Thoracic spine infection caused by Pseudomonas fluorescens:A case report and review of literature

BACKGROUND The clinical incidence of spinal infection is gradually increasing,and its onset is insidious,easily leading to missed diagnosis and misdiagnosis,which may lead to serious complications such as nervous system dysfunction,spinal instability and/or deformity,and cause a huge burden on society and families.Early identification of the causative agent and precision medicine will greatly reduce the suffering of patients.At present,the main pathogenic bacteria that cause spinal infection are Staphylococcus aureus,Streptococcus,Pneumococcus,Escherichia coli,and Klebsiella.There are no reports of spinal infection caused by Pseudomonas fluorescens.CASE SUMMARY We report a 32-year-old female patient with spinal infection.She presented with flank pain,initially thought to be bone metastases or bone tuberculosis,and had a family background of tumors.Her clinical features and changes in imaging and laboratory tests led to the suspicion of thoracic spine infection.Histopathology of the lesion showed inflammation,tissue culture of the lesion was negative several times,and the possible pathogen-Pseudomonas fluorescens was found after gene sequencing of the lesion.The patient recovered completely after a full course of antibiotic treatment.CONCLUSION This report increases the range of pathogens involved in spinal infections,highlights the unique advantages of gene sequencing technology in difficult-todiagnose diseases,and validates conservative treatment with a full course of antibiotics for spinal infections without complications.

Pseudomonas fluorescens-like bacteria from the stomach:A microbiological and molecular study

AIM:To characterize oxidase-and urease-producing bacterial isolates,grown aerobically,that originated from antral biopsies of patients suffering from acid peptic diseases.METHODS:A total of 258 antral biopsy specimens were subjected to isolation of bacteria followed by tests for oxidase and urease production,acid tolerance and aerobic growth.The selected isolates were further characterized by molecular techniques viz.amplifications for 16S rRNA using universal eubacterial and HSP60 gene specific primers.The amplicons were subjected to restriction analysis and partial sequencing.A phylogenetic tree was generated using unweighted pair group method with arithmetic mean(UPGMA) from evolutionary distance computed with bootstrap test of phylogeny.Assessment of acidity tolerance of bacteria isolated from antrum was performed using hydrochloric acid from 10-7 mol/L to 10-1 mol/L.RESULTS:Of the 258 antral biopsy specimens collected from patients,179(69.4%) were positive for urease production by rapid urease test and 31%(80/258) yielded typical Helicobacter pylori(H.pylori) after 5-7 d of incubation under a microaerophilic environment.A total of 240(93%) antral biopsies yielded homogeneous semi-translucent and small colonies after overnight incubation.The partial 16S rRNA sequences revealed that the isolates had 99% similarity with Pseudomonas species.A phylogenetic tree on the basis of 16S rRNA sequences denoted that JQ927226 and JQ927227 were likely to be related to Pseudomonas fluorescens(P.fluorescens).On the basis ofHSP60 sequences applied to the UPGMA phylogenetic tree,it was observed that isolated strains in an aerobic environment were likely to be P.fluorescens,and HSP60 sequences had more discriminatory potential rather than 16S rRNA sequences.Interestingly,this bacterium was acid tolerant for hours at low pH.Further,a total of 250(96.9%) genomic DNA samples of 258 biopsy specimens and DNA from 240 bacterial isolates were positive for the 613 bp amplicons by targeting P.fluorescens-specific conserved putative outer membrane protein gene sequences.CONCLUSION:This study indicates that bacterial isolates from antral biopsies grown aerobically were P.fluorescens,and thus acid-tolerant bacteria other than H.pylori can also colonize the stomach and may be implicated in pathogenesis/protection.

Identification and Characterization of Genes Responsible for Drought Tolerance in Rice Mediated by Pseudomonas fluorescens

Drought is one of the major abiotic stresses which adversely affect crop plants limiting growth and yield potential.Structural and functional characterization of drought stress-induced genes has contributed to a better understanding of how plants respond and adapt to the drought stress.In the present study,differential display technique was employed to study the gene expression of rice plants at the reproductive stage that were subjected to drought stress by withholding water,Pseudomonas fluorescens strain(Pf1) treated plants subjected for drought stress by withholding water and control(well-watered).Differentially expressed c DNAs of six genes(COX1,PKDP,b ZIP1,AP2-EREBP,Hsp20 and COC1) were identified,cloned and sequenced.Real-time q PCR analysis showed that all the six genes were upregulated in drought-stressed plants treated with Pf1.This revealed that the remarkable influence of Pf1 colonization leads to drought tolerance at the reproductive stage.These results showed that high levels of gene expression in plants lacking adequate water can be remarkably influenced by Pf1 colonization,which might be a key element for induced systemic tolerance by microbes.

Production and Optimization of <i>Pseudomonas fluorescens</i>Biomass and Metabolites for Biocontrol of Strawberry Grey Mould

Pseudomonas species have been widely studied as biological agents (BCAs) and it is alternative to the application of chemical fungicides. Our objective was to optimize nutritional and environmental conditions of the isolated Pseudomonas fluorescens fp-5 for biomass and metabolites production and to evaluate itsagainst the grey mould disease caused by Botrytis cinerea on strawberry plants under field conditions. Pseudomonas fluorescens, showed antagonistic properties, in vitro, against thepathogen Botrytiscinerea. Effect of the separated secondary metabolites on the fungal growth by broth dilution technique and antifungal activity by agar well diffusion technique was studied. Response surface methodology was used to investigate the effects of four fermentation parameters (pH, incubation time, carbon and nitrogen concentrations) on biomass and bioactivemetabolites [antibiotic phenazin and siderophore] production. Glycerol was found to be the best carbon source for improved biomass and metabolites production. Meanwhile, peptone and yeast extract were found to be the best nitrogen source. Analysis of each formulation revealed that glycerol oil at 0.01% the best oil used for protect P. fluorescens for 3 months Under natural condition, P. fluorescens formulation was effective in reducing B. cinerea disease in strawberry leaves and fruits. Pre-harvest treatment protected fruits from Botrytis post-harvest disease in comparing of fungicide. In addition, the obtained results showed that bacterial treatment significantly increased thegrowth parameters as well as dry weights and yield.

Bacteremia or pseudobacteremia?Review of pseudomonas fluorescens infections

INTRODUCTION P.fluorescens is an aerobic,Gram-negative bacillus related to Pseudomonas aeruginosa.Like other species of Pseudomonas,the organism is widespread in nature and is found in water,moist soil,and vegetation.Due to its low virulence,P.fluorescens is an infrequent cause of infections except for catheter-related bloodstream infections in cancer patients.From August 1,2003 to May 31,2016,Hyogo Emergency Medical Center and the Kobe Red Cross Hospital treated three cases of bloodstream infection

糖蜜废水培养微生物絮凝剂产生菌Pseudomonas fluorescensC-2及优化

利用糖蜜废水驯化、培养微生物絮凝剂产生菌Pseudomonas fluorescensC-2,通过单因素试验和正交试验设计优化得到该菌株产絮凝剂的最佳培养条件：培养基C∶N∶P为100∶5∶1,培养时间为48 h,培养基初始pH值8.0,糖蜜废水COD浓度8 000 mg·L^-1,培养温度为30℃,摇床转速为150 r·min^-1.在此培养条件下产生的絮凝剂对高岭土悬浊液絮凝率达94.75%,且对多种废水有较好的净化效果,对废水中色度和浊度的去除率均在80%以上,对COD的去除率为53.66%-85.33%,说明利用糖蜜废水培养絮凝剂产生菌C-2是完全可行的.

Improvement of Bio-Efficacy to Reduce Bacterial Wilt Complex Disease in Tomatoes through Bacillus amyloliquefaciens and Pseudomonas fluorescens under Field Conditions

Bacterial wilt complex disease of tomato(Solanum lycopersicum L.)was incited jointly by bacterial wilt pathogen Ralstonia solanacearum and Meloidogyne incognita worldwide.Bio-efficacy of bacterial antagonists i.e.B.amyloliquefaciens DSBA-11 and P.fluorescens DTPF-3 was studied against the wilt disease complex in tomato at National Phytotran facility Indian Agricultural Research Institute(IARI),New Delhi,at 26±2°C.Minimum wilt disease incidence(26.00%)with the highest bio-control efficacy(64.15%),less juvenile population(19.33 J2/g of soil)of M.incognita was recorded in the combined application of DTPF-3+DSBA-11 after 30 d of inoculation under glasshouse conditions.In a field study,minimum bacterial wilt disease incidences 19.0%and 20.4%were recorded in the bleaching powder treatment followed by mixed application of DSBA-11+DTPF-3,19.6%and 21.2%wilt incidence in 2014 and 2015 respectively.However,a reduction of root-knot gall index was recorded a maximum of 59.76%and 69.62%in DSBA-11+DTPF-3 treated plants followed by 54.88%and 60.13%over control in DTPF-3 treatment in 2014 and 2015 respectively.The yield of tomato fruit was increased over control by 17.48%and 16.97%in 2014 and 2015 respectively under field conditions.A combination of P.fluorescens DTPF-3+B.amyloliquefaciens DSBA-11 suppressed bacterial wilt and root-knot diseases and also increased the yield of the tomato fruit significantly(p<0.05)under field conditions.

P. fluorescens and B. megaterium Effect on the Lifespan of Mutant-Type dpy-11 and Wild-Type of C. elegans

The research question being studied in this paper is how do different types of bacteria as food (Pseudomonas fluorescens and Bacillus megaterium) affect the lifespan of Caenorhabditis elegans in dpy-11 mutant-type and wild-type? P. fluorescens and B. megaterium will be the two pathogens that will be tested on two different types of C. elegans: mutant-type dpy-11 and wild-type. From the analysis of primary articles studying these pathogens, it can be concluded that P. fluorescens and B. megaterium are decent contenders for allowing C. elegans to grow and possibly extend the lifespan of it. P. fluorescens will allow the lifespan of the two types of nematodes to be longer. Additionally, the mu-tant-type dpy-11 of C. elegans will have a much longer lifespan, even double, compared to that of the wild-type. The results showed P. fluorescens had a longer lifespan than B. megaterium but not as long as C. elegans’ main food source, E. coli. C. elegans mutant dpy-11 had a longer lifespan than the wild-type. Furthermore, there were no C. elegans present in the B. megaterium wild-type plates.

Penetration and Post-infection Development of Meloidogyne Incognita on Tobacco as Treated by Rhizobacteria Pseudomonas Fluorescens Strain P-72-10

The bio-control potential of rhizospere bacteria Pseudomonas fluorescens against plant-parasite nematode had been demonstrated. P. fluorescens had shown the effect to enhance tobacco resistance to root-knot nematode Meloidogyne incognita. Inoculation with P. fluorescens in tobacco could lead to significant reductions in the number of juveniles that penetrated tobacco root and further life stage development of the juveniles. The number of juveniles penetrated into tobacco root in treatment with P. fluorescens is significantly different from CK at 2DAI,6DAI,8DAI and 10 DAI. Significant reduction and delayed development of juveniles that penetrated into tobacco root and treated were observed in treatment at14 DAI,21DAI,28 DAI and 35 DAI. In addition,P. fluorescens treatment leads to a significant reduction in the number of eggs per egg-mass at 35 DAI. The results show P. fluorescens induced a continuously suppression on root-knot nematode M. incognita throughout their entire early infection phase of root penetration,subsequent life stage development and reproduction.

Biosorption of cesium(Ⅰ) from aqueous solution by a novel exopolymers secreted from Pseudomonas fluorescens C-2: Equilibrium and kinetic studies

The biosorption characteristics of Cs（I） ions from aqueous solution using exopolymers （PFC02） produced from Pseudomonas fluorescens C-2 were investigated as a function of pH, biosorbent dosage, contact time and initial concentration. pH played a major role in the adsorption process, and the optimum pH for the removal of Cs（I） was 8.0. Langmuir, Freundlich and Dubinin-Radushkevich （D-R） models were applied to describe the biosorption isotherm of the Cs（I） ions by PFC02. The Lagergren first-order, pseudo second-order kinetic and intraparticle diffusion models were used to test the kinetic data. Langmuir model and D-R model fitted the equilibrium data better than the Freundlich isotherm. The monolayer adsorption capacities of PFC02 as obtained from Langmuir isotherm at 25°C was found to be 32.63 mg/g. From the D-R isotherm model, the mean free energy was calculated as 26.73 kJ/mol, indicating that the biosorption of cesium was chemisorption. The biosorption process was rapid, and the kinetic rates were best fitted to the pseudo second-order model, which indicated the biosorption process operated through chemisorption mechanism. FT-IR analysis of PFC02 showed the possible functional groups responsible for cesium adsorption were hydroxyl, carboxyl, carbonyl and sulphonate groups. SEM analysis showed the porous structure of the material while EDX analysis confirmed the adsorption of Cs（I） on PFC02. Cesium adsorbed onto the PFC02 could be desorbed efficiently using 1 mol/L HNO 3 , and the enrichment factor was 50.0. Furthermore, PFC02 could be reused five times with only about 8.25% regeneration loss. The developed method was successfully utilized for the removal of Cs（I） ions from aqueous solution.

一株草鱼源荧光假单胞菌的分离鉴定及其耐药特性分析

为探究临沂市某草鱼养殖场发生病害的原因,从患病的草鱼(Ctenopharyngodon idellus)肝脏和肾脏混合组织中分离得到一株优势致病菌G-13,通过细菌形态学观察、革兰氏染色、生理生化特征、16S rRNA基因测序,以及系统发育分析等完成菌株鉴定,并测定其耐药特性。形态特征显示G-13是一种短杆状的革兰氏阴性菌,在LB固体平板上形成乳白色、圆形、整体呈隆起状、边缘较为整齐、表面光滑湿润的菌落;16S rRNA系统发育进化分析发现G-13与荧光假单胞菌(Pseudomonas fluorescens)聚在一支,序列同源性高达100%;生理生化试验显示葡萄糖、精氨酸双水解酶、鸟氨酸脱羧酶、赖氨酸脱羧酶、氧化酶、无盐和3%NaCl胰胨水等7项指标为阳性,与假单胞菌属特征相近,确定分离株G-13为P.fluorescens。药敏试验结果显示菌株G-13对丁胺卡那、多西环素、四环素、新霉素、环丙沙星、氧氟沙星等13种抗生素敏感,对氨苄西林、头孢拉定、克林霉素、呋喃唑酮等7种抗生素耐药。

纳米TiO_2对鸡肉源微生物的光催化抑菌效果研究

[目的]研究不同作用因素对纳米TiO_2的光催化活性影响,为纳米TiO_2光催化杀菌技术的开发与应用提供参考。[方法]以菌液初始浓度、光照强度和纳米TiO_2浓度为主要变量因素,研究不同因素条件下纳米TiO_2对鸡肉源微生物P.fluorescens和M.caseolyticus的光催化抑菌作用效果。[结果]纳米TiO_2浓度增加对P.fluorescens和M.caseolyticus的抑菌作用呈先升高后降低的趋势,最佳浓度值为0.4 g/L;增高光照强度,降低初始菌液浓度,纳米TiO_2的光催化抑菌效果增强;相同试验条件下,纳米TiO_2对P.fluorescens和M.caseolyticus的光催化抑菌作用一致,对M.caseolyticus的抑菌率始终低于P.fluorescens。[结论]M.caseolyticus对纳米TiO_2的光催化作用比P.fluorescens更加敏感。

荧光假单胞菌BIT-18磷脂酶B基因的克隆及生物信息学分析

利用PCR从Pseudomonas fluorescens BIT-18总DNA中成功扩增到编码Pf-PLB的全长基因,并进行测序.通过生物信息学分析,可知Pf-PLB基因全长1272 bp,编码423个氨基酸,理论分子量为45.8kDa,等电点为5.53,在N末端有一个包含23个氨基酸的信号肽.进化树及序列分析结果显示,pfplb是PLBs新基因家族的成员.采用模建软件Modeller进行人工建模,结果表明Pf-PLB为由14股Strand组成的β-桶状蛋白,本研究为Pf-PLB进一步高效表达及其脱胶机理研究奠定基础.

产2,4-二乙酰基间苯三酚的微生物研究进展

在土壤环境中,大多数2,4-二乙酰基间苯三酚(2,4-DAPG)是由荧光假单胞菌(Pseudomonas fluorescens)产生的。它在生物防治中具有重要作用。对近年来2,4-DAPG的生物合成及调控机理,2,4-DAPG在诱导系统抗性(ISR)的机制,2,4-DAPG的生物反应模式、生态效应,荧光假单胞菌农田生物防治应用实例等相关研究进行了综述。

来源于假单孢菌206植酸酶基因的克隆、表达及酶学性质研究

从浙江嘉兴人工鱼塘底泥样品筛选到产植酸酶活性较高的假单孢菌(Pseudomonas fluorescens)206。通过简并PCR和TAIL-PCR技术从Pseudomonas fluorescens206菌株基因组DNA中克隆得到一个新的编码组氨酸酸性植酸酶(HAP)的基因,命名为phyP。该基因ORF全长1 284 bp,编码了427个氨基酸和一个终止密码子,前24个氨基酸为信号肽。将phyP在大肠杆菌中表达,重组蛋白经硫酸铵分级沉淀和阴离子交换柱纯化后达到电泳纯。对其酶学性质分析表明:重组植酸酶PhyP最适pH为5.5,在pH3.5-7.5的条件下具有较好的稳定性;重组植酸酶PhyP最适温度为45℃,在25℃时也具有较高的酶活性,因此植酸酶PhyP在饲料行业,尤其是水产行业具有潜在的应用前景。

染色体荧光原位杂交技术研究小片段和复杂的染色体易位

染色体之间的平衡易位，当片段很小或涉及染色体数目多时，显微镜下容易漏诊，或难以准确判定断裂点。本文报告了用染色体荧光原位杂交（ＦＩＳＨ）的实验技术结合常规Ｇ显带、高分辨Ｇ显带技术诊断了２例难辨认的染色体易位。本研究表明，ＦＩＳＨ技术结合细胞遗传学核型分析，对于检测小片段难辨认或复杂的染色体易位是非常有帮助的。

全自动免疫组化和荧光原位杂交方法检测胃癌HER2状态

目的探讨胃癌中HER2基因状态分布及HER2基因扩增/HER2蛋白表达与临床病理特征间的关系。方法收集四川大学华西医院2009年1月~3月170例手术切除原发性胃或胃食管交界处腺癌石蜡组织标本,同时收集患者的临床和病理信息。所有病例行全自动免疫组化(IHC)和荧光原位杂交(FISH)检测HER2蛋白和基因的表达。结果 170例胃癌组织中,IHC检测蛋白表达为:100例(58.8%)0+,33例(19.4%)1+,14例(8.2%)2+,23例(13.6%)3+。FISH检测HER2基因扩增27例(15.9%),17号染色体多倍体者20例(70.4%)。其中4例(4%)IHC检测0+或1+,FISH检测阳性;有14例(37.8%)IHC检测2+或3+,FISH检测阴性。IHC与FISH结果间差异显著(P<0.01)。HER2过表达和基因扩增与患者的年龄、性别、病理分级、肿瘤部位及TNM分期间没有相关性(P>0.05),与分化程度、Lauren分级有相关性(P<0.01)。结论IHC与FISH检测胃癌HER2基因的阳性率分别为21.7%和15.9%。HER2基因扩增中17号染色体多倍体发生比例较高。分化程度越高,HER2阳性率越高,且主要集中在肠型。IHC可作为胃癌HER2检测的有效筛查手段,其不确定的病例可再行FISH检测确诊。

Rhizosphere Bacteria for Biocontrol of Bacterial Blight and Growth Promotion of Rice

Several bacterial strains were isolated from different rhizospheres. Among these, strain PDY7 exhibited strong antibacterial activity against the rice bacterial blight （BB） pathogen Xanthomonas oryzae pv. oryzae （Xoo） by the laboratory dual plate assays. The antibacterial property of the strain PDY7 was further investigated for the production of 2,4-diacetylphloroglucinol （DAPG）, which amplified a characteristic of 629-bp DNA fragment by PCR-based screening method using phlD primers. The application of phlD positive strains was carefully evaluated for disease control and growth promotion of rice plants under field conditions. The selected strain PDY7 suppressed the rice BB by 58.83% and 51.88% under glass house and field conditions, respectively. In addition, the strain PDY7 showed significant two-fold increase in root length （18.08 cm）, shoot length （29.81 cm）, and grain yield （96.07 g）. Strain PDY7 promoted the growth of rice plants by production of indole-3-acetic acid （IAA）, which was determined by high performance liquid chromatography （HPLC） analysis. Our findings suggest that PDY7 belongs to the P. fluorescens group and can serve as potential biocontrol of BB as well as biofertilizer agent for growth promotion of rice.

萘环荧光离子液体合成、表征及光谱性能研究

以1-氯甲基萘和N-烷基咪唑为原料,四氢呋喃为溶剂,合成了5个具有荧光的新型离子液体。采用红外光谱(FT-IR)、核磁共振氢谱(1H NMR)、碳谱(13C NMR)等测试技术对产物进行了结构表征。并对其紫外光谱、荧光光谱等光谱性质进行了初步的研究。实验结果表明咪唑离子液体1.2二甲基-4-甲基萘咪唑氯化盐较咪唑离子液体1-烷基-3-甲基萘咪唑氯化盐的荧光的激发和发射光谱的波长大,且荧光量子产率高。

头孢氨苄血药浓度的荧光法测定及药物动力学研究

本文研究了荧光法测定头孢氨苄的血药浓度及其药物动力学。实验结果表明,血清浓度在1～20μg/ml范围内,荧光强度与浓度有较好的线形关系,r=0.9975。回收率为94.7%±2.3%(n=5),CV=2.39%。浓度时间曲线(头孢氨苄0.5g,PO)显示为一室模型。平均药物动力学参数(n=8):Ka=1.7500(h^(-1)),K=0.7365(h^(-1)),T_(1/2)=0.9401(h),T_(max)=1.18(h),C_p=19.64(μg/ml),V_a/F=13568.2(ml)。