创伤弧菌抗菌药物药敏实验及结果分析

目的:本研究通过药敏实验对创伤弧菌进行抗菌药物敏感性分析,探讨在创伤弧菌感染中如何选择抗菌药物,提高临床治疗的效果.方法:利用ATCC标准的创伤弧菌菌株,进行了临床常用抗生素单个药敏实验和不同类型抗生素间的2种药物联合药敏实验.结果:创伤弧菌对抗革兰氏阴性菌的抗菌药物比较敏感,这些抗菌药物归属为(1)四环素类;(2)第三代头孢类;(3)第三、四代氟喹诺酮类;(4)新一代氧基糖甙类.四环素和头孢噻肟之间、氨苄西林和左旋氧氟沙星之间、复方新诺明和四环素之间等存在明显的协同关系,左旋氧氟沙星与头孢噻肟及四环素,头孢噻肟与庆大霉素及红霉素均不存在协同作用.结论:临床上主要应用抗菌药物对创伤弧菌感染进行对症治疗,本研究为临床中抗菌药物治疗创伤弧菌感染提供了实验依据,可作为临床用药和提高临床治疗效果的重要参考.     

大连地区海生物中致病性弧菌及气单胞菌分布的研究

本文首次报道了大连地区海产品中致病性弧菌及气单胞菌生态分布的研究结果。从152份海产品中共检出88株致病性弧菌及气单胞菌,检出率为57.9％.在检查欧氏六线鱼、太平洋鲱鱼、蓝色马鲛;蝼蛄虾;紫贻贝、杂色蛤仔和毛蚶共7种海产品中,检出9种致病性弧菌和1种气单胞菌,即溶藻弧菌、副溶血性弧菌、麦氏弧菌、少女鱼弧菌、弗尼斯弧菌、拟态弧菌、河弧菌、非01群霍乱弧菌和创伤弧菌。鱼虾类中菌株的检出率明显高于贝类。检出菌株中溶藻弧菌所占比例最高(54.5％),其次为副溶血性弧菌(27.3％)。首次从海生物中分离出嗜水气单胞菌、少女鱼弧菌等致病原因菌株.     

恩诺沙星对4种水产致病弧菌的抑杀菌效应

采用二倍稀释法测定恩诺沙星对溶藻弧菌、最小弧菌、哈维氏弧菌、创伤弧菌4种12株菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),结果表明:恩诺沙星对12株弧菌的MIC为0.1?0.8 mg/L,MBC为0.4?3.2 mg/L。在此基础上,从每种菌中选取一株对恩诺沙星较为敏感的菌株,研究恩诺沙星不同药物浓度(1 MIC、2 MIC、4 MIC)对4种弧菌的杀菌动力学和抗菌后效应(PAE),结果显示:各浓度恩诺沙星对溶藻弧菌X040625-R菌株均有较强的杀菌作用,而对哈维氏弧菌M071202-H、创伤弧菌Q050723-C、最小弧菌H010911-Z等3菌株却表现出了低浓度抑菌、高浓度缓慢杀菌的作用。恩诺沙星的抗菌后效应PAE与药物浓度及细菌与药物的接触时间成正相关,恩诺沙星对溶藻弧菌X040625-R的PAE最长,对哈维氏弧菌M071202-H的PAE最短。     

淡水养殖鱼类暴发性传染病致病细菌的研究

于1990—1991年的5—10月间,从湖北、湖南、河南和广东4省的9种淡水养殖鱼类的发病鱼睥、肝、肾中分离的93株细菌为材料,经100多个表型特征和细菌DNA中G+Cmol％遗传型特性测定等研究结果,可归属为产气单胞菌属中的运动性嗜中温产气单胞菌和弧菌属中的河弧菌。按照国际细菌命名法规优先律应为点状产气单胞菌[Aeromonas punctata(Zimmermann)nom.rev];而河弧菌与以前所描述的生物变种Ⅰ、Ⅱ型不同:在43℃能生长;发酵水杨苷产酸;利用D-葡糖酸钙、腐胺;不能利用戊二酸,D-葡糖醛酸;从D-葡萄糖产气;DNA中的G+Cmol％为47(Tm),鉴定为河弧菌生物变种Ⅲ(新生物变种Vibrio fluvialis Biovar Ⅲ.Biovar nov.)。     

2012年上海地区副溶血性弧菌血清分型和毒力基因携带状况研究

为了解2012年上海地区副溶血性弧菌人源株和食源株的优势血清型及其毒力基因携带状况,本研究收集了2012年从上海市15个区(县)腹泻患者和食品监测中分离的副溶血性弧菌株,进行血清分型,并采用聚合酶链反应(PCR)检测tdh和trh基因。结果显示,854株副溶血性弧菌中,88.1%为血清可分型,89.8%为产毒株。O3∶K6、O4∶K8、O1∶K25、O4∶K68、O4∶K9、O1∶K36、O3∶K29为上海地区可分型人源株的优势血清型(93.8%),其中O3∶K6最多,达56.2%。副溶血性弧菌全部分离株的月份分布显示出聚集趋势,7~8月为高峰期。O4∶K9和O1∶K36血清型菌株的月份分布与其他优势血清型菌株不同,未表现出明显聚集趋势。食源株无明显优势血清型,且与人源株分布不同。人源株产毒株构成(95.6%)高于食源株(5.5%)。人源株优势血清型产毒株构成(99.9%)高于非优势血清型(71.1%)。血清可分型人源株的tdh携带率(97.5%)高于不可分型人源株(67.6%),血清可分型人源株的trh携带率(0.8%)低于不可分型人源株(42.6%)。结果提示,副溶血性弧菌血清型分布与历史数据相比变化较大,血清型与毒力基因携带呈一定程度关联,且人源株与食源株在血清型和毒力基因携带上具有分离现象。因此,在副溶血性弧菌的监测与检测中应充分考虑血清分型和毒力基因的重要性。     

响应面分析法优化创伤弧菌生长条件

通过单因素分析,确定最适于创伤弧菌FJ04生长的p H、温度和盐度。在此基础上,综合考虑3个因素对创伤弧菌FJ04生长的影响,用Design-Expert软件进行响应面分析,优化FJ04的培养条件得到了菌体生长模型以及取得模型最优值时各因素的水平。结果表明:创伤弧菌FJ04的最优培养条件p H 8.29、温度32℃、盐度2.38%(质量分数);创伤弧菌生长过程中,温度和p H对创伤弧菌FJ04生长的交互作用显著,p H和盐度、温度和盐度对创伤弧菌FJ04生长的交互作用不显著,偏差度和准确度均在可接受范围内,模型是可靠的。     

低剂量创伤弧菌感染小鼠心肌与骨骼肌电镜超微结构观察比较

目的:观察低剂量创伤弧菌感染小鼠心肌、骨骼肌的超微结构变化,比较创伤弧菌引起的特征性病变下肢水肿骨骼肌病变与心脏病变出现的次序,探讨下肢水肿是否存在与心肌病变有关.方法:16只6～8周ICR(清洁级)小鼠.实验组12只腹腔注射＜LD50(1.34×107个/ml)的菌量(4.45×105个/ml)0.2ml,4只注射生理盐水0.2 ml作为菌液对照.分别取1 h、3 h、6 h、12 h小鼠心肌、后肢骨骼肌肌肉组织0.1cm×0.1 cm×0.1 cm置电镜固定液,超薄切片观察超微结构.结果:引起的动物模型实验组小鼠主要的实质性病变在心肌.实验组3 h就发现肌原纤维间隙扩大,肌丝断裂,肌膜下水肿较明显.6 h肌丝排列紊乱,疏松,局灶性肌丝断裂溶解.12 h核固缩水肿,间隙水肿,肌丝断裂,线粒体肿胀.而骨骼肌肌肉超微结构变化不明显,3 h、6 h和12 h实验组未死亡小鼠只表现肌组织间质变化,肌浆网扩张,组织间质水肿,胶原纤维排列稀疏、溶解.结论:本实验心肌、骨骼肌水肿与临床下肢水肿的症状相吻合,并提示心肌的实质病变明显早于骨骼肌,比较而言创伤弧菌所致病变是以重要脏器心脏的损伤为主,早期创伤弧菌对骨骼肌肌肉的实质性损伤并不明显.由创伤弧菌所致的原发性败血症表现双下肢出血性水肿可能不是创伤弧菌初始病变.     

环介导等温扩增联合横向流动试纸条快速检测创伤弧菌检测方法的建立

环介导等温扩增技术(LAMP)与横向流动试纸条(LFD)检测联合应用,建立了一种新的快速、便捷的创伤弧菌检测方法。针对创伤弧菌的外膜蛋白TolC基因设计6条特异性引物和1条异硫氰酸荧光素(FITC)标记的探针。生物素标记的LAMP扩增产物能够特异性地与FITC标记的探针杂交,杂交产物经LFD检测。优化后的扩增温度和时间为63℃反应35 min,加上细菌基因组DNA提取步骤,完成检测仅需要80 min。LAMP-LFD方法可特异性地检出创伤弧菌,对哈维氏弧菌等9种水产品常见病原菌的检测均呈阴性;对纯细菌培养物的检测灵敏度为3.7×102 CFU/mL或7.4 CFU/反应,是利用外引物建立的常规PCR检测的100倍。结果表明,该方法能够准确、快速、灵敏地检出创伤弧菌,可应用于创伤弧菌污染的水产品的检测。     

慢性肝病患者并发创伤弧菌脓毒症研究现状

创伤弧菌脓毒症,是由创伤弧菌引起的致死性疾病,主要见于慢性肝病患者。创伤弧菌是一种嗜盐的革兰阴性(G-)条件致病菌,自然存活于热带及亚热带海水中,可寄生在滤食海生动物体内(如牡蛎、蚌、蟹等)[1]。我国在浙江、江苏、广东、山东、广西等沿海地区贝壳类海产品中都曾检测到创     

副溶血弧菌Ⅲ型分泌系统vscG基因缺失株的构建及其生物学特性

【背景】副溶血弧菌(Vibrio parahaemolyticus)是一种重要的食源性病原微生物,Ⅲ型分泌系统(TypeⅢSecretionSystem,T3SS)在致病过程中发挥重要作用。vscG基因编码的Vsc G蛋白是Ⅲ型分泌系统的伴侣蛋白。vscG基因在副溶血弧菌中的生物学功能尚未确定。【目的】构建副溶血弧菌vscG基因的缺失株和回补株,研究vscG基因对副溶血弧菌生物学特性的影响。【方法】在副溶血弧菌POR-1菌株基础上利用同源重组法构建vscG基因的缺失株ΔvscG和回补株CΔvscG,分析比较POR-1、ΔvscG和CΔvscG在生长性能、生物被膜形成能力、运动性、溶血活性、细胞黏附和细胞毒性等方面存在的差异。【结果】与POR-1菌株相比,缺失株ΔvscG和回补株CΔvscG的生长特性和溶血活性无明显差异,缺失株ΔvscG的生物被膜形成能力下降,运动性较POR-1菌株和回补株CΔvscG增强。细胞感染试验显示,vscG基因的缺失明显降低了副溶血弧菌对HeLa细胞的黏附和毒性作用。【结论】vscG基因影响副溶血弧菌生物被膜的形成和运动性,在该菌黏附细胞和发挥细胞毒性过程中具有重要作用,为进一步探讨副溶血弧菌T3SS的致病机理奠定基础。     

Tuning of electronic properties of one-dimensional cyano-bridged Cu-Ni, Cu-Pd, and Cu-Pt bimetallic assemblies by stereochemistry of ligands

Preparations, XPS and electronic spectroscopy, and magnetism of seven new one-dimensional cyano-bridged coordination polymers, chiral [Cu(RR-chxn)₂][Pd(CN)₄] · 2H₂O (1), [Cu(trans-chxn)₂][M(CN)₄] · 2H₂O (2, 4, and 6 for M = Pd, Ni, and Pt), and [Cu(cis-chxn)₂][M(CN)₄] · 2H₂O (3, 5, and 7 for M = Pd, Ni, and Pt) (RR-chxn = cyclohexane-(1R,2R)-diamine, trans-chxn = racemic trans-cyclohexane-(1,2)-diamine, and cis-chxn = racemic cis-cyclohexane-(1,2)-diamine) have been reported in view of tuning of their electronic properties by stereochemistry of chxn ligands and metal-substitution. Comparison of Cu 2p_1/2 and 2p_3/2 peaks of XPS and broad d-d bands around 18 000 cm⁻¹ of electronic spectra are described systematically for 1-7. Variable-temperature magnetic measurement shows that complexes 1-7 indicate weak antiferromagnetic interactions via cyano-bridges. Because of semi-coordination coupled with pseudo Jahn-Teller elongation and electrostatic interaction for 1, the axial Cu-N coordination bond distances of 2.330(7) and 3.092(8) Å are considerably longer than those of equatorial ones in the range from 2.016(6) to 2.030(6) Å. The former bond distances of 1 are intermediate values among the related Ni (2.324(6) and 3.120(8) Å) and Pt (2.34(1) and 3.09(1) Å) complexes.     

Land,livestock, and livelihoods: Changing dynamics of gender,caste, and ethnicity in a Nepalese Village

Over the past 10 years, Ghusel VDC, Lalitpur District has moved from primarily subsistence agriculture into the wider cash economy aided by the Small Farmers' Development Program (SFDP), which provides credit to farmers mainly for the purchase of buffalo for milk production, and by the National Dairy Corporation, which supports local dairy cooperatives. Analysis reveals that buffalo-keeping and milk sales are increasing the well-being of many households, while at the same time creating new inequalities in gender roles and responsibilities, greater inequities between Brahmin and Tamang residents in Ghusel, and placing pressures on the ecosystem for increased supplies of fodder and fuelwood. Evidence suggests that there is critical, need for attention to the social, and particularly gender-based, implications of maintaining livestock for milk sales and to the ecological underpinnings of this livelihood system.     

Structure and function of S-adenosylhomocysteine hydrolase

In mammals, S-adenosylhomocysteine hydrolase (AdoHcyase) is the only known enzyme to catalyze the breakdown of S-adenosylhomocysteine (AdoHcy) to homocysteine and adenosine. AdoHcy is the product of all adenosylmethionine (AdoMet)-dependent biological transmethylations. These reactions have a wide range of products, and are common in all facets of biometabolism. As a product inhibitor, elevated levels of AdoHcy suppress AdoMet-dependent transmethylations. Thus, AdoHcyase is a regulator of biological transmethylation in general. The three-dimensional structure of AdoHcyase complexed with reduced nicotinamide adenine dinucleotide phosphate (NADH) and the inhibitor (1′R, 2′S, 3′R)-9-(2′,3′-dihyroxycyclopenten-1-yl)adenine (DHCeA) was solved by a combination of the crystallographic direct methods program, SnB, to determine the selenium atom substructure and by treating the multiwavelength anomalous diffraction data as a special case of multiple isomorphous replacement. The enzyme architecture resembles that observed for NAD-dependent dehydrogenases, with the catalytic domain and the cofactor binding domain each containing a modified Rossmann fold. The two domains form a deep active site cleft containing the cofactor and bound inhibitor molecule. A comparison of the inhibitor complex of the human enzyme and the structure of the rat enzyme, solved without inhibitor, suggests that a 17° rigid body movement of the catalytic domain occurs upon inhibitor/substrate binding.     

Chiral Separation of G‐type Chemical Warfare Nerve Agents via Analytical Supercritical Fluid Chromatography

Chemical warfare nerve agents (CWNAs) are extremely toxic organophosphorus compounds that contain a chiral phosphorus center. Undirected synthesis of G‐type CWNAs produces stereoisomers of tabun, sarin, soman, and cyclosarin (GA, GB, GD, and GF, respectively). Analytical‐scale methods were developed using a supercritical fluid chromatography (SFC) system in tandem with a mass spectrometer for the separation, quantitation, and isolation of individual stereoisomers of GA, GB, GD, and GF. Screening various chiral stationary phases (CSPs) for the capacity to provide full baseline separation of the CWNAs revealed that a Regis WhelkO1 (SS) column was capable of separating the enantiomers of GA, GB, and GF, with elution of the P(+) enantiomer preceding elution of the corresponding P(–) enantiomer; two WhelkO1 (SS) columns had to be connected in series to achieve complete baseline resolution. The four diastereomers of GD were also resolved using two tandem WhelkO1 (SS) columns, with complete baseline separation of the two P(+) epimers. A single WhelkO1 (RR) column with inverse stereochemistry resulted in baseline separation of the GD P(–) epimers. The analytical methods described can be scaled to allow isolation of individual stereoisomers to assist in screening and development of countermeasures to organophosphorus nerve agents. Chirality 26:817–824, 2014. © 2014 The Authors. Chirality published by John Wiley Periodicals, Inc.     

Influence of Fluoride on Rat Kidney Antioxidant System: Effects of Methionine and Vitamin E

The aim of the study has been to determine and compare the influence upon the kidney antioxidative system, exercised by administration of vitamin E, and vitamin E in combination with methionine, under conditions of oxidative stress induced by sodium fluoride. The experiment was carried out on Wistar FL rats (adult males) that, for 35 days, were administered water, NaF, NaF with vitamin E, or vitamin E with methionine (doses: 10 mg NaF/kg of body mass/24 h, 3 mg vitamin E per 10 μl per rat for 24 h, 2 mg methionine per rat for 24 h). The influence of administered sodium fluoride and antioxidants upon the antioxidative system in kidney was examined by analyzing the concentration of malondialdehyde (MDA) and the activity of the most important antioxidative enzymes (SOD, total and both its isoenzymes, GPX, GST, GR, and CAT). The studies carried out confirmed the disadvantageous effect of the administered dose of NaF upon the antixodiative system in rats (increase in the concentration MDA, decrease activity of all antioxidative enzymes). The administration of vitamin E increased the activity of studied enzymes with the exception of glutathione reductase GR; it also reduced the procesess of lipid peroxidation. It has been found that combined doses of vitamin E and methionine were most effective in inhibiting lipid peroxidation processes. The results confirmed the antioxidative properties of methionine.     

Reorientation of Microfibrils and Microtubules at the Outer Epidermal Wall of Maize Coleoptiles During Auxin-Mediated Growth

Auxin-mediated elongation growth of isolated subapical coleoptile segments of maize (Zea mays L.) is controlled by the extensibility of the outer cell wall of the outer epidermis (Kutschera et al., 1987). Here we investigate the hypothesis that auxin controls the extensibility of this wall by changing the orientation of newly deposited microfibrils through a corresponding change in the orientation of cortical microtubules. On the basis of electron micrographs it is shown that cessation of growth after removal of the endogenous source of auxin is correlated with a relative increase of longitudinally orientated microfibrils and microtubules at the inner wall surface. Conversely, reinduction of growth by exogenous auxin is correlated with a relative increase of transversely orientated microfibrils and microtubules at the inner wall surface. These changes can be detected 30–60 min after the removal and addition of auxin, respectively. The functional significance of directional changes of newly desposited wall microfibrils for the control of elongation growth is discussed.     

Gamma-glutamylcysteine protects ergothioneine-deficient Mycobacterium tuberculosis mutants against oxidative and nitrosative stress

Mycobacterium tuberculosis (M.tb.), the causative agent of tuberculosis (TB), cannot synthesize GSH, but synthesizes two major low molecular weight thiols namely mycothiol (MSH) and ergothioneine (ERG). Gamma-glutamylcysteine (GGC), an intermediate in GSH synthesis, has been implicated in the protection of lactic acid bacteria from oxidative stress in the absence of GSH. In mycobacteria, GGC is an intermediate in ERG biosynthesis, and its formation is catalysed by EgtA (GshA). GGC is subsequently used by EgtB in the formation of hercynine-sulphoxide-GGC. In this study, M.tb. mutants harbouring unmarked, in-frame deletions in each of the fives genes involved in ERG biosynthesis (egtA, egtB, egtC, egtD and egtE) or a marked deletion of the mshA gene (required for MSH biosynthesis) were generated. Liquid chromatography tandem mass spectrometry analyses (LC-MS) revealed that the production of GGC was elevated in the MSH-deficient and the ERG-deficient mutants. The ERG-deficient ΔegtB mutant which accumulated GGC was more resistant to oxidative and nitrosative stress than the ERG-deficient, GGC-deficient ΔegtA mutant. This implicates GGC in the detoxification of reactive oxygen and nitrogen species in M.tb.     

In vitro studies of the rabbit immune system. IV. Differential mitogen responses of isolated T and B cells.

The mitogenic responses of separated rabbit lymphocyte populations functionally analogous to mouse T and B cells have been tested in vitro. Purified T cells were prepared by passage over nylon wool (NW) and purified B cells prepared by treatment with antithymocyte serum and complement (ATS + C). ATS + C kills 70% of peripheral blood lymphocytes (PBL's) and 50% of the spleen cells while passage over NW yields 40% of the applied PBL's and 5–23% of the applied spleen cells. NW-purified T cells from the spleen or PBL's respond fully to concanavalin A (Con A) but have a reduced response to phytohemaglutinin (PHA) and little or no response to goat anti-rabbit immunoglobulin (anti-Ig). PBL's that survive ATS + C (B cells) are stimulated by anti-Ig but not by Con A or PHA. B cells purified from spleen do not respond to Con A or PHA but will respond to anti-Ig under appropriate conditions. A full spleen B-cell response to anti-Ig required removal of Ig produced by the cultures that blocked anti-Ig stimulation. It is concluded that, for rabbit lymphocytes, Con A and PHA are primarily T-cell mitogens and that anti-Ig is primarily a B-cell mitogen. However, the mitogen response of unfractionated PBL or spleen cell populations indicates an overlap in reactivity. This could be due to cells sharing T and B properties, alteration of cell populations by the fractionation procedures used, or recruitment of one population in the presence of a mitogenic response of the other population.     

八种脑-肠肽侧脑室内注射对大鼠基础胃酸分泌的影响

用乌拉坦麻醉大鼠作急性实验,采用连续灌流胃并收集流出液的方法,观察向侧脑室内注射微量脑-肠肽对大鼠基础胃酸分泌的影响。实验结果如下:(1)雨蛙肽、八肽胆囊收缩素、促甲状腺素释放激素及四肽胃泌素均使总酸排出量增加;(2)生长抑素、胰多肽、P 物质、胰高血糖素则使总酸排出量减少;(3)上述肽类用侧脑室注射的剂量作肌肉注射,除四肽胃泌素也产生明显的刺激胃酸分泌作用外,对胃酸分泌均无明显影响。以上结果提示,脑内的一些肽类可能以神经递质或调制物的方式,参与中枢对胃酸分泌的调节。     

A chromium-tolerant plant growing in Cr-contaminated land

In this paper, a kind of Typhaceae plant species, Typha angustifolia L, with a high tolerance to Cr is described. Experiments were carried out to examine its ability to tolerant Cr and its physiological response. The results showed that there was no difference in growth, plant height, and biomass response to external Cr (VI) between the plants exposed to 100 microM Cr (VI) and control (0 microM), while increasing Cr levels to 200-800 microM induced a significant decrease in plant height and biomass, but no significant injury was detected, even for the plants exposed to 800 microM Cr. Chromium induced significant increases in superoxide dismutase (SOD) and peroxidase (POD) activities. Meanwhile, a significantly positive correlation was found between Cr and Mn or Cu in leaves and roots, respectively. The Cr tolerance of the plant appeared to be associated with the enhancement of SOD and POD activities and the improvement in uptake and translocation of the essential microelements.