旨在利用杆状病毒表达系统构建1株对高致病性H7N9亚型禽流感病毒(A/Chicken/Huizhou/HZ-3/2016)攻击后的家禽提供保护的候选疫苗株.用同源重组的方法构建1株表达H7N9亚型禽流感病毒(A/chicken/Shaoxing/5201/2013株)血凝素(Hemagglutinin,HA)蛋白的重组杆状病毒.用PCR技术鉴定重组杆状病毒rBac-SX5201HA的遗传稳定性,用间接免疫荧光方法和Western Blotting方法鉴定HA蛋白的表达情况,用血凝试验检测HA蛋白的体外活性,继而对重组疫苗在无特定病原体(Specific pathogen free,SPF)鸡上进行免疫效力试验,并对免疫后21 d的SPF鸡血清进行抗体检测,对攻毒5 d后的鸡咽喉和泄殖腔棉拭子进行病毒分离.结果显示,重组杆状病毒rBac-SX5201HA在昆虫细胞中生长良好,且可稳定高效表达H7 HA蛋白,血凝效价可达26;重组疫苗免疫SPF鸡21 d后可诱导28.4血凝抑制(Hemagglutinin inhibition,HI)抗体效价并能抵抗高致病性H7N9亚型禽流感病毒的攻击,免疫组SPF鸡群均未发病或死亡,仅有17％的SPF鸡在攻毒后第5 d出现排毒.可以看出,重组疫苗对高致病性H7N9亚型禽流感病毒攻击后的SPF鸡提供了100％的保护,且可有效抑制病毒在SPF鸡体内的复制.

苯乳酸是一种能够存在于食品中的新型抑菌物质,为了避免传统高效液相色谱法(HPLC)检测苯乳酸时出现灵敏度低和色谱峰相连的问题,建立了一种二维高效液相色谱法(2D-HPLC)检测发酵食品中苯乳酸的方法,测定了酸奶、甜米酒、东北酸菜、酱黄瓜和广式腊肠5种常见发酵食品中苯乳酸的含量.利用超高效液相色谱-串联四级杆质谱(ultra high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)验证了样品中存在苯乳酸.2D-HPLC方法:第一维色谱柱为C18色谱柱,流动相为0.1％甲酸和甲醇;第二维色谱柱为C8色谱柱,流动相为0.05％三氟乙酸和乙腈,流速1.0 mL/min,柱温30℃,检测波长210 nm.结果 表明:该方法在0.4～10.0 mg/L内呈现良好的线性关系,r=0.999 9,检出限0.16 mg/L,定量限0.52 mg/L,方法的日内和日间重复性相对标准偏差(relative standard deviation,RSD)分别为0.60％和0.94％,5种发酵食品的加标回收率91.79％ ～ 107.37％,RSD ＜7.5％;UPLC-MS/MS验证样品中含有苯乳酸.利用2D-HPLC法测定食品中的苯乳酸前处理简单、结果准确、抗干扰能力强,适用于大批量样品检测,为2D-HPLC法检测发酵食品中组分提供理论依据.

目的 探讨辣木叶总黄酮提取物抗疲劳效果及其生化机制.方法 将48只雄性昆明小鼠分为正常对照(NC)、辣木叶粗提物(MLCE)和辣木叶总黄酮提取物(MLFE)3组.NC组灌胃纯水,MLCE和MLFE组分别灌胃辣木叶粗提物溶液[340 mg/ (kg· bw)]和辣木叶总黄酮提取物溶液[75 mg/ (kg·bw)],体积均为0.5ml,灌胃14d后负重游泳,记录力竭游泳时间;另将48只小鼠分为安静对照(QC)、游泳对照(SC)和辣木叶总黄酮提取物(MLFE)3组,QC和SC组灌胃纯水,MLFE组灌胃辣木叶总黄酮提取物溶液[75 mg/ (kg·bw)],灌胃体积均为0.5ml.实验D15,SC和MLFE组非负重游泳,90 min后取血液、肝脏和后腿肌肉组织,QC组采血后取相关组织,测定生化指标.结果 MLFE显著延长力竭游泳时间,且升高了游泳后小鼠血糖(Glu),降低血乳酸(LA)、尿素氮(BUN)、丙二醛(MDA)水平,提高血清过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性;维持肌肝糖原水平,提高组织中抗氧化酶、ATP酶活性及肌肉中游离脂肪酸(NEFA)的含量.结论 辣木叶总黄酮提取物能通过调节能量代谢,减少代谢产物积累、提高抗氧化能力、动员脂肪等途径来提高运动耐力,延缓疲劳发生.

为建立一种反相高效液相色谱法(Reversed phase high performance liquid chromatography,RP-HPLC)检测乳酸菌发酵液中苯乳酸含量的方法,系统考察了3种固定相、2种流动相有机相、5种水相及其比例对苯乳酸RP-HPLC检测的影响并对方法进行了评价.确定了以双封端C18色谱柱(4.6 mm × 250 mm,5μm)为固定相,0.05％三氟乙酸(Trifluoroacetic acid,TFA)-乙腈,体积比75∶25,等度洗脱,流速1 mL/min,检测波长210 nm,柱温30℃.在0.2～5.0 mg/L内线性良好,r=0.9998,回收率99.88％ ～101.08％,检出限0.1400 mg/L,定量限0.4600 mg/L,ld精密度和重复性相对标准偏差(Relative standard deviation,RSD)为0.30％和0.64％,30 d精密度和重复性RSD为0.59％和0.79％,30 d内苯乳酸标准储备液稳定性RSD为2.45％,乳酸菌发酵液中苯乳酸合成量为0.1400 mg/L时,其扩展不确定度为0.0046,95％的置信区间,包含因子k=2.选用短乳杆菌P3(Lactobacillus brevis)和植物乳杆菌(Lactobacillus plantarum)ATCC8014发酵液为检测对象,培养120 h苯乳酸合成量分别为77.58和62.60 mg/L.结果 表明,在该方法下苯乳酸能够稳定出峰,检测效率高,检测成本低,操作简单,结果准确可靠,适合检测大批量样品,可为苯乳酸的相关研究提供理论依据.

地方高水平特色大学如何按照“双一流”要求发展,实现自身的建设目标,需要学校统筹谋划,将自身置于世界的坐标中自我审视、自我定位、自我革新、自我提高,以世界一流大学和一流学科的标准为指引,借鉴世界一流大学的成功经验,并结合学校自身情况制定相应的举措,推进学校的建设,实现自我彰显、自我提高.研究在比较分析的基础上,以上海海洋大学为例,结合学校发展的实际,简述学校在“双一流”建设中的战略设计与发展策略.

类黄酮是植物界中分布最广泛的天然化合物之一,具有多种功能活性,如抗氧化、抗菌、抗炎等,还可以调节血糖、血脂和血压,因此广受人们的关注与研究.本文对植物类黄酮的提取纯化方法进行回顾,概述了溶剂提取法、酶辅助提取法、超声波辅助提取法、微波辅助提取法、超临界CO2萃取法等提取方法以及有机溶剂萃取法、大孔吸附树脂吸附法、聚酰胺树脂吸附法、制备型高效液相色谱法和高速逆流色谱法等纯化方法,旨在为今后植物类黄酮的研究和开发提供参考.

研究了Lactobacillus brevis对水溶液中Cr(Ⅲ)的吸附作用.考察了初始pH值、接触时间、初始Cr(Ⅲ)浓度、菌体浓度和温度对Cr(Ⅲ)吸附效果的影响.结果表明,在较低的pH、温度和初始Cr(Ⅲ)离子浓度条件下,菌株对Cr(Ⅲ)离子的吸附量较低.在试验条件下,溶液初始pH、温度和初始Cr(Ⅲ)离子浓度的升高,均能提高菌株对Cr(Ⅲ)离子的吸附量.在温度为40℃、pH 6和初始Cr(Ⅲ)离子浓度为200 mg·L-1时,菌株吸附量最大.随菌体浓度升高,单位浓度菌体对Cr(Ⅲ)离子吸附量降低,但总吸附量增大,菌体浓度为6 g·L-1吸附量最大.菌株对Cr(Ⅲ)离子吸附较快,接触时间为1h就达到平衡.用Langmuir、Freundlich、Redlich-Peterson和Temkin吸附模型进行拟合,相关的回归系数表明,吸附过程拟合Langmuir吸附模型比Freundlich、Redlich-Peterson和Temkin吸附模型好.用Elovich、准一级、准二级动力学拟合,动力学试验数据与Lagergren准二级动力学方程的拟合度最佳.

地方高水平特色大学如何按照"双一流"要求发展,实现建设目标,需要学校统筹谋划,将自身置身于世界的坐标中自我审视、自我定位、自我革新、自我提高,以世界一流大学和一流学科的标准为指引,借鉴世界一流大学的成功经验,并结合学校情况制定相应的举措,推进学校的建设,实现自我彰显、自我提高.本文在比较分析的基础上以上海海洋大学为例,结合学校发展的实际,简述学校在"双一流"建设中的战略设计与发展策略.

In order to optimize degradation conditions of fermented grains on tetrodotoxin(TTX),24-days fermented grains were used as degradable material, the influence of degradation conditions was investigated, such as dosage of fermented grains, degradation temperature, degradation time, and shaking speed. The mouse bioassay was used to evaluate the degradation effect of the fermented grains under the optimal conditions.It was found that when the fermented grains was 16 g, the temperature was 30℃,and the rotate speed was 50 r/min,the fermented grains had the best degradation ability;the death time of Kunming mice was prolonged with the extension of TTX degradation time under the optimal conditions,and then remained unchanged after five days.

Lead-resistant and adsorptivity lactic acid bacteria were screened out from pickles and used for the removal of heavy metal lead.Four strains were isolated from the samples by spread plate method,and lead resistant reached 2000 mg/L,when the concentration of lead was 100 mg/L,,the adsorption rate of p3 was highest,reached 78.90％.The results of morphology observation,physiological and biochemical identification and mass spectrometry detection and 16S ribosomal deoxyribonucleic acid sequencing data showed that p3 could be identified as Lactobacillus brevis.The strain grew well in the salt containing 0～5％,and it was able to tolerate the hydrochloric acid with pH value of 2,the salts of 8.0 g/L and trypsin of 1.1 g/L.It can be used to treat waste water,make pickles orreduce lead levels in animals as a probiotic in the future.

目的:对于黄酒酒曲样品,选择一种合适的总DNA提取方法,并对其中真菌群落结构进行多样性分析.方法:因其质地,结合以往DNA提取方法,比较分析不同方法所获得的总DNA纯度、得率及凝胶电泳图,对实验结果较好的DNA样本进行真菌引物PCR扩增后纯化定量,再经Miseq高通量测序后进行生物信息分析.结果:实验结果表明,使用试剂盒+物理化学酶法获得了较高质量的样品总DNA,A260/A280比值也接近理想水平,PCR扩增后凝胶电泳图能够得到目的条带,OTU分类、Shannon曲线和系统进化树图显示了其真菌多样性.结论:通过数据的比较和分析,确立了选择的方法是一种较理想的酒曲总DNA的提取方法,对其真菌群落的测定及分析,也为后期开拓黄酒微生物资源提供了有力工具.

国内外对金黄色葡萄球菌预测模型的研究集中处于一、二级模型,且乳品方面的研究又集中在牛奶,而微生物在搅打奶油的生长模型研究基本处于空白.为了填补金黄色葡萄球菌在搅打奶油中生长行为变化的空白,为金黄色葡萄球菌的风险评估提供数据支持.本文选用产A型肠毒素的金黄色葡萄球菌作为研究对象,测定了15℃、20℃、25℃、30℃、36℃下金黄色葡萄球菌在新鲜奶油中的生长曲线,运用Origin8.0软件和DMFit软件,分别使用修正的Gompertz、修正的Logistic和Baranyi三模型来拟合不同温度下的生长模型.通过比较三种模型的拟合结果,选择最适模型的生长动力学参数,选用平方根方程构建金黄色葡萄球菌最大比生长速率与温度的二级模型.结果显示,Baranyi模型对金黄色葡萄球菌的生长曲线拟合效果最好,构建的二级平方根模型拟合度较好:R2为0.92,准确因子(Af)和偏差因子(Bf)均在1左右.所构建的二级模型,能真实快速有效地预测15～36℃金黄色葡萄球菌的生长情况.

随着高等教育的分化,地方特色型高校成为我国高等教育的重要办学力量.在转型发展中,建设高水平特色大学成为这些高校的建设目标.加强优势特色学科建设是提升学校核心竞争力的关键,通过优势学科带动相关、支撑学科的发展.通过分析地方特色型高校开展学科国际评估的需求,结合上海海洋大学学科建设与评估的思路,思考地方特色型高校学科国际评估指标体系构建的要点.

[目的]建立一种快速、稳定、可靠的海洋病毒计数方法.[方法]海水水样经福尔马林固定后,滤过孔径为0.02 μm的Anodisc Al2O3膜.滤膜经SYBR Green Ⅰ染色后,在相应波长的激发光下进行观察.借助荧光显微镜目镜网格尺,计数视野中的病毒颗粒,换算后获得样品中病毒的浓度.[结果]对具体实验方法进行了优化,可快速、稳定地对海水中的病毒计数.[结论]建立了一种适用于国内实验条件的、可靠的海洋病毒计数方法.

Metagenomic technology is applied in studying the total genomes of complex microbial community in specific environment.It overcomes the limitations of conventional methods of microorganism cultivation,and provides powerful techniques to explore and utilize rich gene resources of unculturable microorganisms.Remarkable progress has been achieved in this research field.This paper reviewed the theory and application of metagenomic technology in screening novel functional enzymes from microorganisms.

为研究污水处理活性污泥微生物多样性,提取了活性污泥宏基因组DNA,并采用细菌通用引物27F和1492R扩增了上海污泥厂活性污泥细菌16S rDNA片段,构建了细菌16S rDNA克隆文库,并对该文库中的微生物群落进行了分析.共获得200条高质量序列并建立系统发育树,结果显示活性污泥主要的细菌类群为变形菌门(Proteobacteria)(91.9％)、厚壁菌门(Firmicures)(4.6％)、拟杆菌门(Bacteroidetes)(2％)、绿弯菌门(Chloroflexi)(0.5％)、硝化螺菌门(Nitrospirae)(1％).其中,明显的优势菌群为Alcaligenes feacalis(55％)、Pseudomonas aeruginosa(12.8％)和Stenotrophomonas(12.8％),优势菌的产酶能力在活性污泥中显示生态修复功能菌的作用.

The aim of this study was to investigate virulence and drug resistant mechanism of multidrug resistant Pseudomonas aeruginosa(PA) isolated from water sample of fishery pond,in order to provide scientific support for practical aquaculture disease treatment,effective management of aquaculture environment and aquatic food safety control.Antimicrobial susceptibility of PA isolates under this study were examined by using standard Kirby Bauer disk diffusion method according to the Clinical and Laboratory Standards Institute(USA,2010 Edition),showing distinct susceptibility to nine antimicrobial agents belonging to six drug classes tested.All PA isolates were resistant to trimethoprim and rifampin,of which,90% were to ampicilin and kanamycin,and 10% were to gentamycin.The genes involved in pathogenicity,intrinsic and transferred antibiotic resistance were detected by polymerase chain reaction(PCR).The toxA and exoU genes were absent from the PA isolates tested,50% of which were exoS+/exoU-infectious genetype,and none of which were exoS-/exoU+ cell toxic genetype that usually existed in clinical PA isolates.Subsequent analysis revealed that the multidrug efflux pumps including MexAB-OprM,MexXY-OprM and MexVW-OprM were present in all PA isolates,20% of which were positive for ampC gene,whereas the other four efflux pumps including MexEF-OprN,MexJK-OprM,MexCD-OprJ and MexGHI-OpmD were fully or partially deficient.In addition,the comINT gene of class Ⅰ-Ⅲ integrons was negative in the PA isolates,however,the functional genes(int,soj,pilS2,pilD)of conserved module structures of integrative conjugative elements(ICEs) were detected positive,suggesting possible transfer activity of the ICEs detected in the PA isolates.This study revealed the major mechanism under multidrug resistance of the PA isolates mediated by efflux pumps encoded by chromosome genes,and constituted the first evidence for mobile genetic element ICEs in PA isolates of environmental origin.

In summer of 2008,green tides broke out in south Yellow Sea.First,the nutrition of floating algae was analyzed and then the algae were treated by sulphuric acid for getting reducing sugar.The effects of temperature(℃),treating time(min),sulphuric acid concentration(%),ratio of mass weight(%)on the biomass hydrolyzation were studied and the hydrolysis conditions were optimized by orthogonal analysis.The results showed that optimum conditions for floating Ulva prolifera biomass hydrolysis by sulphuric acid were 90 ℃ of treating temperature,70 min of treating time,5% of sulphuric acid concentration,4.5% of ratio of the mass weight.Liquid chromatography analysis showed that the molar ratio of monosaccharides in hydrolyzate were glucose 1.71∶ xylose 1.00∶ rhamnose 1.29.Growth curves of five yeasts(IwSc1,S1,S2,1770,1766) were drawn in order to ferment the liquid at stationary phase.The results showed that S2 was the best for getting highest ethanol concentration with GC analysis.The ethanol concentration was 2.1 g/L and the ethanol conversion rate was 26%.Glucose concentration dropped from 18.26 g/L to 2.38 g/L and the utilization rate of it was 87%.S1,S2 and IwSc1 mainly use glucose compared with 1770,1766 which can use both glucose and xylose.

A confirmative method was developed with high performance liquid chromatography-mass spectrometry(HPLC-MS/MS) to detect acyl-homoserine lactoned (AHLs) secreted by bacterial quorum sensing. The AHLs secreted by bacterial quorum sensing were extracted with ethyl acetate. The samples could be directly analyzed after a simple treatment. Sunfire C(18)column(2. 1 mm x 50 mm, 3.5 mu m) with methanol and water (including 2 mmol/L ammonium acetate, 0. 1% formic acid) as the mobile phase was used for separation. The detection method of electrospray ionization source with positive ion was used for mass spectrometric analysis. Qualitative and quantitative analysis of 11 kinds of AHLs were completed within 12 min. Experimental results showed that, the AHLs was in good linearity(r > 0. 996) in the range of 1 - 250 mu g/L. The detection limit of the method was 0. 1 - 1. 0 mu g/kg. The limits of quantification were 0. 3 - 3. 0 mu g/kg. The average recoveries were 54.4% - 128. 6%. The relative standard deviations were 3. 5% - 13. 9%. The results show that, this method is simple, rapid, accurate, and highly sensitive to analyse and detect the AHLs in the bacterial growth environment.

Irradiation is a physical method that uses exposure energy of atomic energy beam to sterilize and preserve food.Due to its characteristics of environmental protection,safety and high efficiency,this method that can efficiently restrain or eliminate pathological or spoiled organisms has been widely applied in sterilization and preservation of meat products.In this paper,mechanisms of radiation sterilization,current research progress of food irradiation at home and abroad,safety evaluation of food irradiation and irradiation standards for meat products are reviewed.Meanwhile,the research progress of 60Co γ-ray irradiation,electron beam irradiation and X-ray irradiation in sterilization and preservation of meat products are discussed.Moreover,effects of irradiation on sterilization are also summarized.Furthermore,the feasibility and advantages of applying X-ray irradiation in sterilization and preservation of meat products are also explored.Therefore,a future possibility for applying X-ray irradiation as a pilot technology in the field of food is proposed.