

为探索构建禽流感双价或多价DNA疫苗的可行性,本研究以本实验室前期研制的禽流感DNA疫苗p CAGGopti HA5为基础[其中含有禽流感病毒(AIV)A/Goose/Guang Dong/1/96(H5N1)的HA基因],将增强型绿色荧光蛋白(eGFP)报告基因作为第二个目的基因,利用双启动子启动、内部核糖体进入位点(IRES)介导以及通过蛋白Linker编码序列连接融合表达两个目的基因的形式,构建了双启动子重组表达质粒pH5-eGFP、IRES介导的重组表达质粒pHIE以及融合表达重组质粒pH6E。并采用脂质体介导将这3种重组质粒分别转染293T细胞,通过间接免疫荧光试验和western blot方法检测HA和eGFP这两种目的蛋白的瞬时表达情况。结果显示,这3种重组质粒均能够正确表达AIV的HA蛋白和eGFP。其中pH5-eGFP表达出两个独立的目的蛋白,而且互相不影响各自的表达和细胞定位;pH6E则以融合形式表达两种目的蛋白,由于HA蛋白具有膜定位信号,所以融合蛋白主要定位于细胞膜上。本研究结果表明,双启动子表达质粒和双目的蛋白融合表达质粒适合用于构建AIV以及其他相关病原的双价DNA疫苗。