大肠埃希菌和肺炎克雷伯菌临床分离株的耐药性分析

目的分析临床分离的大肠埃希菌和肺炎克雷伯菌的分布情况及耐药性。方法收集吉林大学中日联谊医院临床标本中分离的221株大肠埃希菌和152株肺炎克雷伯菌,采用纸片扩散法检测抗菌素的敏感性;双纸片协同试验和纸片表型确证试验筛选并确证产超广谱β-内酰胺酶的菌株;按照美国国家临床实验室标准化委员会(NCCLS)2005年版的标准判断结果。结果大肠埃希菌和肺炎克雷伯菌对亚胺培南和美罗培南的敏感性最高,均达100%;对头孢他啶、哌拉西林/他唑巴坦和头孢西丁的敏感性也较高,均大于70%,而对氨苄西林的耐药性均大于90%。共检出产超广谱β-内酰胺酶的大肠埃希菌118株,检出率为53.4%;肺炎克雷伯菌21株,检出率为13.8%;产超广谱β-内酰胺酶的两种菌株与非产超广谱β-内酰胺酶的同种菌株相比,对抗菌素的耐药性均明显增加。结论大肠埃希菌和肺炎克雷伯菌仍是产超广谱的β-内酰胺酶的主要菌株,且对常用抗菌素产生了较高的耐药性。     

内蒙古通辽地区犊牛腹泻大肠埃希菌毒力基因检测及耐药性分析

目的 对内蒙古通辽地区犊牛腹泻大肠埃希菌(E. coli)的毒力基因进行检测,并分析其耐药性及耐药基因分布情况。方法 采用药敏纸片法检测从腹泻犊牛粪便样品分离鉴定的82株致病性E. coli对13种抗菌药物的敏感性,PCR法检测其13种毒力基因和12种耐药基因携带情况,并对其进行系统进化分群。结果 82株致病性E. coli中48.78%(40/82)、31.71%(26/82)、14.63%(12/82)和4.88%(4/82)分别属于A、B1、B2和D群,优势群系为A群(48.78%,40/82);共检测到11种毒力基因,未检测到tsh和LT1,其中irp2、fyuA、eaeA和STb的检出率较高,分别为79.27%(65/82)、63.41%(52/82)、53.66%(44/82)和50%(41/82),其他毒力基因检出率均低于50%。82株致病性E. coli对13种抗菌药物的耐药情况严重,其中对四环素、多西环素和阿莫西林的耐药性普遍较高,耐药菌株占比分别为100%(82/82)、97.56%(80/82)和90.24%(74/82);全部菌株表现出多重耐药,耐8种抗菌药物的...     

吡哌酸一季度需求同比增六成半

据统计，今年一季度的吡哌酸需求强劲，同比净增六成半以上． 吡哌酸为喹诺酮类抗菌药，通过作用于细菌DNA旋转酶，干扰细菌DNA的合成，从而导致细菌死亡，对革兰阴性杆菌如大肠埃希菌、肺炎克雷伯菌、产气肠杆菌等具抗菌作用，疗效显著且价格十分低廉．     

吡哌酸需求强劲

据统计，2007年一季度的吡哌酸需求强劲，同比净增六成半以上． 吡哌酸为喹诺酮类抗菌药，通过作用于细菌DNA旋转酶，干扰细菌DNA的合成，从而导致细菌死亡，对革兰阴性杆菌如大肠埃希菌、肺炎克雷伯菌、产气肠杆菌等具抗菌作用，疗效显著且价格十分低廉．     

臭氧对水中大肠埃希氏菌和肺炎性军团杆菌的失活作用

臭氧是自然界中存在的强氧化剂。目前在许多发达国家和我国部分地区都有用作生活用水处理的最后消毒手段。本文利用近年来国外发表的文献资料和实验结果,简明扼要地介绍和讨论臭氧对大肠埃希氏菌(E Coli)和肺炎性军团杆菌(Legionella pneumophila)的失活作用。     

阴沟肠杆菌对石油烃污染物的降解特性

考察了阴沟肠杆菌E．cloacae对石油烃代表物十六烷及菲的降解特性,分析了菌在各碳源中的生长动力学曲线及各碳源的降解动力学曲线。结果表明E.cloacae在十六烷、菲、十六烷和菲的混合物这三种碳源中都能较好地生长,生长速率较高,生长过程中都有二次生长现象;E．cloacae对葡萄糖、十六烷、菲、以及十六烷和菲混合物中所含菲在1天内平均降解速率大小关系为十六烷〉葡萄糖〉菲（十六烷与菲混合）〉菲,E．cloacae对十六烷具有很强的降解能力,1天内其对十六烷的平均降解速率是其对葡萄糖的2．4倍,是对单独菲及混合菲的6200倍。最终结果表明E．cloacae是一种高效的石油烃降解菌。     

γ-氧化铁/山梨酸/银-二氧化钛的制备及抗菌性能

采用溶胶凝胶-水热法制备了γ-氧化铁/山梨酸/银-二氧化钛催化剂。以可见光驱动下对大肠埃希氏菌的对数去除率为评价指标,获得了催化剂的最佳制备参数：n（银）/n（钛）=0.03、n（山梨酸）/n（钛）=0.2、水热温度为160 ℃、水热时间为12 h。利用X射线衍射（XRD）、X射线光电子能谱（XPS）、BET法比表面积（BET）、高倍扫描电镜（FESEM）等表征方法对其结构、表面特性、光学性能和磁性性能进行分析。催化剂的物化性能优异：构成为纯锐钛矿二氧化钛、顺磁性γ-氧化铁和单质银,山梨酸以单齿方式与二氧化钛结合;外观为类球形构成的比表面积为125.726 m²/g的介孔结构;可响应200~800 nm全波段光;是饱和磁化强度为10.478 A·m²/kg的超顺磁材料。自由基淬灭试验确定活性氧（ROS）影响抗菌活性的顺序从大到小依次为·OH、O₂^·-、h⁺。自由基、游离银离子和山梨酸构成协同抗菌体系,使催化剂在可见光和黑暗条件下均具有良好的抗菌活性,对数去除率分别为5.91和2.54。     

大肠埃希菌对人结肠癌细胞RKO产生β防御素-3的影响

<正>β防御素是天然抗菌肽家族的重要成员,同时也是机体先天性防御屏障的重要组成部分,对细菌、真菌和病毒入侵机体具有重要的防御作用~[1-2]。β防御素-3通常受外界环境的刺激而产生~[3],在人体上皮细胞和黏膜组织中广泛表达,具有广谱的抗微生物活性,能促进伤口愈合,发挥免疫调节     

肠杆菌X-11对石油烃污染物的降解特性

考察了肠杆菌x-11对石油烃代表物十六烷及菲的降解特性,分析了菌在各碳源中的生长动力学曲线及各碳源的降解动力学曲线.结果表明肠杆菌X-11在十六烷、菲、十六烷与菲混合物这3种碳源中都能较好地生长,都能达到较高的生长速率,生长过程中都有二次生长现象;肠杆菌X-11对葡萄糖、十六烷、菲、以及十六烷与菲混合物中所含菲的1天内平均降解速率大小关系为:十六烷＞葡萄糖＞菲(十六烷与菲混合)＞菲,肠杆菌X-11对十六烷具有很强的降解能力.其对十六烷的1天内平均降解速率是其对葡萄糖的2.4倍,是对单独菲及混合菲的6 200倍.     

Virulence Factors of Enteric Pathogenic Escherichia coli: A Review

Escherichia coli are remarkably versatile microorganisms and important members of the normal intestinal microbiota of humans and animals. This harmless commensal organism can acquire a mixture of comprehensive mobile genetic elements that contain genes encoding virulence factors, becoming an emerging human pathogen capable of causing a broad spectrum of intestinal and extraintestinal diseases. Nine definite enteric E. coli pathotypes have been well characterized, causing diseases ranging from various gastrointestinal disorders to urinary tract infections. These pathotypes employ many virulence factors and effectors subverting the functions of host cells to mediate their virulence and pathogenesis. This review summarizes new developments in our understanding of diverse virulence factors associated with encoding genes used by different pathotypes of enteric pathogenic E. coli to cause intestinal and extraintestinal diseases in humans.     

Correlation between CRISPR Loci Diversity in Three Enterobacterial Taxa

CRISPR-Cas is an adaptive immunity system of prokaryotes, composed of CRISPR arrays and the associated proteins. The successive addition of spacer sequences in the CRISPR array has made the system a valuable molecular marker, with multiple applications. Due to the high degree of polymorphism of the CRISPR loci, their comparison in bacteria from various sources may provide insights into the evolution and spread of the CRISPR-Cas systems. The aim of this study was to establish a correlation between the enterobacterial CRISPR loci, the sequence of direct repeats (DR), and the number of spacer units, along with the geographical origin and collection source. For this purpose, 3474 genomes containing CRISPR loci from the CRISPRCasdb of Salmonella enterica, Escherichia coli, and Klebsiella pneumoniae were analyzed, and the information regarding the isolates was recorded from the NCBI database. The most prevalent was the I-E CRISPR-Cas system in all three studied taxa. E. coli also presents the I-F type, but in a much lesser percentage. The systems found in K. pneumoniae can be classified into I-E and I-E*. The I-E and I-F systems have two CRISPR loci, while I-E* has only one locus upstream of the Cas cluster. PCR primers have been developed in this study for each CRISPR locus. Distinct clustering was not evident, but statistically significant relationships occurred between the different CRISPR loci and the number of spacer units. For each of the queried taxa, the number of spacers was significantly different (p < 0.01) by origin (Africa, Asia, Australia and Oceania, Europe, North America, and South America) but was not linked to the isolation source type (human, animal, plant, food, or laboratory strains).     

Presence of β-Lactamase-producing Enterobacterales and Salmonella Isolates in Marine Mammals

Marine mammals have been described as sentinels of the health of marine ecosystems. Therefore, the aim of this study was to investigate (i) the presence of extended-spectrum β-lactamase (ESBL)- and AmpC-producing Enterobacterales, which comprise several bacterial families important to the healthcare sector, as well as (ii) the presence of Salmonella in these coastal animals. The antimicrobial resistance pheno- and genotypes, as well as biocide susceptibility of Enterobacterales isolated from stranded marine mammals, were determined prior to their rehabilitation. All E. coli isolates (n = 27) were screened for virulence genes via DNA-based microarray, and twelve selected E. coli isolates were analyzed by whole-genome sequencing. Seventy-one percent of the Enterobacterales isolates exhibited a multidrug-resistant (MDR) pheno- and genotype. The gene bla_CMY (n = 51) was the predominant β-lactamase gene. In addition, bla_TEM-1 (n = 38), bla_SHV-33 (n = 8), bla_CTX-M-15 (n = 7), bla_OXA-1 (n = 7), bla_SHV-11 (n = 3), and bla_DHA-1 (n = 2) were detected. The most prevalent non-β-lactamase genes were sul2 (n = 38), strA (n = 34), strB (n = 34), and tet(A) (n = 34). Escherichia coli isolates belonging to the pandemic sequence types (STs) ST38, ST167, and ST648 were identified. Among Salmonella isolates (n = 18), S. Havana was the most prevalent serotype. The present study revealed a high prevalence of MDR bacteria and the presence of pandemic high-risk clones, both of which are indicators of anthropogenic antimicrobial pollution, in marine mammals.     

Production of 2,3-Butanediol by Klebsiella Pneumoniae Using Glucose and Ammonium Phosphate

The production of 2,3-butanediol by Klebsiella pneumoniae from glucose supplemented with different salts was studied. A suitable medium composition was defined by response surface experiments. In a medium containing glucose and （NH4）2HPO4, the strain could convert 137.0g of glucose into 52.4g of 2,3-butanediol and 8.4g of acetoin in shaking flasks. The diol yield amounted to 90% of its theoretical value and the productivity was 1-1.5g.L^-1.h^-1. In fed-batch fermentation, the yield and productivity of diol were further enhanced by maintaining the pH at 6.0. Up to 92.4g of 2,3-butanediol and 13.1g of acetoin per liter were obtained from 215.0g of glucose per liter. The diol yield reached 98% of its theoretical value and the productivity was up to 2.1g.L^-1.h^-1.     

Functional Dissection of P1 Bacteriophage Holin-like Proteins Reveals the Biological Sense of P1 Lytic System Complexity

P1 is a model temperate myovirus. It infects different Enterobacteriaceae and can develop lytically or form lysogens. Only some P1 adaptation strategies to propagate in different hosts are known. An atypical feature of P1 is the number and organization of cell lysis-associated genes. In addition to SAR-endolysin Lyz, holin LydA, and antiholin LydB, P1 encodes other predicted holins, LydC and LydD. LydD is encoded by the same operon as Lyz, LydA and LydB are encoded by an unlinked operon, and LydC is encoded by an operon preceding the lydA gene. By analyzing the phenotypes of P1 mutants in known or predicted holin genes, we show that all the products of these genes cooperate with the P1 SAR-endolysin in cell lysis and that LydD is a pinholin. The contributions of holins/pinholins to cell lysis by P1 appear to vary depending on the host of P1 and the bacterial growth conditions. The pattern of morphological transitions characteristic of SAR-endolysin–pinholin action dominates during lysis by wild-type P1, but in the case of lydC lydD mutant it changes to that characteristic of classical endolysin-pinholin action. We postulate that the complex lytic system facilitates P1 adaptation to various hosts and their growth conditions.     

维生素C和E对Klebsiella pneumoniae合成1,3-丙二醇的调控

通过外源添加还原剂的方式调控细胞内NADH/NAD再生系统的状态,研究了40~150 mg/L VC及20~100 mg/L VE对Klebsiella penumoniae合成1,3-丙二醇的影响,发现外源添加150 mg/L VC或30 mg/L VE均可使1,3-PD合成浓度提高20%~30%;但同时也提高了某些副产物的合成浓度,对代谢流分布的调控作用不明显;1,3-丙二醇得率稍有提高但不显著. 提高1,3-PD得率宜从代谢节点(丙酮酸)通量调节方面考虑.     

木瓜凝乳蛋白酶基因的克隆及在大肠杆菌中的表达

目的构建木瓜凝乳蛋白酶(CP)的表达载体,并在大肠杆菌中表达。方法从生番木瓜中提取mRNA后进行反转录,得到木瓜凝乳蛋白酶的cDNA,PCR产物克隆到pET22-b(+)载体中,重组质粒转化至大肠杆菌BL21,进行诱导表达及表达产物的检测。结果表达产物的SDS-PAGE显示单一条带,相对分子质量约为26000。Western blot表明表达产物可与抗木瓜凝乳蛋白酶特异性抗体结合。结论CP基因已成功在大肠杆菌中克隆表达,为进一步纯化和动物实验奠定了基础。     

重组Echistatin融合基因工程菌高密度发酵工艺优化

目的优化大肠杆菌表达重组蛇毒锯鳞蝰素(Echistatin,Ecs)融合基因工程菌的发酵工艺。方法在15L发酵罐内,研究培养基、培养条件和诱导时间对工程菌生长和目的蛋白表达的影响,并考察工程菌中重组质粒的遗传稳定性。结果工程菌在pH7·4的2×YT培养基中诱导4h,菌体湿重可达75g/L,目的蛋白表达量约占总蛋白的35%,所构建的重组质粒在BL21宿主菌中传代稳定。结论优化了Ecs融合基因工程菌的发酵和表达条件,为规模化生产奠定了基础。     

Molecular Epidemiology of Enteroaggregative Escherichia coli (EAEC) Isolates of Hospitalized Children from Bolivia Reveal High Heterogeneity and Multidrug-Resistance

Enteroaggregative Escherichia coli (EAEC) is an emerging pathogen frequently associated with acute diarrhea in children and travelers to endemic regions. EAEC was found the most prevalent bacterial diarrheal pathogen from hospitalized Bolivian children less than five years of age with acute diarrhea from 2007 to 2010. Here, we further characterized the epidemiology of EAEC infection, virulence genes, and antimicrobial susceptibility of EAEC isolated from 414 diarrheal and 74 non-diarrheal cases. EAEC isolates were collected and subjected to a PCR-based virulence gene screening of seven virulence genes and a phenotypic resistance test to nine different antimicrobials. Our results showed that atypical EAEC (a-EAEC, AggR-negative) was significantly associated with diarrhea (OR, 1.62, 95% CI, 1.25 to 2.09, p < 0.001) in contrast to typical EAEC (t-EAEC, AggR-positive). EAEC infection was most prevalent among children between 7–12 months of age. The number of cases exhibited a biannual cycle with a major peak during the transition from warm to cold (April–June). Both typical and a-EAEC infections were graded as equally severe; however, t-EAEC harbored more virulence genes. aap, irp2 and pic were the most prevalent genes. Surprisingly, we detected 60% and 52.6% of multidrug resistance (MDR) EAEC among diarrheal and non-diarrheal cases. Resistance to ampicillin, sulfonamides, and tetracyclines was most common, being the corresponding antibiotics, the ones that are frequently used in Bolivia. Our work is the first study that provides comprehensive information on the high heterogenicity of virulence genes in t-EAEC and a- EAEC and the large prevalence of MDR EAEC in Bolivia.