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.     

A comparison of the effects of carbon dioxide and medical air for abdominal insufflation on respiratory parameters in xylazine-sedated sheep undergoing laparoscopic artificial insemination

AIMS: To determine if abdominal insufflation with medical air will improve oxygenation and ventilation parameters when compared to insufflation with CO₂ in xylazine-sedated sheep undergoing laparoscopic artificial insemination (AI).

METHODS: Forty-seven sheep underwent oestrus synchronisation and were fasted for 24 hours prior to laparoscopic AI. Each animal was randomised to receive either CO₂ or medical air for abdominal insufflation. An auricular arterial catheter was placed and utilised for serial blood sampling. Respiratory rates (RR) and arterial blood samples were collected at baseline, after xylazine (0.1?mg/kg I/V) sedation, 2 minutes after Trendelenburg positioning, 5 minutes after abdominal insufflation, and 10 minutes after being returned to a standing position. Blood samples were collected in heparinised syringes, stored on ice, and analysed for arterial pH, partial pressure of arterial O₂ (PaO₂), and CO₂ (PaCO₂). The number of ewes conceiving to AI was also determined.

RESULTS: Repeated measures ANOVA demonstrated temporal effects on RR, PaO₂, PaCO₂ and arterial pH during the laparoscopic AI procedure (p<0.001), but no difference between insufflation groups (p>0.01). No sheep experienced hypercapnia (PaCO₂>50?mmHg) or acidaemia (pH<7.35). Hypoxaemia (PaO₂<70?mmHg) was diagnosed during the procedure in 14/22 (64%) ewes in the CO₂ group compared with 8/23 (35%) ewes in the medical air group (p=0.053). Overall, 15/20 (75%) ewes in the CO₂ group conceived to AI compared with 16/22 (72.7%) in the medical air group (p=0.867).

CONCLUSIONS AND CLINICAL RELEVANCE: There were no statistical or clinical differences in RR, PaO₂, PaCO₂, pH, or conception to AI when comparing the effects of CO₂ and medical air as abdominal insufflation gases. None of the sheep experienced hypercapnia or acidaemic, yet 42% (19/45) of sheep developed clinical hypoxaemia, with a higher percentage of ewes in the CO₂ group developing hypoxaemia than in the medical air group. Based on the overall analysis, medical air could be utilised as a comparable alternative for abdominal insufflation during laparoscopic AI procedures.     

桃蚜电压门控钠离子通道基因cDNA克隆及其生物信息学分析

克隆获得桃蚜电压门控钠离子通道基因cDNA序列，明确钠离子通道的典型特征，为研究桃蚜抗性分子机理奠定基础。采用实验技术主要有RT-PCR和PCR，克隆桃蚜钠离子通道基因cDNA序列，利用相关软件对其序列进行生物信息学分析。克隆得到两段cDNA序列MpNa_v-1（NCBI登录号：MN124170）和MpNa_v-2（NCBI登录号：MN176136）。MpNa_v-1长度为2945 bp，包括2877 bp的完整开放阅读框，共编码958个氨基酸；MpNa_v-2长度为3546 bp，包括3486 bp的完整开放阅读框，共编码1161个氨基酸。MpNa_v-1和MpNa_v-2共同组成桃蚜的钠离子通道α亚基，MpNa_v-1包含同源结构域Ⅰ和同源结构域Ⅱ，MpNa_v-2包含同源结构域Ⅲ和同源结构域Ⅳ。同源比对发现，桃蚜与豌豆蚜和高粱蚜钠离子通道基因相似度分别高达97.67%和97.65%，所克隆序列包含昆虫钠离子通道α亚基典型特征，具有MFM模块，并含有蚜虫类钠通道特有模块DENS。成功地克隆桃蚜钠离子通道基因，为阐明其对拟除虫菊酯类药剂产生靶标抗性的分子机制奠定基础。     

橡胶园酸性、中性土壤交换性钙、镁测定方法研究

为了获得简单、易操作、准确度高的橡胶园酸性、中性土壤交换性钙、镁测定方法,本试验通过响应面优化试验,建立称样量、提取液体积、提取方式与交换性钙测定结果之间的数学模型。结果表明：在25℃条件下,以1 mol/L的乙酸铵为土壤浸提液,称样量为2 g、提取液体积175 mL、振荡36 min,为橡胶园酸性、中性土壤的最佳提取条件。将上述条件下得到的结果与标准方法条件下得到的结果相对比,两结果的相对误差在1.25~3.04之间,均属合理范围。利用标准样品对优化方法进行试验验证,结果与推荐值相符。改进后的方法分析成本低、操作简便、测定结果准确、稳定性好,可用于橡胶园酸性、中性土壤交换性钙、镁的测定。     

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.     

超高效液相色谱—串联质谱法检测鸡血液样品中喹诺酮类药物残留

旨在建立鸡血液样品中多种喹诺酮类药物残留的超高效液相色谱—串联质谱方法。样品经乙腈提取,经Waters Acquity UPLC BEH C₁₈色谱柱分离,以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱,电喷雾正离子（ESI⁺）模式电离,MRM扫描模式检测。经方法学验证,该方法的线性关系、回收率、精密度均符合要求。应用超高效液相色谱—串联质谱法建立了鸡血液样品中多种喹诺酮类药物同时检测的方法,实现了动物血液样品中12种喹诺酮类药物同时定性、定量分析。     

秀珍菇原基形成相关基因PpFBD1的克隆与表达研究

前期研究中通过对秀珍菇经低温诱导后不同发育阶段样本进行转录组测序及差异表达基因分析发现,ID为Cluster-6377.64510的基因(命名为PpFBD1)在原基形成前期的样本中高表达。为进一步了解分析其表达特性及功能,根据转录组测序结果设计特异性引物,利用RT-PCR技术从秀珍菇中克隆获得了该基因的cDNA全长。该基因由342个核苷酸组成,编码一个由113个氨基酸组成的蛋白,蛋白相对分子质量约11 ku。进化树分析显示,PpFBD1编码蛋白与糙皮侧耳hydrophobin1编码的疏水蛋白亲缘关系最近。Gene Ontology功能分析表明,PpFBD1主要参与真菌类细胞壁的合成。荧光定量RT-PCR检测显示,该基因在菌丝体经低温诱导后恢复至室温阶段(原基形成前期)表达量最高,这表明其可能参与调控秀珍菇原基形成过程。本研究结果为阐明秀珍菇原基形成机制提供参考。     

中药对肉仔鸡脂质代谢及肉品质的影响

将120只1周龄肉仔鸡随机分成4组,每组30只,前3组(试验Ⅰ、Ⅱ、Ⅲ组)分别喂以含1%的3种不同中药组方的饲粮,第Ⅳ组为对照组,只喂基础日粮.试验期6周.结果(1)与对照组相比,Ⅰ组血清TC、LDL-C极显著下降(P＜0.01);Ⅱ组血清TC、LDL-C极显著下降(P＜0.01),TG也下降(P＜0.05),HDL-C显著升高(P＜0.01);Ⅲ组血清LDL-C极显著下降(P＜0.01),HDL-C则显著升高(P＜0.01);试验Ⅰ～Ⅲ组血清总脂都极显著下降(P＜0.01);与Ⅱ组相比,Ⅰ组血清LDL-C、Ⅲ组TC存在显著差异(P＜0.01).(2)与对照组相比,胸肌组织中,各试验组TG极显著下降(P＜0.01),Ⅱ组TC显著下降(P＜0.05);腿肌中,各组TC显著下降(P＜0.05),TG下降极显著(P＜0.01);肝脏中各组TC、TG均极显著下降(P＜0.01);与Ⅱ组相比,Ⅰ组肝TG差异极显著(P＜0.01);Ⅲ组胸肌TG、肝TG有显著差异性(P＜0.05).(3)7周龄末鸡体重与对照组相比,Ⅱ组差异极显著(P＜0.01),Ⅲ组差异显著(P＜0.05);Ⅱ组料肉比比对照组极显著下降(P＜0.01).(4)肌肉中的营养成分,Ⅰ、Ⅱ组腿肌中粗脂肪比对照组极显著下降(P＜0.01),Ⅱ组腿肌中粗蛋白含量显著升高(P＜0.01).结论中药组方能不同程度地调整鸡脂质代谢,提高生产性能和改善鸡肉品质.在3个不同中药组方中,综合效果Ⅱ方最优.     

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。     

海南岛昌江县昌化镇草地及其利用途径

海南岛昌江县昌化镇草地,用点样法取得数据,应用spss软件聚类分析,可划分为五类。以和它草——三芒草群落面积最大,最有代表性;金须茅——蔓荆子——绢毛飘拂草则局限于滨海沙地上。文章末尾就利用问题作了讨论。