人工模拟肠炎沙门氏菌在绿豆芽中的内化定殖能力及其消毒处理研究

本研究以绿豆为作用载体,研究了肠炎沙门氏菌(ATCC13076)在绿豆芽的不同发芽阶段中的内化定殖能力及消除绿豆芽中定殖的肠炎沙门氏菌的有效方法。结果表明:在绿豆发芽前期的4个不同时段(0~48 h内):吸胀期(G_1,0 h)、萌动期(G_2,12h)、发芽初期(G_3,24 h)、发芽期(G_4,48 h),分别接种10~2、10~4、10~6和10~8 CFU/mL的肠炎沙门氏菌时,其在绿豆芽中的内化定殖能力不同。接种浓度为10~2CFU/mL左右时,肠炎沙门氏菌在发芽初期(G_3)和发芽期(G_4)的内化能力较强,另外两个阶段比较微弱。接种浓度不低于10~4CFU/mL时,肠炎沙门氏菌在吸胀期(G_1)的内化能力最强。在吸胀期(G_1)接种10~8CFU/mL的菌液时,最高内化量可达2.6×10~8CFU/g。紫外照射消毒处理对内化定殖的肠炎沙门氏菌有明显的去除效果,而次氯酸钠溶液和硝酸银溶液浸泡处理对内化的肠炎沙门氏菌的去除效果并不明显。     

GB 4789.28食品微生物学检验用培养基和试剂中沙门氏菌检验用培养基质控菌株的筛选

该研究根据《GB 4789.28-2013 食品安全国家标准 食品微生物学检验 培养基和试剂的质量控制》（以下简称GB 4789.28-2013）的检测要求,从我国菌种库中筛选出1株鼠伤寒沙门氏菌可等效于GB 4789.28-2013中的标准菌株（ATCC14028）,用于评价培养基的质量。该研究联合15家验证机构用7株不同分离源的鼠伤寒沙门氏菌,依据GB 4789.28-2013的检验方法验证不同品牌的培养基,将生长率、选择性与ATCC41028一致性最高的菌株选为标准的等效质控菌株。为确保结果的可重复性和一致性,各实验均采用同品牌同批次的培养基进行验证,每株菌株均由至少5家单位进行验证。综合生长率与选择性的结果,ATCC41028在各品牌培养基上均可良好生长,CMCC(B)50976与ATCC41028结果的一致性最高,而CMCC(B)50976最难进行培养或分离。同时,结果显示,不同品牌、不同人员、不同实验室环境下进行的培养基验证结果均有差异。工作组从7株鼠伤寒沙门氏菌中筛选出CMCC(B)50976作为GB 4789.28的质控菌株,可等效于国外的标准菌株（ATCC14028）,新菌株更具有代表性,且减少了国外菌株运输过程中的生物风险。     

雾化植物精油对鼠伤寒沙门氏菌抑制效果和樱桃番茄品质的影响

本研究的目的是评测雾化液态植物(芥末、肉桂、牛至和百里香)精油对樱桃番茄表面和茎端的抑菌效果及对果实品质的影响。将樱桃番茄的表面和茎端分别接种鼠伤寒沙门氏菌(ATCC 53467和ATCC 5 3468菌种),晾干之后放到包装盒中,采用加湿器来雾化液态的植物精油,然后通过一根PE管导入到装有已经接种了鼠伤寒沙门氏菌的樱桃番茄的包装盒中。处理后的果实存放在10℃的冰箱中3个星期,存放期间每隔1个星期进行微生物指标测定,不接菌种的果实用来测定果实品质指标。结果表明,雾化的植物精油能够显著的抑制樱桃番茄表面和茎端的鼠伤寒沙门氏菌(表面及茎端菌落分别降低2.00 log CFU/fruit和1.00 log CFU/fruit左右)。随着贮藏期的延长,菌落总数也随之降低,这证明其抑菌效果仍然有效。雾化植物精油杀菌处理对果实硬度,颜色,VC等品质指标没有显著影响(p0.05)。雾化是一种方便有效的可用于储存或运输果蔬产品消毒的新方法。     

香瓜片鼠伤寒沙门氏菌生长控制

为了有效控制香瓜片等生食水果携带鼠伤寒沙门氏菌的生长,采用物理(超高压)、生物(蛭弧菌BDM01)2种方法对香瓜片上接种的鼠伤寒沙门氏菌进行消除控制并比较杀菌效果及处理后的香瓜片感官状态。结果表明,与对照组相比,2种方法对香瓜片鼠伤寒沙门氏菌均有很好的控制作用(p<0.05);蛭弧菌控制鼠伤寒沙门氏菌的效果可维持24 h,且在9 h(宿主菌:BD=1:1)和12h(宿主菌:BD=1:10)出现明显的致病菌数目的下降,同时其感官状态在12 h内一直保持良好。超高压杀菌效果在最初非常明显,但在随后的储存实验中,致病菌数目迅速增长,且香瓜片在经过超高压作用后,其感官状态逐步下降。结果揭示,蛭弧菌BDM01可有效控制鲜切香瓜片鼠伤寒沙门氏菌的生长。     

富硒绿豆芽饮料研制

将绿豆用50~75mg／kgSe溶液在35℃下浸泡6h,然后经5d发芽生长制成绿豆芽;再将此绿豆芽制成了风味独特的富硒豆芽饮料。     

草鱼鱼糜中沙门氏菌生长预测模型的建立

为建立鱼糜中沙门氏菌生长预测模型,选用新鲜草鱼鱼糜和鼠伤寒沙门氏菌作为研究对象,比较了4、10、20、28、37℃条件下鱼糜中鼠伤寒沙门氏菌的生长情况,分别采用Huang模型,Baranyi模型和修正的Gompertz模型进行拟合,建立鱼糜中鼠伤寒沙门氏菌一级生长动力学模型。并用平方根模型方程描述温度与比生长速率和延滞期的关系,得到鼠伤寒沙门氏菌生长二级模型。使用判定系数R2,准确因子(Af),偏差因子(Bf)和均方误差(MSE)对一级和二级模型可靠性进行评价,结果表明修正的Gompertz模型更适合于描述4~37℃条件下鱼糜中鼠伤寒沙门氏菌的生长变化,二级平方根模型可用于描述鱼糜中鼠伤寒沙门氏菌的生长参数,能够为鱼糜中沙门氏菌的监测提供一定的参考依据。     

乙醇适应对鼠伤寒沙门氏菌及其菌膜耐致死胁迫的影响

本文主要研究鼠伤寒沙门氏菌在玻璃表面生物菌膜(BF)的生长特性和乙醇作用方式对鼠伤寒沙门氏菌菌膜形成的影响以及乙醇适应处理对浮游菌与菌膜耐致死胁迫的作用。采用结晶紫染色观察鼠伤寒沙门氏菌菌膜的形成,用酶标仪测量波长570 nm处不同条件下生物菌膜的生物量,用平板菌落计数衡量乙醇适应处理对浮游菌及菌膜耐致死胁迫的影响。结果表明,结晶紫染色可清晰观察到鼠伤寒沙门氏菌在玻璃表面形成的生物菌膜,菌膜形成的紧密网状结构随着时间的延长而增强。乙醇作用方式对菌膜形成有显著影响,加入不同含量的乙醇后培养48 h,乙醇对菌膜形成有显著的抑制作用。预先培养菌24 h后加入5%的乙醇,能显著促进菌膜的生长。菌膜形成过程中,当营养条件为1/10 TSB,5%乙醇适应能增加菌体对苹果酸的耐受性,增加浮游菌对12%的乙醇及5 mg/mL苹果酸的耐受性。     

采用紫外线照射控制发芽糙米细菌总数研究

发芽糙米在发芽过程中微生物大量生长,存在安全隐患;本研究采用紫外线照射方法以控制发芽糙米细菌数量。结果表明,紫外光可有效减少发芽糙米中微生物数量,糙米在发芽24 h后,每隔6 h采用距紫外灯(30 W)光源25 cm距离照射3 min处理,发芽糙米细菌总数保持10~5cfu/g以下,发芽率没大幅下降,芽长增加,γ-氨基丁酸含量增加,且发芽糙米感官质量有明显提高。     

实时荧光PCR法检测鼠伤寒沙门氏菌

目的 建立实时荧光PCR法检测鼠伤寒沙门氏菌的方法。方法 基于鼠伤寒沙门氏菌II型限制酶基因, 设计引物及Taqman探针, 利用实时荧光PCR进行特异性、灵敏性及模拟样品的检测实验。结果 特异性探针可从25种血清型沙门氏菌(共49株)及11株阴性对照菌株中检测出全部的11株鼠伤寒沙门氏菌。以鼠伤寒沙门氏菌梯度稀释菌液DNA为模板进行实时荧光PCR实验, 菌株模板浓度与Ct值呈良好线性关系, 线性系数(R2)为0.998, 扩增效率90%, 最低检测浓度300 cfu/mL。对已接种鼠伤寒沙门氏菌的4种模拟样品同时进行实时荧光PCR检测和传统方法鉴定, 两者结果一致。结论 此方法特异、灵敏、准确, 适于食品中鼠伤寒沙门氏菌的检测。     

一株强抑菌活性植物乳杆菌的分离及益生性能研究

以健康黔北黑猪肠道黏膜内容物为原料,采用厌氧罐培养法分离出肠道细菌,结合抑菌试验筛选出其中具有强抑菌活性菌种进行分子生物学鉴定,并对其耐受性及抑菌物质的稳定性进行研究。结果表明,该菌株被鉴定为植物乳杆菌（Lactobacillus plantarum）,对金黄色葡萄球菌ATCC6538、大肠埃希氏菌ATCC25922和鼠伤寒沙门氏菌ATCC14028等指示菌具有强抑制作用,其发酵上清液对3种指示菌的最大抑菌圈直径分别约为44.7 mm、23.1 mm和31.8 mm;菌株发酵上清液抑菌物质耐高温,120 ℃处理30 min抑菌活性无显著变化（P＞0.05）;胰蛋白酶处理后,上清液的抑菌圈直径显著减小（P＜0.05）,而过氧化氢酶和胃蛋白酶处理后抑菌圈直径无显著变化（P＞0.05）,表明主要抑菌物质为有机酸和活性肽;菌株经pH 2、0.3%猪胆盐或人工胃肠液处理6 h后存活率分别为0.06%、0.61%、40.80%和25.65%,显示出良好的益生性能。     

绿豆萌发过程中绿豆蛋白的功能特性及其抗氧化性

本文以绿豆为材料,研究了其萌发过程中绿豆蛋白的功能特性(溶解性、持水性、持油性、乳化性、起泡性、乳化稳定性、起泡稳定性)及抗氧化性(DPPH自由基清除率、金属离子螯合率、超氧阴离子自由基清除率)的动态变化。结果表明,随着萌发时间的不断延长,绿豆蛋白的溶解性、持水性、持油性、乳化性、起泡性均呈现先增加后降低的趋势,乳化稳定性和起泡稳定性得以增强。其中,溶解性萌发24 h时达到最高,萌发96 h最低;萌发的绿豆蛋白持水性、持油性和乳化性相对于未萌发的分别提高了1.57倍、4.13倍和2.47倍;乳化稳定性、起泡性和起泡稳定性较未萌发的分别提高了43.8%、46.6%和61.3%。此外,萌发过程中的绿豆蛋白抗氧化性呈现先升高后下降的趋势,萌发促进了绿豆蛋白的抗氧化性。其中,DPPH自由基清除率和金属离子螯合率均在绿豆萌发36 h达到最大,较未萌发的分别提高了73.8%和31.0%;超氧阴离子自由基清除率萌发48 h达到最大,较未萌发的提高了81.7%。随着绿豆萌发时间的延长,绿豆蛋白的功能特性和抗氧化性呈现先升高后下降的趋势,萌发中期(24~48 h)达到最大。因此,萌发提升了绿豆蛋白的功能特性和抗氧化性,扩大了其在食品加工中的应用,提高了其利用价值。     

Effect of Soaking and Germination on Polyphenol Content and Polyphenol Oxidase Activity of Mung Bean (Phaseolus Aureus L.) Cultivars Differing in Seed Color

Five mung bean cultivars varying in seed coat color were analyzed for polyphenol content. The effect of soaking and germination on the polyphenol content and polyphenol oxidase activity was investigated. The level of total polyphenols was found to be in the range of 310–340 mg/100 g in whole seeds. Soaking reduced the total polyphenols, whereas germination for 48 h increased total polyphenol content by 41–76%. The hydrolysable tannins, condensed tannins, and hydrolysable tannins/condensed tannins index increased with an increase in period of germination. The total and percent condensed tannins content slightly decreased while hydrolysable tannins increased during germination. Maximum polyphenol oxidase activity of 102–108 units was observed in 24 h germinated seeds. There was no significant correlation found between the polyphenolic content and polyphenol oxidase activity in germinating mung bean seeds. These findings demonstrated that the mung beans are fair sources of polyphenols, thus having great potential as a source of natural antioxidants.     

Spatial distribution of Salmonella, Escherichia coli O157:H7, and other bacterial populations in commercial and laboratory-scale sprouting mung bean beds

The reliability of testing spent irrigation water to assess the microbiological status of sprouting mung bean beds has been investigated. In commercial trials, the distribution of opportunistic contaminants within 32 bean sprout beds (25 kg of mung beans per bin) was assessed 48 h after germination. The prevalence of generic Escherichia coli, thermotolerant coliforms, and Aeromonas in sprouts (n = 288) was 5, 11, and 39%, respectively, and 57, 70, and 79% in the corresponding spent irrigation water samples (n = 96). Contamination was heterogeneously distributed within the seedbed. In laboratory trials, beans inoculated with a five-strain cocktail of either Salmonella or E. coli O157:H7 (10(3) to 10(4) CFU/g) were introduced (1 g/500 g of noninoculated seeds) at defined locations (top, middle, or base), and the beans were then sprouted for 48 h. When seeds inoculated with pathogens were introduced at the base or top of the seedbed, the pathogens were typically restricted to these sites and resulted in 44% of the spent irrigation water samples returning false-negative results. Introducing inoculated beans into the middle or at the presoak stage enhanced the distribution of both pathogens within the subsequent sprout bed and resulted in comparable levels recovered in spent irrigation water. The study demonstrated that even though screening a single sample of spent irrigation water is more reliable than testing sprouts directly, it does not provide an accurate assessment of the microbiological status of sprouting mung bean beds. Such limitations may be addressed by ensuring that bean batches are mixed prior to use and by taking spent irrigation water samples from multiple sites at the latter stages of the sprouting process.     

Interaction of Salmonella enterica subspecies enterica serovar Typhimurium and mung bean (Phaseolus aureus) plants

The effect of Salmonella enterica subspecies enterica serovar Typhimurium, a zoonotic serovar, on mung bean (Phaseolus aureus) cultivar Pant Mung-3 plants was studied. Inoculation of mung bean seeds with Salmonella Typhimurium (7.2 x 10(5) CFU/ml) reduced sprouting rate (P < 0.07). This effect was more pronounced at higher levels of contamination. In the soil inoculated with Salmonella Typhimurium (7.2 x 10(6) CFU/g), germination was retarded and the number of defective sprouts was also significantly higher (P < 0.002). Salmonella Typhimurium grew inside germinating seeds and plant tissues and persisted in seedlings, adult plants, and harvested seedlings dried and stored at room temperature (30 degrees C) up to 45 days. Phaseolus aureus plants grown in sterile soil was resistant to Salmonella Typhimurium infection at 15 days of age and cleared Salmonella from all the aerial parts within 3 h of infection. However, Salmonella Typhimurium could be reisolated from the basal area of the stem and from soil even after 45 days of exposure to the pathogen.     

Reduction of Escherichia coli O157:H7 and Salmonella spp. on laboratory-inoculated mung bean seed by chlorine treatmentt

Three U.S. outbreaks of foodborne illness due to consumption of contaminated raw mung bean sprouts occurred in the past 2 years and were caused by Salmonella Enteritidis. The original source of the pathogens is thought to have been the seed. The aim of this study was to determine whether treatment with aqueous chlorine would eliminate the pathogens from mung bean seed inoculated in the laboratory with four-strain cocktails of Escherichia coli O157:H7 and Salmonella spp. Treatments (for 5, 10, or 15 min) with buffered (500 mM potassium phosphate, pH 6.8) or unbuffered solutions containing 0.3 or 3.0% (wt/vol) Ca(OCl)2 were tested. In order to mimic common commercial practice, seed was rinsed before and after treatment with sterile tap water. Treatment for 15 min with buffer (500 mM potassium phosphate, pH 6.8) or sterile water in combination with the seed rinses resulted in maximum reductions of approximately 3 log10 CFU/g. The largest reductions (4 to 5 log10 CFU/g) for the chlorine treatments in combination with the rinses were obtained after treatment with buffered 3.0% (wt/vol) Ca(OCl)2 for 15 min. Treatment of mung bean seed for 15 min with unbuffered or buffered 3.0% (wt/vol) Ca(OCl)2 did not adversely affect germination. Even though treatments with 3% (wt/vol) Ca(OCl)2 in combination with the water rinses were effective in greatly reducing the populations of both bacterial pathogens, these treatments did not result in the elimination of the pathogens from laboratory-inoculated seed.     

萌发绿豆中多酚类物质动态变化规律及其抗氧化活性的研究

以产自山西的明绿豆为实验材料,探究萌发前后绿豆中酚类化合物的动态变化规律及其抗氧化活性。采用传统方法进行绿豆的萌发,通过超声-微波协同萃取法提取绿豆及绿豆芽中的多酚类化合物,结合植物广靶代谢组学的方法对绿豆萌发前后多酚类化合物的提取液进行定性定量分析,明确酚类化合物的组成,分析萌发处理对绿豆多酚组成成分的影响及与其抗氧化活性的关系。结果表明:通过超高效液相色谱-质谱联用仪检测出46种多酚类化合物,筛选出44个差异代谢物,萌发前与萌发后绿豆芽的清除DPPH·能力分别为87.94%和96.24%,总抗氧化能力分别为28.56 U/mL和33.06 U/mL;最后通过KEGG查询得到筛选出的多酚类化合物参与了20个代谢通路。初步明确了绿豆中多酚类活性成分的物质基础,为绿豆的多酚类物质的合理利用与开发提供了参考。     

Inactivation of escherichia coli 0157:H7 and Salmonella on mung beans, alfalfa, and other seed types destined for sprout production by using an oxychloro-based sanitizer

The efficacy of a stabilized oxychloro-based food grade sanitizer to decontaminate seeds destined for sprout production has been evaluated. By using mung bean seeds as a model system, it was demonstrated that the sanitizer could be used to inactivate a five-strain cocktail of Escherichia coli O157:H7 or Salmonella introduced onto beans at 10(3) to 10(4) CFU/g. Salmonella was more tolerant to stabilized oxychloro than was E. coli O157:H7, with sanitizer levels of >150 and >50 ppm, respectively, being required to ensure pathogen-free sprouts. The decontamination efficacy was also found to be dependent on treatment time (>8 h optimal) and the seed-to-sanitizer ratio (>1:4 optimal). Stabilized oxychloro treatment did not exhibit phytotoxic effects, as germination and sprout yields were not significantly (P > 0.05) different as compared with untreated controls. Although human pathogens could be effectively eliminated from mung beans, the aerobic plate count of native microflora on sprouts grown from treated seed was not significantly (P > 0.05) different from the controls. The diversity of microbial populations (determined through 16S rRNA denaturing gradient gel electrophoresis analysis) associated with bean sprouts was not significantly affected by the sanitizer treatment. However, it was noted that Klebsiella and Herbasprillum (both common plant endophytes) were absent in sprouts derived from decontaminated seed but were present in control sprouts. When a further range of seed types was evaluated, it was found that alfalfa, cress, flax, and soybean could be decontaminated with the stabilized oxychloro sanitizer. However, the decontamination efficacy with other seed types was less consistent. It appears that the rate of seed germination and putative activity of sanitizer sequestering system(s), in addition to other factors, may limit the efficacy of the decontamination method.     

碾轧对绿豆淀粉的机械力化学效应

以绿豆淀粉为原料,通过扫描电镜(SEM)、偏光显微镜(PLM)、激光扫描共聚焦显微镜(CLSM)、X-射线衍射(XRD)、差示扫描量热仪(DSC)、快速黏度分析仪(RVA)等手段研究碾轧处理对淀粉结构和性质影响,探究其相互关系并揭示碾轧对绿豆淀粉机械力化学效应。结果表明,碾轧处理3~6 h时,淀粉无定型区和部分结晶区发生破坏,水溶指数、膨胀度、透光率增大,热焓减小。碾轧处理9 h时,淀粉内部发生重结晶,颗粒表面形成球状凸起,脐点区域直链淀粉聚集导致膨胀度、透光率、峰值黏度下降,水溶指数、热焓值、糊化温度增大。碾轧处理12~24 h时,淀粉的结晶区域发生显著破坏,颗粒严重变形,从而使淀粉水溶指数、透光率增大,膨胀度、热焓值减小。根据机械力化学相关理论推断淀粉颗粒内部依次经过了受力阶段、聚集阶段、团聚阶段。     

绿豆粉丝生产中淀粉粉团的流变特性研究

通过考察绿豆淀粉粉团在不同温度(20、30、40和50℃)、不同含水量(44%、47%、50%和53%)、不同淀粉糊含量(0%、12%、24%、36%和50%)、不同剪切速率(0.1、1、10、100和500s-1)和分别恒定剪切速率10s-1和100s-1从20℃到60℃连续升温扫描的条件下的流变特性,得出含水量47%,含淀粉糊量24%的粉团在温度为40℃,剪切速率在10～100s-1范围内最适合在绿豆粉丝生产中淀粉粉团的搅拌、输送和漏粉垂丝;同时用幂律方程、Cross方程和Herschel-Bulkley方程来描述粉团的流变特性,发现Cross方程比幂律方程拟合精度更高,而Herschel-Bulkley方程则适于描述纯淀粉浆团的流变状态和作为淀粉粉团在某些条件下流变状态描述的补充;并对其流变机理用颗粒在流场中的取向理论、胀容现象和触变流动现象进行了探讨。