环境因素对大肠杆菌REP-PCR指纹图谱稳定性影响研究

大肠杆菌作为粪便污染指示生物,其DNA指纹图谱稳定性是制约微生物污染源识别技术发展的瓶颈之一.本研究采用绿色荧光蛋白基因(GFP)标记后的Escherichia coli K12(K12GFP)为受试菌株,利用REP-PCR方法,对大肠杆菌在不同温度、光照、盐度条件下其DNA指纹图谱的稳定性进行了研究.结果表明,绿色荧...     

聚合酶链式反应(PCR)用于检测环境水体指示菌的研究

对聚合酶链式反应（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ，ＰＣＲ）快速检测水体指示菌－大肠菌群（ｔｏｔａｌ ｃｏｌｉｆｏｒｍ）和大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ）进行了研究。采用的两对引物分别扩增了ＬａｃＺ基因的约２６０ｂｐ和ＵｄｉＡ基因的约１５０ｂｐＤＮＡ片段。引物Ⅰ和引物Ⅱ最低能分别扩增１０＾－６μｇ或１０＾－８μｇ基因组ＤＮＡ。整个检测工作在采集水样后５－６ｈ内完成。与     

Antibiotic resistance and virulence factors of Escherichia coli from eagles and goshawks

One of the major global problems in medicine is microbial resistance to antibiotics (antimicrobial resistance) and this has become an increasingly frequent research topic. This study focuses on antimicrobial resistance, phylogenetic and genetic characterization of Escherichia coli from wild birds: ten isolates from eagles (Aquila chrysaetos), nine from goshawks (Accipiter gentilis) and 24 from broilers in the Slovak Republic. Twenty-two strains with presence of int1 gene were selected and examined for the presence or absence of transposon gene (tn3), genes of antibiotic resistance and virulence factors. We detected sequence type (ST) in eagles ST 442 with genes iss, papC, iutA, cvaC, tsh, fyuA, iroN, kps, feoB, sitA, irp2, ireA for virulence factors and tetA, sul1, sul2, dfrA, aadA for antibiotic resistance; in goshawks ST 1011 with iss, papC, fyuA, iroN, feoB, sitA and qnrS1, tetA, sul1, sul2, dfrA, aadA, respectively. These ST types have been found in humans too and should be evaluated further for possible zoonotic potential and transfer of resistance genes from the environment.     

Antibiotic resistance,virulence factors and biofilm formation ability in Escherichia coli strains isolated from chicken meat and wildlife in the Czech Republic

Attachment of pathogenic bacteria to food contact surfaces and the subsequent biofilm formation represent a serious threat for the food industry, since these bacteria are more resistant to antimicrobials or possess more virulence factors. The main aim of this study was to investigate the correlation between antibiotic resistance against 13 antibiotics, distribution of 10 virulence factors and biofilm formation in 105 Escherichia coli strains according to their origin. The high prevalence of antibiotic resistance that we have found in wildlife isolates could be acquired by horizontal transfer of resistance genes from human or domestic or farm animals. Consequently, these commensal bacteria might serve as indicator of antimicrobial usage for human and veterinary purposes in the Czech Republic. Further, 46 out of 66 resistant isolates (70%) were able to form biofilm and we found out statistically significant correlation between prevalence of antibiotic resistance and biofilm formation ability. The highest prevalence of antibiotic resistance was observed in weak biofilm producers. Biofilm formation was not statistically associated with any virulence determinant. However, we confirmed the correlation between prevalence of virulence factors and host origin. Chicken isolates possessed more virulence factors (66%), than isolates from wildlife (37%). We can conclude that the potential spread of antibiotic resistance pattern via the food chain is of high concern for public health. Even more, alarming is that E. coli isolates remain pathogenic potential with ability to form biofilm and these bacteria may persist during food processing and consequently lead to greater risks of food contamination.     

大肠杆菌JM105中硝酸盐还原酶的制备及其性质

近年来,硝酸盐和亚硝酸盐的污染已引起普遍关注。目前,蔬菜和粮食的NO3-含量过高主要由于农药和化肥的使用、工业废水或生活污水灌溉等,食品类主要来自腌制食品和食品添加剂或防腐剂等。另外,向乳制品或食品中参碱、食盐、化肥、脏水、碱性水等也是硝酸盐污染的来源之一。因此,迫切需要快速、简便、可用于现场的硝酸盐检测方法。这就需仰赖生物酶方法,而生物酶方法的关键是酶制剂的制备和检测方法的建立。本研究通过筛选、厌氧和硝酸盐诱导培养、超声波细胞破碎和差速离心提取等方法,从大肠杆菌(Escherichia coli)JM105细胞膜中制备了硝酸盐还原酶并对其性质进行了研究。结果表明:从大肠杆菌JM105中制得的酶制剂活力很高,且在酶过量的情况下可将NO3-完全转化为NO2-,在用磷酸缓冲液清洗并冷冻保藏过夜后不含有亚硝酸盐还原酶。在加入黄素单核苷酸辅酶(FMN)后,该酶的活力可提高64%,比活力达0.42U·mg-1蛋白。该酶十分稳定,在40℃下24h活力无影响,在浓度为1mmol·L-1的Cu2 、Fe3 、Ca2 、Zn2 、Mg2 和Mo6 存在下,其活力亦不改变。因此,该酶可用于测定食品、蔬菜和环境中硝酸盐的含量。     

FECAL‐INDICATOR BACTERIA IN STREAMS ALONG A GRADIENT OF RESIDENTIAL DEVELOPMENT1

ABSTRACT: Fecal‐indicator bacteria were sampled at 14 stream sites in Anchorage, Alaska, USA, as part of a study to determine the effects of urbanization on water quality. Population density in the subbasins sampled ranged from zero to 1,750 persons per square kilometer. Higher concentrations of fecal‐coliform, E. coli, and enterococci bacteria were measured at the most urbanized sites. Although fecal‐indicator bacteria concentrations were higher in summer than in winter, seasonal differences in bacteria concentrations generally were not significant. Areas served by sewer systems had significantly higher fecal‐indicator bacteria concentrations than did areas served by septic systems. The areas served by sewer systems also had storm drains that discharged directly to the streams, whereas storm sewers were not present in the areas served by septic systems. Fecal‐indicator bacteria concentrations were highly variable over a two‐day period of stable streamflow, which may have implications for testing of compliance to water‐quality standards.     

Photo- and electro-catalysis for solar disinfection of Escherichia coli-contaminated water using Ag–TiO2/graphite

A titanium dioxide film on a graphite substrate was synthesized by chemical bath deposition from TiCl₄ as precursor and with the surfactant cetyl trimethyl ammonium bromide as a linking and assembling agent. Silver was loaded on the TiO₂ film by electrodeposition at 0.025?A. Water contaminated with Escherichia coli was disinfected under sunlight irradiation by photolysis (Lys), photocatalysis (PC), photoelectrocatalysis (PEC), and electrocatalysis (EC). The highest rate constant, k, was achieved with EC; k was 5.1?×?10^?2 colony forming units (CFU) mL^?1?min^?1. However, auto-oxidation of Ag occurred during EC and PEC. Meanwhile, the rate constant of disinfection by means of PC was lower than EC and PEC, and k was 3.82?×?10^?2 CFU?mL^?1?min^?1. Nevertheless, the auto-oxidation of Ag in the Ag–TiO₂/graphite tablet did not occur during the disinfection process.     

拟南芥硫酯酶基因(atfata)在大肠杆菌中的表达及其对游离脂肪酸合成的影响

硫酯酶催化脂酰ACP(酰基载体蛋白)水解生成游离脂肪酸和ACP载体蛋白,能够解除脂肪酸合成与磷脂合成的偶联,是获得游离脂肪酸的关键基因.将拟南芥的硫酯酶基因atfata克隆到原核表达载体pET30a上,在大肠杆菌BL21(DE3)中成功表达了His-tag融合蛋白,经SDS-PAGE检测获得特异性表达条带.对该重组菌株进行摇瓶发酵实验,GC-MS检测表明其与对照菌株脂肪酸组成并未发生变化;但其脂肪酸产量达到232.06 mg/L,比对照菌株提高了70%;同时胞外游离脂肪酸产量达到了33 mg/L,占总脂肪酸含量的15%,是野生菌株胞外脂肪酸产量的7倍.图3表2参17     

饮用水中典型微生物消毒过程中消毒副产物的生成规律

以饮用水中典型微生物--大肠杆菌（Escherichia coli）为试验对象,研究pH值、氯化时间、氯投量及细菌浓度对大肠杆菌在氯化消毒过程中生成消毒副产物（DBPs）的影响,并分析何种氯化条件下,DBPs控制效果最佳.研究表明：随氯投量增加,二氯乙腈（DCAN）呈先上升后下降趋势;随氯化时间延长,三氯丙酮（1,1,1-TCP）和DCAN先增加后减少;在pH值从5升高到9时,1,1,1-DCP、三氯硝基甲烷（TCNM）、二氯乙酸（DCAA）和三氯乙酸（TCAA）持续降低;细菌污染水源事件在近年常有报道,当水源水中细菌浓度增加时,饮用水中三氯甲烷（TCM）、TCNM、DCAA和TCAA浓度增加,但DCAN、三氯乙腈（TCAN）、二氯丙酮（1,1-DCP）和1,1,1-TCP不一定增加.为了达到低毒性的目的,氯投量浓度不宜太高,同时控制氯化时间为6h和pH>8.     

大肠杆菌配合植酸对铀的吸附

研究了用植酸处理后大肠杆菌对铀进行吸附,考察了pH、初始浓度、反应时间以及菌液投加浓度对吸附效果的影响。实验结果表明,植酸能够提高大肠杆菌菌悬液对铀的吸附效果。植酸处理后的大肠杆菌能有效结合UO22+,在pH=4.0时,菌液投加浓度为90%时,吸附效果最好,吸附300 min达到平衡,吸附率为90%。大肠杆菌配合植酸吸附铀前后电镜图及EDX表明,反应能使铀酰离子络合沉淀,沉积在菌体表面。