1. Vongxay K, Wang SN, Zhang XF, Wu BB, Hu HX, Pan ZJ, Cheng SY, Fang WH. Pathogenetic Characterization of Vibrio parahaemolyticus Isolates from Clinical and Seafood Sources. International Journal of Food Microbiology, 2008, . 王淑娜,方维焕。荧光定量PCR法检测副溶血弧菌tdh基因的表达差异。畜牧与兽医,2008,已录用。3. 陈 阳,吴 迪,陈雪燕,赵焕灿,扈鸿霞,Cheick A Diakite,方维焕。猪链球菌2型分离株对野生斑马鱼的致病性。动物医学进展,2008,已录用。4. 刘刚,李肖梁。犬瘟热病毒H基因的序列分析与表达。畜牧与兽医,2008,已录用。5. 陈健舜, 李肖梁,田国明,马有智,方维焕。检测猪链球菌2型的三重PCR方法建立与应用。中国兽医学报, 2008,已录用。6. Vongxay K, Pan ZJ, Zhang XF, Cheng SY, Mei LL, XU C, Wang SN, Fang WH. Occurrence of pandemic clones of Vibrio parahaemolyticus isolates from seafood and clinical samples in a Chinese coasting province. Foodborne Pathogens and Disease, 2008, 5(2):. Yang ZZ, Shuai JB, Dai XJ, Fang WH. A survey on porcine circovirus type 2 infeciton and phylogenetic analysis of its ORF2 gene in Hangzhou, Zhejiang Province. Journal of Zhejiang Univ Science B, 2008, 9(2):. Jiang LL, Chen JS, Xu JJ, Zhang XF, Wang SN, Zhao HC, Vongxay K, Fang WH. Virulence characterization and genotypic analyses of Listeria monocytogenes isolates from food and processing environments in eastern China. International Journal of Food Microbiology, 2008, 21(1): . Shuai JB, Li XL, Chen N, Chen CY, Fang WH. Characterization and potential use of truncated PCV2 capsid protein and its polyclonal antibody for diagnosis of PCV2 infections. Acta Microbiologica Sinica, 2008:4891):. Shuai JB, Wei W, Jiang LL, Li XL, Chen N, Zhang ZF, Fang WH. Mapping of nuclear localization signals in open reading frame 2 protein from porcine circovirus type 1. Acta Biochimica et Biophysica Sinica, 2008, 40(1):. Chen N, Hu HX, Zhang ZF, Shuai JB, Fang WH. Genetic diversity of the envelope glycoprotein E2 of classical swine fever virus: Recent isolates branched away from historic and vaccine strains. Veterinary Microbiology, 2007, 127:. Shuai JB, Wei W, Li XL, Chen N, Zhang ZF, Chen XY, Fang WH. Genetic characterization of porcine circovirus type 2 (PCV2) from pigs in high-seroprevalence areas in southeastern China. Virus Genes, 2007, 35:. 陈健舜,江玲丽,方维焕。李斯特菌毒力因子及其进化探讨。微生物学报,2007, 47(4):738-742。14. 徐 程,陈亚波,方维焕。O型口蹄疫病毒结构蛋白VP1的原核表达与抗原性分析。动物医学进展,2007,28(6):14-18。15. 应薇芳,赵焕灿,钱 娅,方维焕。浙江部分地区屠宰场待宰猪链球菌的检测及药敏试验。动物医学进展,2007,28(6):24-26。16. 潘自降,坎 布,沈 飙,娄高明,方维焕。副溶血弧菌海产品和临床分离株的表型及溶血素相关基因分析。微生物学报,2007,47(3):508-511。17. 田国明,李肖梁,陈健舜,陈雪燕,方维焕。单核细胞增多性李斯特菌LMO1847基因在不同应激条件下的表达。畜牧与兽医,39(7):7-10。18. Chen XY, Xu JJ, Shuai JB, Chen JS, Zhang ZF, Fang WH. The S-layer proteins of Lactobacillus crispatus strain ZJ001 is responsible for competitive exclusion against Escherichia coli O157:H7 and Salmonella typhimurium. International Journal of Food Microbiology,2007,115(3):. Yang ZZ, Habib M, Shuai JB, Fang WH. Detection of PCV2 DNA by SYBR Green I-based quantitative PCR, Journal of Zhejiang University Science B, 2007, 8(3):. Jiang LL, Ke CL, Xu JJ, Chen JS, Chen XY, Chen N, Shuai JB, Fang WH. Listeria monocytogenes mutants carrying Newcastle disease virus F gene fused to its actA and plcB: In vitro expression and immunogenicity in chickens, Acta Biochimica et Biophysica Sinica, 2007, 39(1):57-6621. 陈亚波,徐 程,张桂芝,方维焕。新城疫病毒HN蛋白抗原表位分析及结构域基因原核表达。中国预防兽医学报,2007,29(1):32-3522. Li XL, Shuai JB, Fang WH*. Protection of Carassius auratus Gibelio against infection by Aeromonas hydrophila using specific immunoglobulins from hen egg yolk. Journal of Zhejiang University Science B, 2006, 7(11):. Yang ZZ, Fang WH, Habib M. First Results of Detection of PRRSV and CSFV RNA by SYBR Green I-based Quantitative PCR. Journal of Veterinary Medicine B, 2006;53(10):. Li L, Fang WH, Li JR, Huang YW, Yu L. Oral DNA vaccination with the polyprotein gene of infectious bursal disease virus (IBDV) delivered by attenuated Salmonella elicits protective immune responses in chickens. Vaccine, 2006, 24(33-34):. John Dikki M, Chen XY, Chen N, Fang WH*. Attenuated Salmonella typhimurium as a carrier for prokaryotic and eukaryotic expression vectors, Journal of Zhejiang University (Agric. & Life Sci., ed), 2006, 32(3):. Fang WH, Birgitte B, Siegumfeldt H. Leucocins 4010 from Leuconostoc carnosum cause a matrix related decrease in intracellular pH of Listeria monocytogenes, FEMS Microbiology Letters, 2006, 258(2): 208-21327. Jiang LL, Xu JJ, Chen N, Shuai JB, Fang WH. Virulence phenotyping and molecular characterization of a low-pathogenicity Listeria monocytogenes isolate from milk, Acta Biochimica et Biophysica Sinica, 2006, 38(4):. 徐晶靓, 江玲丽, 陈 宁, 帅江冰, 方维焕。携带外源基因的重组减毒单核细胞增多性李斯特菌构建与鉴定。微生物学报,2006,46(3):445-450。29. Vongxay KP, Cheng SY, Zhou XY, Shen B, He XL, Zhang GZ, Fang WH Prevalence of Vibrio parahaemolyticus in seafoods and their processing environments as detected by Duplex PCR. Journal of the Science of Food and Agriculture, 2006, 86(12): . 张桂芝,陈亚波,徐 程,张朝政,方维焕。新城疫病毒F蛋白结构域基因原核表达与抗原表位分析。浙江大学学报(农业与生命科学版)2006,32(1):82-87。31. 杨宗照, 方维焕。猪瘟合并猪繁殖呼吸综合征病毒(PRRSV)和猪圆环病毒2型(PCV2)感染。中国兽医学报, 2006,26(3):. Zeng HY, Zhang XF, Sun Z, Fang WH. Multiplex PCR identification of Listeria monocytogenes isolates from milk and milk-processing environments. Journal of the Science of Food and Agriculture, 2006, 86(3):. Yang ZG, Sun HX, Fang WH. Haemolytic activities and adjuvant effect of Astragalus membranaceus saponins (AMS) on the immune responses to ovalbumin in mice. Vaccine, 2005, 23 (44): . 李肖梁, 尹兆正, 钱娅,朱志刚,方维焕。抗嗜水气单胞菌IgY的提纯及体外抑菌效果研究。浙江大学学报(农业与生命科学版), 2005, 31(4):. 戴贤君, 方维焕。减毒沙门氏菌为载体的口服生长抑素DNA疫苗对草鱼安全性。中国兽医学报, 2005,25(5):. 戴贤君, 方维焕。减毒沙门氏菌为载体的草鱼口服生长抑素DNA疫苗构建及其稳定性。中国兽医学报 , 2005, 25(4):. 马有智,戴贤君,李肖梁,方维焕。表达猪链球菌溶血素基因的减毒沙门氏菌的构建及鉴定。中国兽医学报, 2005,25(5):478-480。38. 陈雪燕, John M. Dikki, 江玲丽, 帅江冰, 方维焕。重组质粒在减毒沙门氏菌中的稳定性及其对细菌侵袭力的影响。微生物学报,2005,45(5):. 贺晓龙, 张桂芝,方维焕。动物性食品中大肠埃希氏菌O157:H7特异性PCR方法的建立。中国兽医科技, 2005, 35(8):. Ke CL, Song HH , Jiang LL, Zeng HY, Zhang GZ, Fang WH. Construction of the recombinant Listeria monocytogenes mutant expressing green fluorescent protein. Journal of Zhejiang University (Agric. & Life Sci., ed), 2005, 31(5): . Jiang LL, Song HH, Chen XY, Ke CL, Xu JJ, Chen N, Fang WH. Characterization of a Mutant Listeria monocytogenes Strain Expressing Green Fluorescent Protein. Acta Biochimica et Biophysica Sinica, 2005, 37(1): 19–2442. Song HH, Wang ZL, Zheng DX, Fang WH, Li Y, Liu YY, Niu ZD, Qiu BS. A novel mucosal vaccine against foot-and-mouth disease virus induces protection in mice and swine. Biotechnology Letters, 2005, 27(21):. Song HH, Fang WH, Wang ZL et al. Detection of foot-and-mouth virus antibodies using a purified protein from the high-level expression of codon-optimized, foot-and-mouth disease virus complex epitopes in Escherichia coli. Biotechnology Letters, 2004, 26(16):1277-128144. 姜中其,陈晓红,方维焕,张应勤,孙建华。规模化猪场仔猪断奶腹泻大肠杆菌耐药性监测。浙江大学学报(农业与生命科学版),2004, 30(5):. 张晓峰,方维焕,程洁,施伟良。李斯特菌选择性增菌培养分离比较研究。中国公共卫生, 2004, 20(6):. 马有智,李肖梁,方维焕。猪链球菌2型溶血素基因PCR快速检测方法研究。浙江大学学报(农业与生命科学版)2004,30(1):78-8247. 李 龙,方维焕, 樊拥军,许 健,方 立,李建荣,于 涟。减毒沙门氏菌为载体在Vero细胞中表达传染性法氏囊病病毒多聚蛋白基因。生物工程学报,2004,20(3):437-440。48. Fang WH, Siegumfeldt H, Budde BB, Jakobsen M. Osmotic stresses led to decreased intracellular pH of Listeria monocytogenes as analyzed by fluorescence ratio-imaging microscopy. Applied & Environmental Microbiology, 2004, 70(5):. Song HH, Zhou L., Fang WH, Li Yong et al. High level expression of codon optimized food-and-mouth disease virus complex epitopes and cholera toxin B subunit chimera in Hansenula polymorpha. Biochemical and Biophysical Research Communications, 2004, 315(1):. Song HH, Li Y., Fang WH, Geng YF et al. Development of a set of expression vectors in Hansenula polymorpha. Biotechnology Letters, 2003, 25:. 谢荣辉,方维焕,李建荣,卢觅佳,于涟。萧山鸡IL-2基因真核表达载体构建及其在VERO细胞中的表达。浙江大学学报,2003,29(6):649-654。52. 梁雪芽,方维焕,江玲丽。鸡新城疫口服DNA疫苗的安全性、稳定性与免疫效力。 生物工程学报,2003,19(1):24-29。53. 马有智,方维焕,柯春林,张晓峰。猪链球菌2型JX分离株的生物学特性。中国兽医学报,2003,23(4):326-328。54. 马有智,方维焕。猪链球菌血清2型JX02株溶血素基因的克隆及序列分析。中国兽医学报, 2003,23(5):460-46255. 方维焕,梁雪芽,李建荣。减毒沙门氏菌为载体传递DNA疫苗诱导对鸡新城疫病毒的免疫保护。中国兽医学报,2003,23(6):527-529。56. 张晓峰,李爱云,方维焕,江玲丽,柯春林。多重聚合酶链反应鉴别单核细胞增生李斯特菌的研究。中华检验医学杂志,2003,26(2)93。57. 张晓峰,方维焕,江玲丽,杜华华。应用聚合酶链反应检测食品中单核细胞增多性李斯特菌。中国预防医学杂志, 2003,37(3):199。58. 方维焕。食品安全标准和监控体系建设需尽快与国际接轨。国际学术动态,2003,1:15-18。59. 张晓峰,程 洁,方维焕。抗李斯特菌免疫机理研究进展。中国人兽共患病杂志,2003,19(5):113-115。60. 方维焕、梁雪芽。减毒沙门氏菌为载体在真核细胞中表达鸡新城疫病毒F蛋白的研究。生物化学与生物物理学报,2002,34(4):488-493。61. Zhong BX, Weng HB, Fang WH. Preparation of protein samples for gel electrophoresis by sequential extraction. Journal of Zhejiang University, Science ed., 2002,3(5). 宋厚辉,方维焕。猪链球菌ZJ株溶血素基因的克隆和序列分析。中国预防兽医学报,2002,24(2):103-10563. 梁雪芽,方维焕。猪生殖与呼吸综合征病毒的分子生物学研究进展。中国兽医科技.2001,31(8).14-1764. 宋厚辉,方维焕。 产气荚膜梭菌主要毒力因子的分子生物学研究进展。预防兽医学进展,2001,3(1):13-16。65. Fang WH, Almeida ., & Oliver . Effects of lactoferrin and milk on adherence of Streptococcus uberis to bovine mammary epithelial cells. American Journal of Veterinary Research, 2000, 3:. Fang WH. & Oliver S. P. Identification of lactoferrin-binding proteins from bovine mastitis causing Streptococcus uberis. FEMS Microbiology Letters, 1999, 176:. Almeida ., Fang WH., & Oliver . Adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells are mediated by host cell proteoglycans. FEMS Microbiology Letters, 1999, 177:. Fang WH., Luther D. A. & Oliver S. P. Protein expression by Streptococcus uberis in co-culture with bovine mammary epithelial cells. FEMS Microbiol. Letters, 1998, 166:. Fang WH., Luther D. A., Almeida R. A. & Oliver S. P. Decreased growth of Streptococcus uberis in milk from mammary glands of cows challenged with the same mastitis pathogen. Journal of Veterinary Medicine B, 1998, 45:. Ali-Vehmas T., Vikerpuur M., Fang WH., and Sandholm M. Giving selenium supplements to dairy cows strengthens the inflammatory response to intramammary infection and induces a growth-suppressing effect on mastitis pathogens in whey. Journal of Veterinary Medicine A, 1997, 44:. Fang WH., Myllys V. & Sandholm M. Resazurin reduction as a function of respiratory burst of bovine polymorphonuclear neutrophils. American Journal of Veterinary Research, 1997, 58(6):. Fang WH. A novel fluorometric method for evaluation of postantibiotic effect on mastitis-causing Staphylococcus aureus and Escherichia coli. Journal of Microbiological Methods, 1996, 26:. Fang WH. Quantification of Staphylococcus aureus and Escherichia coli in the liquid medium by fluorometry and its use in phagocytosis assay. Journal of Appled Bacteriology, 1996, 80:. Koivunen A-L., Maisi P., Fang WH. & Sandholm M. Inhibition of the protease activity in tracheobronchial aspirates of horses with COPD. American Journal of Veterinary Research, 1996, 57(5):. Malbe M., Salonen M., Fang WH., Oopic T., Jalakas M., Klaassen M. & Sandholm M. Disposition of enrofloxacin (BaytrilR) into the udder after intravenous and intra-arterial injections. Journal of Veterinary Medicine A, 1996, 43:. Fang WH., Vikerpuur M. & Sandholm M. Reconstitution of mastitic milk by adding blood plasma and leukocytes into low cell count milk. Veterinary Research (France), 1996, 27:. Fang WH., Shi M., Huang L. & Chen J. Antagonism of the lactic acid bacteria towards Staphylococcus aureus and Escherichia coli on the agar plates and in milk. Veterinary Research (France), 1996, 27:. Fang WH. & Pyorala S. Mastitis-causing Escherichia coli: Serum sensitivity and susceptibility to selected antibacterials in milk. Journal of Dairy Science, 1996, 79:. Fang WH. & Vikerpuur M. Potency of antibacterial drugs in milk as analyzed by the ß-glucuronidase-based fluorometry. Journal of Veterinary Pharmacology & Therapeutics, 1995, 1:. Fang WH., Vikerpuur M. & Sandholm M. A fluorometric ß-glucuronidase assay for analysis of bacterial growth in milk. Veterinary Microbiology, 1995, 46:. Malbe M., Klaassen M., Fang WH., Myllys V., Vikerpuur M., Nyholm K., Sankari S., Suoranta K. & Sandholm M. Comparison of selenite and selenium yeast feed supplements on Se-incorporation, mastitis and leukocyte function in Se-deficient dairy cows. Journal of Veterinary Medicine A, 1995, 42:. Fang WH. & Sandholm M. Inhibition of the proteinase activity in mastitic milk. Journal of Dairy Research, 1995, 64:. Fang WH., Shi M., Chen J., Huang L. & Shao Q. Comparative aspects of bacterial growth in milk and whey. Chinese Veterinary Science (English), 1994, 1(1):. Fang WH., Shi M., Chen J., Huang L. & Shao Q. Turbidometric determination of bacterial growth in whey. Chinese Veterinary Science (English), 1994, 1(1):. Fang WH., Shi M., Huang L. & Shao Q. Growth of lactic acid bacteria, Staphylococcus aureus and Escherichia coli in normal and mastitic milk and whey. Veterinary Microbiology, 1993, 37:. Fang WH., Jiang C. & Liu H. Epidemiological aspects of bovine mastitis and its control in several dairy herds in south-east China. Preventive Veterinary Medicine, 1993, 15(2-3):. Hu SH, Fang WH, Lu HR, et al. Effect Of Teat Dipping And Dry Cow Therapy On Mastitis In A Commercial Dairy-Herd In China. Preventive Veterinary Medicine, 1990, 10:(1-2), 91-96.
1.先自我介绍,问各位评委好2.介绍题目,根据关键词介绍背景资料3.简明扼要讲技术路线图:本文设计以用水提取法和乙醇回流提取法,提取鹿衔草、牛耳大黄、扁蓄、岩陀的有效成分,通过体外抑菌试验筛选抑菌效果最好的中草药进行体内抑菌试验。4.应用步骤详细补充5.结合图表分析结果6.讨论及做出结论
1、特征,嗜水气单胞菌是一种广泛分布于自然界各种水的革兰氏阴性菌, ℃~℃为最适繁殖温度, 普遍存在于盐量较低的河水、海水、湖水、游泳池、下水道等环境中的条件致病菌,可感染多种水生动物和其他动物是人兽鱼共患的条件致病菌。2、鱼类,在水产中发现由嗜水气单胞菌感染的暴发性出血病较多,因嗜水气单胞菌的血清型很多,况且感染的对象不同,所以症状也各异。如白鲢暴发性出血病、甲鱼败血病、黄鳝出血病、鳗鲡红鳍病等。该菌为条件致病菌,当环境骤变,水质恶化时,常会与其它菌(如温和产气单胞菌、弧菌等)混合感染使病情加重。由嗜水气单胞菌感染的疾病一般病势较猛,多为恶性传染病,死亡率很高。3、大部分人感染嗜水气单胞菌一般只会导致急性胃肠炎,该病的发生与误食被污染的水源或食物而引起的肠道感染有关。少部分人由于皮肤的伤口接触到病菌后而发生外伤感染,轻者仅表现为局部的表浅溃疡,严重者感染可达皮肤深部组织,引起蜂窝织炎。免疫缺陷的人群,还可能引发严重的全身性感染性疾病,有较高的致死风险。
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