马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株，本研究以EHV-1 XJ2015株DNA为模板，对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析，并扩增TK基因左右重组臂TKL和TKR，构建质粒pUC-TKLR，将扩增后的增强绿色荧光蛋白（EGFP，含有CMV+polyA）插入pUC-TKLR质粒，构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示，XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高，分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%；与EHV-3分离株同源性均最低，分别为72.9%、59.4%和62.1%；遗传进化分析显示，3个基因均与国外EHV-1同属于一个遗传进化分支，与EHV-9和EHV-4进化关系较近，但与EHV-3进化关系较远，表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显，没有明显的地域性特征，功能基因保守且进化缓慢，同源基因功能相同或相近；经PCR扩增、酶切、测序及转染鉴定，本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建，为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。     

同期排卵处理母牛体温和活动量变化规律及效果分析

为探究同期排卵处理母牛体温和活动量变化规律及不同同期排卵技术处理效果，指导同期排卵技术优化。本研究自动监测了18头20月龄左右同期排卵（GnRH-PG-GnRH）处理荷斯坦母牛和17头产后40~60 d预同期排卵（PG-PG-GnRH-PG-GnRH）处理荷斯坦母牛的体温和活动量，应用自动检测系统进行母牛发情监测。结果发现，同期排卵处理母牛发情时阴道温度平均升高（0.43±0.20）℃，持续（12.37±2.73）h；活动量平均升高（18.28±18.61）倍，持续（11.00±1.68）h；排卵时阴道温度平均下降（0.20±0.10）℃，持续（11.00±1.68）h。自动化发情监测显示，同期排卵处理母牛7头发情并排卵；预同期排卵母牛GnRH处理前全部发情排卵。两种同期排卵处理，虽可改变母牛性周期进程，促进母牛性周期同步化，但均难以使母牛性周期完全同步。因此，将同期排卵-定时输精和发情鉴定技术科学结合才能取得更好的繁殖效果。     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

Establishing an eyeball-weight relationship for Litopenaeus vannamei using machine vision technology

This study aimed to establish a shrimp eyeball-weight relationship model for Litopenaeus vannamei using machine vision technology. A total of 295 shrimp were sampled from a recirculating aquaculture system (RAS). The long-axis length (d), body length (L), and body weight (W) of each individual was measured. The long axis length of the shrimp eyeball was identified and measured using machine vision technology. Continuous fitting and piecewise fitting models were used to construct the eyeball-weight relationship model for L. vannamei. The continuous fitting relationship model was described as: W = 38.865d^2.7914, while the piecewise model was described as: d < 2 mm, W = 0.0326d^3.7363, R² = 0.9288; 2 mm ≤ d < 3.9 mm, W = 0.0401d^3.104, R² = 0.9629; 3.9 mm ≤ d < 5.8 mm, W = 0.0421d^3.0311, R² = 0.9216; 5.8 mm < d, W = 0.103d^2.6226, R² = 0.9457. The root mean square error (RMSE) of the piecewise fitting model (0.0244, 0.1575, 0.5034, 0.7072) was smaller than the continuous fitting model (0.8229). The correlation coefficient (R²) of the piecewise model (0.9288, 0.9629, 0.9216, and 0.9457) was similar to that of the continuous fitting model (R² = 0.9621). The results indicated that the piecewise fitting model is suitable for calculating the biomass of L. vannamei in RAS and provides a novel way of estimating the biomass of L. vannamei cultured in RAS. The piecewise fitting model can also provide the foundation of evaluating the production of shrimp using underwater image recognition in intelligent aquaculture systems.     

Measuring trail users’ perception of crowding in a peri-urban nature park: A best-worst scaling experiment

This paper evaluates the perception of crowding, its determinants and relationship with trail users’ experience in a peri-urban nature park. Existing crowding surveys rely heavily on a single-item 9-point crowding scale which was originally developed for backcountry areas. Receiver operating characteristic (ROC) analysis was used to test the validity and reliability of this measure in detecting crowding issues and formulating visitor capacity limits. Low validity and reliability raised concerns about the applicability of this scale in similar frontcountry areas. Inability of this simple crowding measure to detect a visitor threshold brings to the fore its another important limitation – ignorance of the complexity of crowding phenomenon. The perception of crowding varies with various social, psychological and situational factors, and a best-worst scaling (BWS) experiment was designed to simultaneously evaluate different aspects of crowding in the Medvednica Nature Park, a peri-urban nature park near Zagreb, Croatia. The findings of BWS experiment revealed that occupancy of mountain huts, number of other hikers on the trail and amount of litter along the trail on average contributed most to the trail users’ sense of crowding. A latent class analysis identified a large heterogeneity in the perception of crowding; while one group of trail users (63%) was more disturbed by the direct social impacts, the others (37%) were more disturbed by the secondary impacts of other visitors, especially by the negative externalities from road traffic in the park. Older visitors had a greater tendency to primarily associate crowding with the environmental conditions. A higher impact of trail use level in the perception of crowding, younger age and more frequent use were associated with a more negative evaluation of crowding. The park management should closely monitor road traffic in the park and visitor activity on highly used trails, especially on those popular among younger trail users.     

Influence of soil phosphorus status,texture, pH and metal content on the efficacy of amendments to pig slurry in reducing phosphorus losses

Cost‐effective strategies for using chemically amended organic fertilizers need to be developed to minimize nutrient losses in surface and groundwater. Coupling specific soil physical and chemical characteristics with amendment type could increase their effectiveness. This study investigated how water‐extractable phosphorus (P) was affected by chemical amendments added to pig slurry and how this effect varied with soil properties. A 3‐month incubation study was conducted on 18 different mineral soils, stored at 10 °C and 75% humidity and treated with unamended and amended slurry which was incorporated at a rate equivalent to 19 kg total P (TP )/ha. The amendments examined were commercial‐grade liquid alum, applied at a rate of 0.88:1 [Al:TP ], and commercial‐grade liquid poly‐aluminium chloride (PAC ), applied at a rate of 0.72:1 [Al:TP ]. These amendments were previously identified by the authors as being effective in reducing incidental losses of P. The efficacy of the amendments varied with the soil test P, the degree of P saturation (DPS ) and the Mehlich aluminium, iron and calcium, but not soil texture. Chemical amendments were most effective in soils with DPS over approximately 20%. Due to their high cost, the incorporation of amendments into existing management practices can only be justified as part of a holistic management plan where soils have high DPS .     

同源四倍体及其原二倍体黑皮冬瓜光合光响应模型筛选

为了筛选出最适合黑皮冬瓜Benincasa hispida (Thunb.) Cogn.的光合光响应模型,为其育种提供理论依据,以同源四倍体及其原二倍体黑皮冬瓜为试验材料,对8种经典的光合光响应模型适用性进行了比较分析。结果表明：二次多项式能够表现出光抑制情况,但在拟合过程中出现暗呼吸速率为正值、光补偿点为负值及无法解释当光强达到饱和后光合速率快速下降的问题;直角双曲线、非直角双曲线及指数函数Ⅰ、指数函数Ⅱ无法直接求取光饱和点、光补偿点,结合常用的光饱和点的计算方法得到的光饱和点与实测值均存在较大的偏差,且指数函数Ⅱ在计算光饱和点时表现出明显的人为性,也无法拟合光抑制情况,但4种模型拟合得到的光补偿点均与实测值相差不大;指数修正模型因系数β为负值,无法求取四倍体黑皮冬瓜材料的光饱和点和最大净光合速率,且拟合得到的四倍体黑皮冬瓜的光补偿点明显低于实测值;直角双曲线修正模型计算得到的暗呼吸速率及二倍体黑皮冬瓜的光饱和点明显低于实测值,但获得的四倍体及其二倍体的最大净光合速率与实测值最接近,说明其在拟合最大净光合速率上有优势;整体上分段函数计算得到的黑皮冬瓜的各光合参数与实测值最为接近,与实测值的平均相对误差最小,也能很好的拟合发生光抑制部分的光响应曲线。分段函数拟合同源四倍体及其原二倍体黑皮冬瓜光合光响应曲线效果较其他模型效果好,分段函数模型为黑皮冬瓜最适合的光合光响应模型。     

异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。     

6株牛传染性鼻气管炎病毒BICP4基因序列的同源性分析

牛传染性鼻气管炎病毒的立即早期启动基因IER4.2编码感染细胞蛋白BICP4基因。各种疱疹病毒的BICP4基因的Ⅰ，Ⅲ，Ⅴ区变异性较大，并为其特征区。为了比较分离自不同牛种，不同部位的5种病毒株BICP4蛋白I区基因的差异性，本文根据已发表的IBBV K22毒株重复序列IRs中BICP4的基因序列设计了一特异性引物，对各毒株进行PCR扩增，均得到约540bp的片段，将其克隆，测序并连同K22株进行同源性比较。结果表明，6株病毒的核苷酸序列及氨基酸序列同源性均在98%以上，说明这些毒株重复序列中的早期基因高度保守，在一定程度上也显示出IBRV的高度保守性。