Isolation and Identification of a Variant Porcine Epidemic Diarrhea Virus and the Immune Efficacy of Its Inactivated Vaccine

This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID₅₀,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×10^8.0 TCID₅₀/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×10^8.0TCID₅₀/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China.     

药用植物半夏细胞悬浮培养和植株再生体系建立研究

为了建立甘肃产药用植物半夏的细胞悬浮培养和植株再生体系,通过对其不同外质体筛选、愈伤组织诱导和悬浮培养的愈伤组织类型、激素水平、起始密度、震荡速率等影响因素以及细胞生长状态、生长曲线和pH值变化等指标的研究.结果表明:以叶柄为外质体,选择"松散型"愈伤组织和2×105个/mL的起始培养密度,在附加2,4-D 2.0 mg/L BA 0.5 mg/L NAA0.5 mg/L的1/2 MS培养基上培养,温度(23±2)℃,转速90 rpm,pH值6.0,黑暗培养为最适悬浮培养体系;经检测细胞生长曲线呈"S"型,培养液pH随着细胞的生长先上升后下降,18 d时细胞基数达到最大值;悬浮继代培养后,通过降低或停止震荡转速,在添加BA 0.5 mg/L NAA0.5 mg/L的MS液体和固体培养基上可诱导分化出芽和根,进一步形成完整的植株.     

苎麻悬浮细胞原生质体培养再生植株

苎麻品种浏阳大叶绿的子叶在含有２，４－Ｄ０．５ｍｇ／Ｌ、ＫＴ０．５ｍｇ／Ｌ的ＭＳＢ固体培养基上，形成愈伤组织。愈伤组织经３￣４次继代培养后作液体振荡培养，产生悬浮细胞系。从悬浮系分离的原生质体，只有以海藻酸钠包埋方式培养在ＫＭ８Ｐ培养基中，５０天左右才能形成肉眼可见的小愈伤组织。该愈伤组织在附加２，４－Ｄ０．２ｍｇ／Ｌ、６－ＢＡ０．１ｍｇ／Ｌ的ＭＳＢ生长培养基上增殖，然后转入附加６－ＢＡ２．０ｍｇ     

夏堇悬浮细胞培养与植株再生研究

以夏堇品种“小丑”的组培苗幼嫩叶片为试材,进行愈伤组织诱导、细胞悬浮培养及植株再生诱导的研究.结果表明:MS+1.0 mg/L 6-BA+0.1 mg/L2,4-D的培养基能够使叶片诱导为生长迅速、质地疏松的愈伤组织.愈伤组织最适宜的悬浮培养条件为:MS+ 1.0 mg/L6-BA+0.1 mg/L 2,4-D液体培养基,20 g/L的起始接种量以及100 r/min的震荡培养.将继代3～4次的细胞悬浮颗粒转到1/2MS+1.0 mg/L 6-BA+0.1 mg/L NAA的固体再生培养基上培养,3周后可以诱导分化出芽,进而获得完整植株.悬浮培养有利于夏堇愈伤组织的再生,悬浮培养对提高以夏堇作为模式植物进行的相关研究具有重要的应用价值.     

超微绿茶粉原料茶加工中的几个技术要点

超微绿茶粉是一种能快速冲饮并可作为食品添加剂的纯天然粉末茶。它能最大限度的保持茶叶原有的各种营养成分、药理成分和原料的天然本色,具有良好的悬浮稳定性等有机特性,可直接应用于茶饮料加工或直接作为速溶型饮料,也可用于各种茶叶食品或添加于各类食品中,以强化其营养功效,并赋予食品天然鲜绿色泽和特有的茶叶风味。超微绿茶粉的原料茶加工和常规的茶叶加工有一定的区别,     

27;吡·福·多悬浮种衣剂的研制

介绍了27;吡·福·多悬浮种衣剂有效成分配比的室内筛选、助剂配方研究、主要技术指标和田间药效等.田间试验表明:27;吡*福*多悬浮种衣剂在试验剂量下对棉花出苗无不良影响,以1∶40～60的药种比例处理棉花种子,对棉花苗期病虫害(地老虎、棉蓟马、立枯病)具有较好的防效.     

转玉米C4型pepc水稻成熟胚为外植体的水稻悬浮细胞系的优化

为了研究外源玉米光合作用关键酶C4型磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因在C3植物水稻中提高光合效率的分子机理,本研究以高表达玉米C4型pepc水稻(PC)和未转基因野生型Kitaake (WT)为材料,探索高效且稳定水稻悬浮细胞系建立的方法.结果表明:PC成熟胚愈伤组织的诱导培养基中添加2 mg/L2,4-D和1 mg/L 6-BA,可出诱导出高频的愈伤组织;再通过3次继代培养(2～0.5 mg/L 2,4-D,2～0.2 mg/L 6-BA,1 mg/L ABA),逐步降低2,4-D和6-BA的浓度,可获得了疏松、颗粒状且分散的愈伤组织块;然后转接到水稻悬浮细胞液体培养基中(0.5 mg/L2,4-D,0.2 mg/L 6-BA和1 mg/LABA),在28℃下培养,新原液的比例为3∶1,经42 d,可建立稳定且高效的供试水稻悬浮细胞系.     

竹子填料海水曝气生物滤器除氮性能

生物滤器是循环水养殖系统的关键水处理单元,主要用于去除水体中水溶性的氮化物.采用人工模拟海水养殖废水,在系统运行的水力停留时间HRT为1h,水温为18～25 ℃,气水比为3∶1,初始C/N＝3∶1,pH为8.05～8.53条件下,对竹子填料浸没式生物滤器的挂膜过程和稳定运行阶段系统去除氨氮的运行特性,以及挂膜过程中的硝化细菌群落变化进行了实验研究.结果表明,在较低的NH+4-N浓度条件下,采用竹子填料的生物滤器有较快的挂膜速度,挂膜成功后滤料表面上生长的氨氧化细菌和亚硝酸氧化细菌的数量分别为4.5×105、1.5×105(光面)和1.1×106 CFU/ml(粗面).具有较高且稳定的氨氮去除效果,氨氮去除效率达到80%,出水浓度小于0.06 mg/L,满足海水循环养殖系统中的应用要求.     

火山岩和沸石填料BAF深度处理印染废水的效能比较

针对纺织印染行业水质不达标回用率低的问题,采用火山岩和沸石为填料的曝气生物滤池对纺织印染废水二级生化出水进行了深度处理,进行了两反应器出水效能的比较。研究表明:火山岩填料深度处理纺织印染废水的效能要高于沸石填料,建议气水比在10∶1左右,出水CODcr为36~48mg/L,出水色度35~55度,NH3-N浓度0.5~1.7mg/L,出水浊度范围0.9~3NTU。