恒温隔绝式荧光PCR检测非洲猪瘟病毒核酸方法的建立

恒温隔绝式荧光PCR(iiPCR)是一种简便、快速及可使用于现场检测的荧光PCR方法.本研究采用农业农村部推荐的检测非洲猪瘟病毒(African swine fever virus,ASFV)核酸的荧光定量PCR方法的引物和探针,通过对引物、探针、Taq酶等反应体系的优化,建立了基于恒温隔绝式荧光PCR的ASFV检测方...     

稻草还田模式对稻田土壤速效氮、钾含量及晚稻生长的影响

设计了高桩翻耕抛栽、覆盖免耕抛栽和高桩免耕抛栽3种模式的田间小区试验。结果表明, 不同稻草还田模式对土壤NH₄⁺-N含量影响较大, 对土壤速效钾含量和土壤NO₃^--N含量影响不大; 高桩翻耕模式的土壤NH₄⁺-N、速效钾含量平均值比高桩免耕和覆盖免耕模式分别提高40.8%、7.5%和36.7%、2.0%; 高桩翻耕能促进水稻群体生长, 增加叶片叶绿素含量, 充分利用光能, 增强光合作用; 稻草还田以高桩翻耕模式的增产效果最好, 比高桩免耕、覆盖免耕分别增产5.8%和9.0%。以高桩翻耕稻草还田模式对土壤速效氮、钾含量及晚稻产量的影响最好。     

优氯净对苏云金杆菌伴孢晶体消毒效果及机理的研究

苏云金杆菌(Bacillus thuringiensis,简称BT菌)的伴孢晶体被优氯净液处理后,在相差显微镜下观察,失去暗蓝色光泽,呈暗黑色,难溶于碱性缓冲液。虽能溶于昆虫的胃液,却失去致病性。用扫描电镜和SDS—聚丙烯酰胺凝胶电泳进一步研究,结果表明:优氯净能破坏伴孢晶体蛋白的一级和立体结构,使晶体蛋白失活,达到消毒的目的。消毒能力测定结果表明:1毫升0.03％有效氯优氯净液在3分钟内能消毒50微升BT菌液(3μg伴孢晶体/μl菌液)。消毒BT菌污染桑叶,用0.01％有效氯优氯净液浸泡3分钟即可。考虑到生产上的安全性,我们建议用0.05％有效氯优氯净消毒3—5分钟。     

Study on the Virulence Gene Distribution and Genetic Evolution of Cattle Shiga Toxin-producing Escherichia coli Isolates

This study was aimed to understand the relationship of virulence gene distribution and genetic evolution between cattle originated Shiga toxin-producing Escherichia coli (STEC) and human originated enterohaemorrhagic Escherichia coli (EHEC) O157. This experiment collected 18 strains STEC in a dairy farm from Jiangsu province and 9 STEC reference strains (human, sheep, swine and avian), according to the method of U.S. Centers for Disease Prevention and Control Center (PulseNet), using the XbaⅠ enzyme digestion and pulsed field gel electrophoresis (PFGE) analysis, virulence genes were detected in some STEC isolates. The virulence gene distribution of O157 from different origin was remarkably different. The cattle originated STEC O157 and the human originated EHEC O157:H7 (EDL933W) had the most similar virulence gene distribution. In contrast, virulence genes were lack in cattle STEC O18 and O26, even though the cattle STEC O18 and O26 had the similar genotype as human EHEC O157:H7 (EDL933W). PFGE of Xba Ⅰ digested chromosomal DNA from 27 isolates of STEC exhibited 22 profiles. In general,the Dice coefficients of different originated STEC ranged from 72% to 100%.Cattle STEC O157 had a high similarity with two strains of human originated EHEC O157, while a low similarity was demonstrated between cattle STEC O157 and STEC O157 of swine and avian. The Dice coefficients of the cattle STEC O157 and the two strains of human EHEC O157 ranged from 83% to 95%. The Dice coefficients of cattle STEC O26 (Ⅶ,Ⅷ) and the two strains of human EHEC O157 were more than 82%. Therefore, it was concluded that the cattle STEC O157 and human EHEC O157 had a closer relationship in terms of virulence gene distribution and in genetic evolution.     

Response to lipopolysaccharide in Octodon degus pups produces age‐related sickness behavior but does not have effects in juveniles

During vertebrate development, the immune function is inefficient and is mainly controlled by innate defense. While there have been detailed studies of various aspects of innate immune function, the effects of this function in the growth of vertebrates is still not well known. Similarly, there is little information regarding how early endotoxin exposure would affect juvenile phenotypes, specifically in a non‐model mammal like a precocial rodent. We evaluated the response to an antigen and its cost in offspring of the rodent Octodon degus. We inoculated pups at 4 different ages (8, 15, 22 and 30 days after birth) with an antigen to determine the ontogeny and costs of the response to an endotoxin. We assessed changes in body mass, body temperature, sickness behavior and the levels of a key mediator of the inflammatory response, the cytokine interleukin‐1β. We also determined the effects of early endotoxin exposure on the resting metabolic rate of juvenile animals (i.e. 90 days after birth). The cytokine levels, body mass and body temperature were unaffected by time of inoculation and treatment. However, pups subjected to inoculation at 22 days after birth with the antigen showed reduced locomotion. Juvenile resting metabolic rate was not affected by early endotoxin exposure. These results suggest that the magnitude of O. degus responses would not change with age. We discuss whether the lack of effect of the response on body mass or body condition is caused by environmental variables or by the precocial characteristics of O. degus.     

ROUTE DEVELOPMENT,ANTIVIRAL STUDIES,FIELD EVALUATION AND TOXICITY OF AN ANTIVIRAL PLANT PROTECTANT NK0238

• Developed a two-step synthetic route to anti-plant-virus candidate NK0238. • NK0238 exhibited a broad antivirus spectrum in greenhouse. • NK0238 showed comparable antivirus activities as controls in field trials. • NK0238 was safe to birds, fish, bees and silkworms. • NK0238 has a very good prospect in commercial development. It has previously been shown that tryptophan, the biosynthesis precursor of Peganum harmala alkaloids, and its derivatives have anti-TMV activity both in vitro and in vivo. Further exploration of this led to the identification of NK0238 as a highly effective agent for the prevention and control of diseases caused by plant viruses, but the existing routes are unsuitable for its large-scale synthesis. This study optimized a route for two-step synthesis of this virucide candidate via reaction of l-tryptophan with triphosgene to produce l-tryptophan-N-carboxylic anhydride, which then reacts with n-octylamine to give NK0238 at up to 94% yield and nearly 97% HPLC purity. In addition, the route was used for the preparation of NK0238 on a>40 g scale permitting further assessment of its antivirus activity in the greenhouse and field experiments, and toxicity tests. NK0238 exhibited useful antiviral activities against a variety of viruses both in greenhouse and field experiments. The toxicity tests showed that NK0238 was not acutely toxic to birds, fish, honey bees and silkworms. The optimized route provides a solid foundation for its large-scale synthesis and subsequent efficacy and toxicity studies, its excellent activity and safety make NK0238 a promising drug candidate for further development.     

6株羊源乳酸杆菌电转化条件的优化

选取质粒pLEM415-egfp电转化源于陕北白绒山羊瘤胃的乳酸杆菌,研究不同生长时期及电场强度对6株羊源乳酸杆菌电转化效率的影响,并优化其转化条件。在37 ℃培养条件下,培养6株羊源乳酸杆菌,当D_{600 nm} 达到0.4、0.6、0.8、1.0和1.2时,离心收集菌体,制备感受态细胞;在电阻200 Ω、电容25 uF条件下,分别采用1.5、2.0和2.5 kV 3种电场强度电击转化感受态细胞。结果表明,羊源植物乳酸杆菌(Lactobacillus plantarum)、干酪乳酸杆菌(Lactobacillus casei)、罗伊乳酸杆菌(Lactobacillus reuteri)、短乳杆菌(Lactobacillus brevis)、棒状乳酸杆菌(Lactobacillus coryniformis)和弯曲乳酸杆菌(Lactobacillus curvatus)获得最高转化效率的D_{600 nm}和电场强度分别为0.8、1.2、1.0、0.6、0.8、0.4和1.5、1.5、1.5、1.5、1.5、2.0 kV。羊源乳酸杆菌最佳转化条件的确立,为后续乳酸杆菌转基因工程菌的构建及其应用奠定了基础。     

黄土高原不同粮草种植模式土壤碳氮及土壤酶活性

通过在陇中黄土高原半干旱区对苜蓿(Medicago sativa)-作物轮作地进行长期定位试验,探讨不同种植模式对土壤碳氮形态及其相关酶活性的影响。6种种植模式分别为苜蓿-苜蓿、苜蓿-休闲、苜蓿-小麦(Triticum aestivum)、苜蓿-玉米(Zea mays)、苜蓿-马铃薯(Solanum tuberosum)和苜蓿-谷子(Setaria italica)。结果表明,苜蓿-作物种植模式不利于土壤总有机碳的积累,而苜蓿翻耕后保持休闲则可维持较高的有机碳含量;与苜蓿连作相比,苜蓿-作物种植模式的土壤有机碳降低了1.60%~23.11%,全氮含量增加了3.81%~21.83%。不同作物对土壤养分吸收利用状况不同,进而引起土壤酶发生变化。与苜蓿连作相比,苜蓿粮食作物种植模式在降低土壤过氧化氢酶和蛋白酶活性的同时,提高了土壤硝酸还原酶活性;其中土壤过氧化氢酶活性和蛋白酶活性分别降低了5.20%~12.30%和15.03%~43.43%,硝酸还原酶活性提高了1.26%~28.79%。苜蓿连作和苜蓿-粮食作物种植模式间的土壤脲酶活性无显著差异(P0.05),但均高于苜蓿-休闲处理。相关性分析结果表明,土壤脲酶活性与土壤有机碳、全氮含量呈显著正相关(P0.05),可作为衡量土壤肥力的指标。     

高密度二氧化碳对食品中蛋白质结构及其加工特性影响研究进展

非热加工技术已广泛应用于食品加工行业,高密度二氧化碳（dense phase carbon dioxide,DPCD）加工技 术作为一种新型的非热加工技术,因其无毒无害、能耗低、对食品品质影响小等特点,成为国内外食品加工技术研 究的焦点。蛋白质作为食品的重要组成成分,直接影响食品的风味、质构及营养。本文简要介绍DPCD加工技术概 况和特点,总结关于DPCD对蛋白质结构及加工特性影响和DPCD钝酶作用的最新研究进展,展望了DPCD加工技术 的发展前景,为今后非热加工技术应用于食品加工提供重要研究思路。