Safety of an attenuated West Nile virus vaccine, live Flavivirus chimera in horses

REASON FOR PERFORMING STUDY: West Nile virus (WNV) infection is endemic and able to cause disease in naive hosts. It is necessary therefore to evaluate the safety of new vaccines. OBJECTIVES: To establish: 1) the safety of a modified live Flavivirus/West Nile virus (WN-FV) chimera by administration of an overdose and testing for shed of vaccine virus and spread to uninoculated sentinel horses; 2) that this vaccine did not become pathogenic once passaged in horses; and 3) vaccine safety under field conditions. METHODS: There were 3 protocols: 1) In the overdose/shed and spread study, horses were vaccinated with a 100x immunogenicity overdose of WN-FV chimera vaccine and housed with sentinel horses. 2) A reversion to virulence study, where horses were vaccinated with a 20x immunogenicity overdose of WN-FV chimera vaccine. Horses in both studies were evaluated for abnormal health conditions and samples obtained to detect virus, seroconversion and dissemination into tissues. 3) In a field safety test 919 healthy horses of various ages, breeds and sex were used. RESULTS: Vaccination did not result in site or systemic reactions in either experimental or field-injected horses. There was no shed of vaccine virus, no detection of vaccine virus into tissue and no reversion to virulence with passage. CONCLUSIONS: WN-FV chimera vaccine is safe to use in horses with no evidence of ill effects from very high doses of vaccine. There was no evidence of reversion to virulence. In addition, administration of this vaccine to several hundred horses that may have been previously exposed to WNV or WNV vaccine resulted in no untoward reactions. POTENTIAL RELEVANCE: These studies establish that this live attenuated Flavivirus chimera is safe to use for immunoprophylaxis against WNV disease in horses.     

Efficacy, duration, and onset of immunogenicity of a West Nile virus vaccine, live Flavivirus chimera, in horses with a clinical disease challenge model

REASON FOR PERFORMING STUDY: West Nile virus (WNF) is a Flavivirus responsible for a life-threatening neurological disease in man and horses. Development of improved vaccines against Flavivirus infections is therefore important. OBJECTIVES: To establish that a single immunogenicity dose of live Flavivirus chimera (WN-FV) vaccine protects horses from the disease and it induces a protective immune response, and to determine the duration of the protective immunity. METHODS: Clinical signs were compared between vaccinated (VACC) and control (CTRL) horses after an intrathecal WNV challenge given at 10 or 28 days, or 12 months post vaccination. RESULTS: Challenge of horses in the immunogenicity study at Day 28 post vaccination resulted in severe clinical signs of WNV infection in 10/10 control (CTRL) compared to 1/20 vaccinated (VACC) horses (P<0.01). None of the VACC horses developed viraemia and minimal histopathology was noted. Duration of immunity (DPI) was established at 12 months post vaccination. Eight of 10 CTRL exhibited severe clinical signs of infection compared to 1 of 9 VACC horses (P<0.05). There was a significant reduction in the occurrence of viraemia and histopathology lesion in VACC horses relative to CTRL horses. Horses challenged at Day 10 post vaccination experienced moderate or severe clinical signs of WNV infection in 3/3 CTRL compared to 5/6 VACC horses (P<0.05). CONCLUSIONS: This novel WN-FV chimera vaccine generates a protective immune response to WNV infection in horses that is demonstrated 10 days after a single vaccination and lasts for up to one year. POTENTIAL RELEVANCE: This is the first USDA licensed equine WNV vaccine to utilise a severe challenge model that produces the same WNV disease observed under field conditions to obtain a label claim for prevention of viraemia and aid in the prevention of WNV disease and encephalitis with a duration of immunity of 12 months.     

Characterization,phylogenetic analyses and pathogenicity of Bipolaris setariae associated with brown stripe disease on sugarcane in Yunnan,China

Brown stripe disease is a severe foliar fungal disease of sugarcane worldwide and is widespread in all sugarcane planting areas in China. Brown stripe is a major disease that seriously affects the output and quality of the sugarcane industry in Yunnan Province, China's second-largest sugar base, while the pathogen of this disease remains not yet fully understood. To address this, we isolated and identified the fungi associated with 68 leaf samples showing typical symptoms of brown stripe from 22 sugarcane varieties in different areas of Yunnan Province. A total of 113 isolates were obtained, which were morphologically similar. Of these, 64 representative isolates were sequenced for the internal transcribed spacer region (ITS), GAPDH and EF-1α loci. All representative isolates grouped with the type strain of Bipolaris setariae in the phylogenetic trees inferred with individual and concatenated sequences of ITS, GAPDH and EF1-α. Pathogenicity test results showed that B. setariae strains were able to induce typical symptoms of brown stripe. The results obtained in this study clarify that only B. setariae is associated with sugarcane brown stripe in Yunnan, China. It is recorded here for the first time as a pathogen causing sugarcane brown stripe in Yunnan, and it is able to infect many major cultivars and new varieties, posing a new threat to the sugar industry in Yunnan Province. In addition, these results provide the scientific basis for the future breeding of disease-resistant varieties and effective prevention and control of sugarcane brown stripe disease.     

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。     

吡蚜酮在马铃薯中的残留与膳食风险评估

为评价马铃薯中吡蚜酮残留引发的膳食风险。2016年-2018年开展了3年12地规范残留试验,构建吡蚜酮在马铃薯中的残留检测方法,并对我国一般人群进行了膳食风险评估。马铃薯样品使用2%乙酸乙腈提取,C18净化,外标法定量,UPLC-MS/MS检测。在0.01～1 mg/kg添加水平下,吡蚜酮在马铃薯中的平均回收率为75%～85%,相对标准偏差为3%～8%。12地最终残留试验表明,50%吡蚜酮水分散粒剂按有效成分225 g/hm²施药2次,最后1次施药后14、21 d马铃薯中吡蚜酮残留量分别≤0.018 mg/kg和≤0.012 mg/kg。膳食风险评估表明,吡蚜酮的国家估算每日摄入量(NEDI)为1.633 9 mg/kg,风险商(RQ)为86.4%。马铃薯中吡蚜酮残留不会对一般人群健康造成不可接受的风险。     

超高效液相色谱-串联质谱法测定柑橘中的草甘膦及其代谢物残留

本研究首次建立了一种超高效液相色谱-串联质谱法(UHPLC-MS/MS)同时测定柑橘中草甘膦及其代谢物N-乙酰草甘膦?氨甲基磷酸(AMPA)和N-乙酰AMPA?样品经过0.5%甲酸水提取, 以十八烷基硅烷键合硅胶柱(C18)净化, 利用水和200 mmol/L碳酸氢铵溶液(含0.1%氨水)作为流动相梯度洗脱, 在多反应监测模式下定量分析?结果表明, 草甘膦及其代谢物在0.05~1 mg/kg范围内线性关系良好, 相关系数均大于0.99; 草甘膦及其3个代谢物在柑橘全果和果肉基质中3个加标水平下回收率为 70.5%~109.5%, 相对标准偏差(RSD)为0.6%~10.1%, 定量限为0.05 mg/kg?本方法的前处理样品无需衍生, 简便?快捷?高效?准确可靠, 可用于柑橘中草甘膦及其代谢物N-乙酰草甘膦?AMPA和N-乙酰AMPA的定量检测?     

机动车废气分析仪信号处理系统的智能化设计

提出了机动车废气分析仪的智能化组成及其实现信号处理的全过程。详细阐述了信号处理的软硬件的实现。实验结果表明：本系统克服了传统不分光红外线分析仪固有的缺陷。进一步保证了测量数据的正确性和准确性。     

棉花γ辐射诱变后代M5农艺经济性状的遗传变异

利用250Gy的Co60γ射线对棉花3个品种的干种子进行辐射处理,对其M5农艺经济性状的遗传变异进行分析。结果表明:3个品种的M5群体的铃重、单株铃数、株高、果枝数和麦克隆值的平均变异系数均超过10.0%以上,3个品种的辐射诱变后代M5群体表型性状变异系数存在明显差异;3个品种的M5群体的铃重与2.5%跨长和比强度均呈显著或极显著正相关,在其它性状间的相关性上3个品种的M5群体间存在较大差异;3个品种的M5群体的2.5%跨长均为第一主成分的主要因子。这阐明了辐射对不同棉花品种的诱变效果存在差异,揭示了辐射创造了丰富的遗传变异。     

遮光对彩色棉的色泽及纤维品质的影响

通过对棕色棉和绿色棉不同发育时期棉铃的遮光处理,研究了遮光对彩色棉的纤维色泽和纤维品质的影响。结果表明,棕色棉和绿色棉在-5~55 DPA(开花后的天数)遮光后,其纤维还表现各自的纤维色泽,但其纤维色泽鲜艳度和稳定性在不同遮光时期有一定程度的差别。棕色棉在25DPA以前遮光的纤维颜色比对照变淡;绿色棉在不同生育期遮光后颜色比对照深。遮光对彩色棉和白色棉纤维品质的若干指标有一定的影响,在5~15DPA遮光的纤维都比不遮光的对照粗,在30DPA遮光的纤维长度和强度都较好,35DPA以后遮光的纤维长度和强度随遮光时间的延长越来越差,在45~50DPA套袋遮光后的纤维长度、强度和整齐度都比不遮光的要低得多。     

湿地松优良半同胞家系蛋白质及糖类对水分胁迫的生理响应

为研究湿地松优良半同胞家系蛋白质及糖类对水分逆境的生理响应，采用盆栽试验和吸光度测定结合的方法，以湿地松普通种子为对照（CK），测定了七个湿地松半同胞家系在正常水分状态（土壤相对含水量为70%）、弱度水分胁迫（土壤相对含水量为55%~60%）、中度水分胁迫（土壤相对含水量为35%~40%）和强度水分胁迫（土壤相对含水量为20%~25%）条件下的可溶性蛋白质、还原性糖、可溶性糖的含量，研究结果表明：无论是不同水分梯度还是不同家系水平，蛋白质、还原性糖、可溶性糖含量的差异均达极显著性水平。随着水分胁迫程度的加剧，各家系蛋白质含量呈先升后降的趋势，在水分胁迫初期，各家系蛋白质含量增加，水分胁迫中、后期各家系蛋白含量呈直线下降；随着水分胁迫的加剧，各家系还原性糖、可溶性糖含量呈直线上升。家系1027、101、464在水分胁迫条件下具有较高的可溶性糖含量，其抗旱性较其它家系要强，是这三个家系速生、高产的生理原因之一。