Inactivation of Escherichia coli by Carbon Dioxide under Pressure

Thermal inactivation of Escherichia coli was studied under CO₂ pressures of 1.2, 2.5, and 5 MPa at 25, 35, and 45°C. Two phases were observed in the destruction curves. The earlier stage was characterized by a slow rate of inactivation, which increased sharply at the later stage. An increase of pressure and/or temperature enhanced the antimicrobial effects of CO₂ under pressure. The effects on cell structure were studied by scanning electron microscopy and the specific mechanism of action appeared to be related to enzyme inactivation.     

抗菌肽在大肠杆菌中融合表达的载体蛋白

抗菌肽是天然免疫系统的主要成分,能有效抵御病原微生物的侵害。目前其作为新型药物受到人们很大的关注,由于从天然资源中分离或化学合成成本较高,因此应用基因重组技术生产抗菌肽很迫切。本文主要讨论了抗菌肽在大肠杆菌中的融合表达、融合表达中常用的载体蛋白及新的融合载体SUMO（小泛素修饰因子）的主要性能。     

Detection of Escherichia coli O157 in bovine meat products in northern Italy

Tests for Escherichia coli and E. coli O157 were carried out on meat samples collected from randomly chosen stores throughout the city of Bologna and suburban areas. The samples consisted of 25 g of loose minced beef, sometimes already shaped into meatballs or hamburgers, some of which were mixed with vegetables. The meat was purchased from retail outlets, open market stalls, and supermarket chains during 25 sampling visits from October 2000 to December 2001. For E. coli detection, Tryptone soya broth (TSB) supplemented with novobiocin and C-EC agar were used. Immunomagnetic separation with SMAC-BCIG-CT agar and chromogenic E. coli O 157 agar, API 20E system and agglutination latex test were used to detect E. coli O157; Vero cell assay and polymerase chain reaction (PCR) were used to assess toxin production and the presence of virulence genes.

E. coli were detected in 45 (30.2%) of the 149 samples examined, mainly in the hamburger samples mixed with vegetables and in the loose minced beef. E. coli O157 was found in one sample of hamburger and two samples of hamburger mixed with vegetables (2%) collected from three different butcher's stores between July and October. All the strains of E. coli O157 and most cases of E. coli were found in meat from small retailers.

The three strains of E. coli O157 were positive for verocytotoxin production. PCR analysis revealed genes coding for vt2 and one strain possessed the gene for eae A. Chromogenic E. coli O157 agar was found to be more selective and differential, allowing easier identification of suspected colonies with mixed flora and producing less false-positive colonies.     

Microbiological Decontamination of Poultry Feed—Evaluation of Steam Conditioners

The microbial decontamination of chicken feed, obtained from a commercial pellet mill, was evaluated using a direct-fired steam conditioner (DFSC; APC System^™). The standard plate count of the feeds before (mash) and after (pellets) conditioning ranged from 65×10⁴ to 83×10⁵ colony forming units (CFU) g⁻¹ and from 91×10¹ to 92×10³ CFU g⁻¹, respectively. The incidence of Escherichia coli , Salmonella and Listeria in the feeds before conditioning was 61·7, 8·3 and 27·1%, respectively. Following conditioning these levels were reduced to 1·7, 1·7 and 0%, respectively. Species of Listeria and Salmonella identified included L monocytogenes , L innocua and S agona , S ohio , S heidelberg , S senftenberg , S tallahasse and S braenderup , respectively. Compared with a conventional, indirect-fired boiler-generated-steam conditioner (IFSC) the direct-fired steam conditioner proved superior in regards to pathogen decontamination; no E coli , Salmonella or Listeria were recovered from mash lots positive for these microorganisms. However, with the IFSC system, both E coli and L monocytogens were recovered at levels of 11·1 and 5·6%, respectively.     

利用EHEC琼脂平板从食物中分离检测产志贺毒素大肠杆菌

为利用肠出血性大肠杆菌(EnterohaemorrhagicEscherichiacoli,EHEC)琼脂平板,从食物中分离、检测产志贺毒素大肠杆菌(shigatoxin-producingEscherichiacoli,STEC),采用STEC毒力基因(hlyA基因)检测溶血素的产生,建立了一种利用EHEC琼脂平板快速、准确地从食物中分离、检测STEC的方法。该方法可检测STEC不同血清O157∶H7、O26、O111。     

大肠杆菌耐酸分子机制研究进展

具有耐强酸能力的大肠杆菌在酸性胁迫条件下更易生存。这主要是由于其具有葡萄糖-阻碍耐酸系统、氨基酸依赖型耐酸系统、伴侣蛋白抗酸作用以及保持膜电荷稳定等耐酸机制。了解大肠杆菌的这些耐酸分子机制,可以为食品加工工业控制大肠杆菌的污染提供新的认识,也对一些食源性致病菌的临床预防和治疗具有积极的作用。本文就大肠杆菌耐酸分子机制的研究进行综述。     

基于特征基因的不同类型致泻大肠埃希氏菌的分子鉴定

致泻大肠埃希氏菌是引起人体以腹泻症状为主的常见病原菌。将传统微生物学方法和分子鉴定方法相结合鉴定致泻大肠埃希氏菌。根据聚合酶链式反应(polymerase chain reaction,PCR)扩增和测序建立5种不同类型致泻大肠埃希氏菌的分子鉴定标准,并对试验过程进行安全性评价,最后通过检测鲜肉中肠道集聚性大肠埃希氏菌验证此方法的适用性。结果显示5种致泻大肠埃希氏菌标准菌株的特征基因都有明显的条带,且与该特征基因的产物条带大小一致。同时,胶回收测序的结果与特征基因的序列一致性为100%,可利用PCR和测序鉴定5种致泻大肠埃希氏菌;在安全性评价试验中,利用细菌裂解液进行涂板,培养后均未发现菌落生成,因此没有生物安全隐患;肉样中的检测结果显示,有astA基因的目的条带,且测序结果与此基因序列一致,说明此方法能够准确鉴定出鲜肉中肠道集聚性大肠埃希氏菌。成功建立5种不同类型大肠埃希氏菌的分子鉴定方法,评价试验过程的安全性,并准确鉴定出肉样中的肠道集聚性大肠埃希氏菌,为实验室相关标准的制定和修订提供理论依据和有益借鉴。     

新科斯糖对肠道菌体外生长影响的研究

研究新科斯糖对大肠杆菌和双歧杆菌体外生长的影响。结果表明,以葡萄糖和普通低聚果糖为对照,大肠杆菌在新科斯糖上的生长相对弱一些,但差异不显著(p>0.05);双歧杆菌能很好的利用新科斯糖在体外生长,是利用普通低聚果糖增殖效果的1.4倍,葡萄糖的2倍。     

生物合成对香豆酸大肠杆菌工程菌的构建

对香豆酸是一种具有预防心血管疾病、抗氧化和抗菌消炎等生物活性的酚类物质,同时,它也是高价值苯丙烷类保健营养品(如白藜芦醇)的前体。本研究希望创制出一种生物合成对香豆酸的方法,以缓解对香豆酸的供求问题。把带有粘红酵母(Rhodotorula glutinis)酪氨酸解氨酶(Rg TAL)基因的组成型表达载体转化大肠杆菌ATCC31884,并通过PCR鉴定后,获得重组菌株。对重组菌株进行发酵培养,对发酵液进行高效液相色谱检测,确定工程菌具有生物合成对香豆酸的能力。随后通过优化L-酪氨酸的添加量和工程菌的发酵时间,最终确定,底物L-酪氨酸的添加量为0.5 m M,发酵时间为36 h,检测发酵液中的对香豆酸含量最高,为161.23 mg/L。这表明,Rgtal基因在重组大肠杆菌中成功得到了表达,并且能利用自身代谢和外源添加的L-酪氨酸生物合成对香豆酸。     

大肠杆菌cdd和thrA基因的敲除及其对胞苷积累量的影响

以大肠杆菌AU39作为出发菌株,通过基因工程手段对其进行改造,旨在提高胞苷产量。首先,通过Red重组系统敲除了大肠杆菌AU39基因组上的胞苷脱氨酶基因cdd,阻断了胞苷的分解代谢;敲除了E．coliAU39（Acrid）基因组上的高丝氨酸脱氢酶基因thrA,阻断天冬氨酸向高丝氨酸的代谢途径。然后,分别对不同基因缺失菌株进行培养发酵,与出发菌株E．coliAU39相比,两株突变株都有不同程度的胞苷积累,E．coliAU39（Acdd）与E．coliAU39（AcddAthrA）的胞苷产量提高了1．25倍与1．6倍,而尿苷产量均相对有所降低。     

食源性大肠杆菌快速检测技术研究进展

大肠杆菌又称大肠埃希氏菌(Escherichia coli),其中有少数血清型可引起食源性疾病,对人类生命健康产生很大危害。因此,研究快速、灵敏、特异的食源性大肠杆菌检测方法对有效保障食品安全有着重要意义。本文对食源性大肠杆菌快速检测技术的研究进展进行了综述。      

2007-2009年欧盟肠出血性大肠杆菌监测简介

2007—2009年欧盟平均每年报告3 213例肠出血性大肠杆菌病例,平均每年溶血性尿毒综合征患者185例,血清型以肠出血性大肠杆菌O157为主。食品与牛粪便中分离的肠出血性大肠杆菌血清型以O157为主。2007—2009年鲜牛肉肠出血性大肠杆菌O157分离率分别为0.1%、0.1%和0.7%,牛粪便中肠出血性大肠杆菌O157分离率分别为2.9%、0.5%和2.7%。     

Bifidobacteria as indicators of faecal contamination in meat and meat products: detection, determination of origin and comparison with Escherichia coli

Faecal contamination of meat and meat products and its origin, whether human or animal, was determined by using the presence of bifidobacteria as an indicator. Enrichment of samples in Beerens liquid selective medium was followed by spreading onto Columbia agar containing paromomycin. In comparison with the detection of Escherichia coli (E. coli) the following results were obtained from 50 samples: E. coli. +; Bifidobacterium. +: 38; E. coli −; Bifidobacterium −: 9; E. coli, +; Bifidobacterium. −: 2; E. coli, −; Bifidobacterium. +: 1. From 39 positives samples, 50 strains of bifidobacteria were isolated. Two were of human origin, 48 of animal origin.     

高强脉冲电场对食品中大肠埃希氏菌基因水平传递的影响

目的:探讨高强脉冲电场对大肠埃希氏菌的基因获得及水平传递中的作用。方法:采用接合转移试验,研究了PEF强电场作用对大肠埃希氏菌基因水平传递的影响,并以未经处理的标准菌株为对照,通过PCR方法进行基因检测分析。结果:所有经PEF处理的菌株均由正常的接合转移率42.4%～68.5%上升到100%。PCR检测结果显示,所得接合子中int基因的扩增产物大小仍为923bp,与正常未处理int基因大小一致。结论:PEF处理使得细菌接合转移率明显提高,PEF处理对传递到接合子中的基因未能产生影响。     

18型人乳头瘤病毒E6蛋白截短体(HPV18 E6*)的基因克隆与表达

目的 构建HPV18 E6*原核重组表达质粒,并优化其蛋白表达条件.方法 以人宫颈癌HeLa细胞cDNA为模版,PCR扩增HPV18 E6*基因,构建重组表达载体HPV18 E6*-pET28a,在大肠杆菌BL21 (DE3)中诱导表达重组蛋白,并优化其表达条件,SDS-PAGE检测重组蛋白表达状况.结果 成功构建了重组表达载体;37℃表达,HPV18 E6*以不溶包涵体为主,表达量占菌体总蛋白的20％左右;15℃主要为可溶形式;包涵体变复性获得纯蛋白.结论 HPV18 E6*蛋白的高效表达为研究其蛋白作用机制及为研究预防和治疗子宫癌奠定基础.     

二氧化氯(ClO2)对葡萄表面大肠杆菌杀灭效果的研究

用CiO2溶液浸泡已接种大肠杆菌的葡萄,观察处理时间、ClO2溶液体积与葡萄质量之比(v/m)、ClO2浓度对杀菌效果的影响.结果表明:处理时间从5min延长到10min杀菌效果上升2.35±0.18log,但进一步延长处理时间上升趋势变缓;v/m为2时的杀菌效果低于其为3、4、5时的杀菌效果,后3者之间无显著性差异(P＞0.05);ClO2浓度在很大程度上影响着杀菌效果,ClO2浓度增加杀菌效果明显提高.     

食品中大肠杆菌的快速检测方法

大肠杆菌是人及各种动物肠道中的正常寄居菌,食物或水中大肠杆菌的检出意味着直接或间接的近期粪便污染。大肠杆菌作为饮水、食品等的粪源性污染卫生细菌学指标;而且他在外界存活时间与一些主要肠道病原菌相近,它的出现也可能预示某些肠道病原菌（如沙门氏菌、志贺氏菌）的存在。大肠杆菌是国际上公认的卫生监测指示菌,因此大肠杆菌的检测技术显得十分重要,相应出现了大量的大肠杆菌的各种检测方法。     

大肠杆菌工程菌产木聚糖酶工艺优化及酶学性质研究

对大肠杆菌工程菌产木聚糖酶的工艺进行优化,并对木聚糖酶的酶学性质进行研究。确定了提取工艺路线,即依次经过细胞破碎(高速珠磨法)、固液分离及除菌(中空纤维膜过滤)、超滤浓缩及烘干包衣步骤后得到肠溶木聚糖酶产品。该产品热稳定性好,在pH 2.5磷酸盐缓冲液中释放度为5.0%,在pH 5.5磷酸盐缓冲液中释放度为98.5%,产品外观光泽性好,能够很好地满足饲料酶市场要求。该木聚糖酶的最适pH值为6.4,最适温度为55℃。该酶具有较好的热稳定性,含水分17%的酶在90℃条件下2.5min仅失活13.6%。对木聚糖酶降解产物进行检测表明该木聚糖酶是一种内切酶。