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。     

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.     

The Use of a Metabolic Disorder Index as a Predictor for Metabolic Eliminations in Endurance Horses

The equine endurance race involves both aerobic and anaerobic metabolisms of the horse. The intense physical activity over an extended period often causes susceptible horses to develop metabolic signs or problems resulting in elimination from races. Thus, the objective of this study was to develop a method for prediction and validation of a metabolic disorder index (MDI) to be used before endurance races. Three hundred seventy-five Arabian (n = 152) and Arabian cross (n = 223) endurance horses aged from 6 to 15 years and weighing between 350 and 450 kg were selected for the study in Malaysia. Blood samples were collected at pre- and post-race periods. The significant (P < .05) findings in horses with metabolic disorder were packed cell volume (0.50 ± 0.06 LL−1), creatine kinase (1,275.89 ± 121.45 UL−1), interleukin-6 (2.01 ± 0.89 ng/mL), decreased glutathione reductase (26.57 ± 3.95 ng/mL), and chloride (94.98 ± 8.12 mmol/L). A new method called MDI was developed as a predictor for horses with the potential to develop metabolic disorders in endurance races. The MDI indicated a higher value greater than 5.5 for those eliminated and lower value below 5.5 for those that completed the race successfully, this proved to be accurate in the prediction of metabolic disorder in endurance horses. The MDI is an innovative and simple method used as a prediction method that will assist the equine endurance society to reduce the rate of elimination and to safeguard against serious medical problems during endurance races in the tropics.     

The microbial diversity and structure in peatland forest in Indonesia

The microbial community structure and function under forest in tropical peatlands are poorly understood. In this study, we investigated the microbial community structure and diversity in natural peat swamp forest soil, disturbed peat soil and mineral soil in Central Kalimantan, Indonesia, using 454 pyrosequencing. The results showed that the natural peat soil had the greatest fungal species richness (Chao1), which was significantly (p < .05) larger than that in the other two soils. Community structure of both fungi and bacteria in natural peat soil differed significantly from that in the disturbed peat soil (p = .039 and p = .045, respectively). Ascomycota (40.5%) was the most abundant phylum across the three soils followed by Basidiomycota (18.8%), Zygomycota (<0.1%) and Glomeromycota (<0.1%). The linear discriminant analysis with effect size (LEfSe) showed that Ascomycota (p < .05) and genus Gliocephalotrichum (p < .05) dominated in natural peat soil. Functionally, pathotrophs were more abundant in disturbed peat soil (p < .05). Proteobacteria (43.8%) were the most abundant phylum followed by Acidobacteria (32.6%), Actinobacteria (9.8%), Planctomycetes (1.7%). Methylocystis, Telmatospirillum, Syntrophobacter, Sorangium and Opitutus were the more abundant genera in disturbed peat soil, whereas Nevskia and Schlesneria were more abundant in mineral soil and natural peat soil, respectively. The natural peat forest soil supported a more diverse microbiology; however, the land use of such a soil can change its microbial community structure. The results provide evidence that the disturbance of tropical peat land could lead to the introduction and spread of a large number of fungal diseases     

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.     

富锗酵母培养物对下笼蛋鸡肉品质、肠道菌群及结构的影响

本试验旨在研究富锗酵母培养物对下笼蛋鸡肉品质及肠道的影响。选取505日龄健康的海兰褐壳蛋鸡480只,随机分为4组,每组5个重复,每个重复24只鸡。对照组饲喂基础日粮,试验组在基础日粮中分别添加12、16、20 mg/kg的富锗酵母菌培养物。预试期7 d,正试期为42 d。结果表明:与对照组相比,12、16、20 mg/kg组腿肉的蒸煮损失分别降低45.2%、33.6%、39.7%(P<0.01),pH均高于对照组(P<0.01);16mg/kg组腿肌的亮度(L~*)低于对照组(P<0.05);12、16、20 mg/kg组腿肌的剪切力分别比对照组低44.13%、56.07%、50.60%(P<0.01);各试验组胸肌pH均高于对照组(P<0.01),16mg/kg组的pH高于12、20mg/kg组(P<0.05);与对照组相比,16mg/kg组乳酸菌数量升高(P<0.05);空肠中,各试验组的绒毛长度高于对照组(P<0.01);16、20 mg/kg组绒毛长度/隐窝深度高于对照组(P<0.01)。结果显示,日粮中添加富锗酵母培养物可以改善下笼蛋鸡小肠的形态结构和肠道菌群平衡,提高鸡肉品质,建议添加量为16 mg/kg。     

施用锌肥和遮阴互作对花生生长发育、抗病性及产量的影响

采用大田裂区试验，研究了施用锌肥和不同遮阴程度互作对花生生长发育、抗病性及产量的影响。结果表明，与不施锌肥相比，施用锌肥能提高花生不同部位锌含量、增加叶片SPAD值，提高花生叶片中可溶性糖、蛋白质和生长素含量，减少花生病害的发生，平均降低7.1个百分点，花生产量平均增加19.4%。相同施锌水平下，随着遮阴程度的增加，花生不同部位锌的含量和不同生育期叶片SPAD值以及花生叶片中可溶性糖、蛋白质和生长素含量呈增加趋势，花生的发病率比不遮阴对照增加4.8、10.2个百分点，花生产量平均降低16.5%、10.0%。在30%、70%的遮阴条件下，施用锌肥的花生产量比不施锌的分别提高21.1%、25.0%。本试验条件下，施用七水硫酸锌30 kg/hm2，使花生具有较强的抗低温寡照能力及抗病性能，增产显著，可在花生产区推广应用。     

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。     

Spatiotemporal patterns of mountain whitefish (Prosopium williamsoni) in response to a restoration of longitudinal connectivity

To examine the role of longitudinal connectivity on the spatial and temporal dynamics of mountain whitefish (Prosopium williamsoni), we quantified movement and population dynamics following installation of the Landsburg Dam fishway, Cedar River, WA, USA. Mountain whitefish is widely distributed, poorly studied and not the focus of restoration. Before the fishway, mountain whitefish were not observed above the dam. Here, we focus on snorkel counts collected at reach and mesohabitat (e.g. pools) scales over 11 summers on the 20‐km above‐dam segment following restoration. A camera within the ladder provided number, size and movement timing, thereby informing on behaviour and recolonisation. Segment‐scale abundance increased following fish passage reaching an asymptote in 7 years, and mountain whitefish were detected throughout the main stem in 10 years. Annual movement through the ladder increased over time and was positively correlated with instream abundance and discharge, but negatively correlated with water temperature. About 60% of fish movements occurred in spring and early summer, potentially for foraging opportunities. Reach‐scale abundance peaked between 7 and 10 km from the dam; deep, cool (~10.6 to 11.6°C) conditions characterised these reaches. At the mesohabitat scale, mountain whitefish detection increased with depth and velocity after accounting for distance from the dam. Our results show how restoring longitudinal connectivity allowed this nontarget species to colonise newly available habitat. Their response supports the critical roles of longitudinal connectivity and environmental conditions, that manifest at different spatial scales, in dictating how freshwater fish respond to habitat disturbance.