绵羊BMP15基因B2突变可视化检测技术的建立

骨形态发生蛋白15（bone morphogenetic protein 15,BMP15）基因突变对绵羊排卵率、产仔数以及繁殖力有很大影响,本研究旨在建立快速可视化检测BMP15基因B2突变的技术。将改良的扩增阻滞突变系统（amplification refractory mutation system,ARMS）与核酸染料SYBR Green Ⅰ结合,针对BMP15基因B2突变设计ARMS特异性引物,使引物下游3'端的最后一个碱基为A,并在下游引物3'端的第3位碱基处设计额外的错配,以提高引物的特异性;经ARMS PCR扩增后,向产物中加入SYBR Green Ⅰ,通过肉眼观察PCR管内的颜色变化来检测BMP15基因的B2突变。经测序发现所采集的样本均为野生型,因此,利用重叠延伸PCR构建BMP15基因B2突变型模板,测序结果显示BMP15基因B2突变型模板构建成功。ARMS PCR结果显示,ARMS特异性引物能够只扩增突变型模板,而不会扩增野生型模板,且扩增效果良好。ARMS PCR扩增后加入SYBR Green Ⅰ发现已知野生型模板与阴性对照一致,呈SYBR Green Ⅰ原始颜色橙黄色,而已知突变型模板呈亮绿色,且色差明显,肉眼可辨。用建立的可视化检测BMP15基因B2突变的技术检测50个小尾寒羊基因组DNA （随机向其中20个样本中加入构建的BMP15基因B2突变型模板）,将可视化检测的突变型样本编号与混合样本时记录的编号对比,结果表明该技术能够准确检测绵羊BMP15基因的B2突变,且准确率高达100%。将构建的BMP15基因B2突变型的模板进行梯度稀释,对本试验可视化检测体系的灵敏度进行检测,发现ARMS可视化检测技术的灵敏度达到0.006 ng/μL。因此,本研究建立的技术能够可视化地检测绵羊BMP15基因B2突变,且准确率高,有望为高繁殖力绵羊的选育提供技术支持。     

A pioneering survey of deep-diving and off-shore cetaceans in the northern South China Sea

A pioneering boat-based survey was conducted in 2019, to gather baseline information regarding the presence, composition, relative abundance, and spatial distribution of deep-diving and off-shore cetaceans in the northern South China Sea (SCS). A total of 27 sightings comprising at least 8 cetacean species were recorded during the 13-day survey, including 5 deep-diving species (i.e. Risso's dolphin [Grampus griseus], short-finned pilot whale [Globicephala macrorhynchus], sperm whale [Physeter macrocephalus], Cuvier's beaked whale [Ziphius cavirostris], and an unidentified beaked whale [either the ginkgo-toothed beaked whale, Mesoplodon ginkgodens, or Deraniyagala's beaked whale, Mesoplodon hotaula]), as well as 3 off-shore dolphins (i.e. pantropical spotted dolphin [Stenella attenuate], striped dolphin [Stenella coeruleoalba], and Fraser's dolphin [Lagenodelphis hosei]). With the exception of pantropical spotted dolphins, all other species were sighted and recorded at sea in the northern SCS for the first time. The pantropical spotted dolphin was the most frequently sighted species, comprising 30% of the total sightings. Deep-diving cetaceans were mainly sighted in the northern Xisha Archipelago, whereas off-shore dolphins were distributed across the survey area. The pantropical spotted dolphin was observed in aggregations of more than 100 individuals and nearly all encountered species included calves; these findings suggested that the survey area functions as an important feeding and calving ground for various cetacean species. This pioneering survey provides fundamental information regarding cetacean fauna in the northern SCS and highlights the need to strengthen research and conservation efforts concerning these species.     

Green tea polyphenols supplementation alters immunometabolism and oxidative stress in dairy cows with hyperketonemia

Peripartal cows often experience negative energy balance, and are therefore prone to suffering from metabolic diseases such as hyperketonemia, which causes financial losses in dairy farms. This study aimed to investigate the effect of green tea polyphenol (GTP) supplementation during the periparturient period on production performance, oxidative stress and immunometabolism in dairy cows with hyperketonemia. One hundred Holstein cows were assigned to GTP (0.2 g/kg DM; n = 50) or control (without GTP; n = 50) group based on body weight, previous milk yield, and parity on d 15 before expected parturition. Subsequently, 10 cows with hyperketonemia were selected from each group, according to blood β-hydroxybutyric acid (BHBA) concentration between 1.2 and 2.9 mmol/L from 2 to 3 d postpartum. All cows were fed a close-up diet and a lactation diet with or without GTP supply from 15 d prepartum until 30 d postpartum. Milk and blood samples were obtained from 20 cows selected with hyperketonemia on 10, 20, and 30 d postpartum. Compared with control cows, greater milk yield and lower somatic cell count were observed in GTP cows. The GTP group had lower concentrations of BHBA, free fatty acids, cholesterol, triglyceride, reactive oxygen species, malondialdehyde, and hydrogen peroxide, greater concentrations of glucose, lower activities of aspartate aminotransferase, alanine aminotransferase, and glutamyl transpeptidase, alongside greater activities of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity. Additionally, GTP supplementation up-regulated concentrations of interleukin-6 and interleukin-10, but down-regulated concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-2, interleukin-8, and interferon-γ in plasma. Greater concentrations of plasma immunoglobulin G were also detected in the GTP group. Overall, the data suggested that GTP supplementation from 15 d prepartum to 30 d postpartum improved the milk yield and health status in cows with hyperketonemia during early lactation.     

地温空调井抽回灌效果影响关键因子识别

为了探析引起地温空调井抽回灌效果的主要制约因素,通过单相关系数分析、主成分绝对得分的源解析模型、拟合度检验和多元线性回归的方法分析影响单井回灌效率的影响因素。结果表明:井径、滤水管孔隙率及抽回灌井数比与单井回灌效率的相关性和拟合度较好,故将井径、滤水管孔隙率及抽回灌井数比作为影响单井回灌效率的关键因子,同时分析了单井回灌效率与关键因子的之间的拟合方程及多元线性回归方程,得到了单井回灌效率与关键因子之间的定量关系。     

甜瓜黄斑病毒SYBR Green Ⅰ实时荧光定量PCR方法

为建立检测甜瓜黄斑病毒(melon yellow spot virus, MYSV)的SYBR Green Ⅰ实时荧光定量PCR(qPCR)方法。基于MYSV核衣壳蛋白基因保守序列设计qPCR特异性引物对,针对引物退火温度、引物浓度、特异性和敏感性进行系列优化。结果显示,优化后的qPCR方法最适退火温度为61.3℃,最适引物浓度为0.65μmol·L^-1,特异性强,灵敏度高,比PCR高100倍。以携带目的基因片段的重组质粒为标准品,构建的qPCR标准曲线循环阈值与模板浓度呈良好的线性关系,相关系数为0.999 7。实验样品验证表明建立的qPCR方法可用于MYSV的定量检测。     

氮肥深施对旱作覆膜农田土壤养分含量及春玉米产量的影响

以春玉米为供试作物,于2021—2022年在甘肃省定西市进行不同深度施肥田间试验,以双垄沟覆膜种植为基础,设置3个氮肥施用深度：5 cm(D5)、15 cm(D15)和25 cm(D25),以不施氮(N0)处理为对照,共4个处理,分析不同氮肥施用深度对农田土壤养分含量、干物质积累、水分利用及玉米产量的影响,为优化施肥模式和进一步提升其节水增产效能提供理论依据。结果表明,随施肥深度的增加,农田0～40 cm土层土壤养分含量逐渐增大,D25处理较D15和D5处理的土壤有机碳、全氮、速效磷和速效钾含量分别平均提高8.67%、2.23%、22.11%和15.18%;氮肥深施可提高生育后期养分供给能力,玉米干物质积累量显著提高,D25处理播后105 d～收获期干物质积累量分别较D15和D5提高6.53%和25.30%;2个试验年份农田耗水量和水分利用效率均随着施肥深度的增加逐渐增大,D25处理分别较D15和D5平均提高10.09%和10.08%;D25处理玉米生物产量和经济产量较D5处理分别平均提高27.20%和34.93%。在西北旱作区,氮肥25 cm深施模式能为玉米提供更好的养分供给,是提高...     

群井开采条件下承压含水层渗流动态模拟

根据有限差分方法 ,编制了承压含水层渗流模拟软件。对某水源地在群井开采情况下地下水渗流动态进行数值模拟。模拟结果显示 ,位于高渗区的井 ,压降漏斗较其它井大 ,压力下降快。当地层中存在裂隙时 ,平行裂隙方向压力下降比垂直裂隙方向要快得多。抽水初期 ,地层压力下降很快 ,以后 ,压力下降逐渐变缓 ,位于低渗透区的井逐渐不能满足流量要求。因此做好地下水资源规划十分重要     

Development of House Architectural Area Computing and Plotting System

The exact computing of house architectural area and apportionment of area is strongly related to the actual interests of people. If it is done manually,there are many problem such as longer time spending, more workload, requiring detailed verification, nonstandard calculating report. Using ActiveX automation and ADO techniques, the house architectural area computing and plotting system was developed under Autocad R14 environment with VB 6.0 and Access database in order to solve above problem. The developed system can automatically compute house architectural area and apportionment of area for each floor of each building and can print the related data report and drawing. The system works well in many departments of house measurement. Several key technical problem of the system are discussed.