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1.

Background  

Pseudomonas aeruginosa is commonly associated with contact lens (CL) -related eye infections, for which bacterial adhesion and biofilm formation upon hydrogel CLs is a specific risk factor. Whilst P. aeruginosa has been widely used as a model organism for initial biofilm formation on CLs, in-vitro models that closely reproduce in-vivo conditions have rarely been presented.  相似文献   

2.
Dental unit water line (DUWL) contamination by opportunistic pathogens has its significance in nosocomial infection of patients, health care workers, and life-threatening infections to immunocompromized persons. Recently, the quorum sensing (QS) system of DUWL isolates has been found to affect their biofilm-forming ability, making it an attractive target for antimicrobial therapy. In this study, the effect of two quorum-sensing inhibitory compounds (patulin; PAT, penicillic acid; PA) and EDTA on planktonic growth, AI-2 signalling and in vitro biofilm formation of Pseudomonas aeruginosa, Achromobacter xylosoxidans and Achromobacter sp. was monitored. Vibrio harveyi BB170 bioassay and crystal violet staining methods were used to detect the AI-2 monitoring and biofilm formation in DUWL isolates, respectively. The V. harveyi BB170 bioassay failed to induce bioluminescence in A. xylosoxidans and Achromobacter sp., while P. aeruginosa showed AI-2 like activity suggesting the need of some pretreatments prior to bioassay. All strains were found to form biofilms within 72 h of incubation. The QSIs/EDTA combination have isolate-specific effects on biofilm formation and in some cases it stimulated biofilm formation as often as it was inhibited. However, detailed information about the anti-biofilm effect of these compounds is still lacking.  相似文献   

3.
The gene (vgb) encoding the hemoglobin (VtHb) of Vitreoscilla sp. was cloned into a broad-host-range vector and stably transformed into Pseudomonas putida, Pseudomonas aeruginosa, and Xanthomonas maltophilia. vgb was stably maintained and expressed in functional form in all three species. When growth of the P. aeruginosa and X. maltophilia transformants in Luria-Bertani medium was compared with that of each corresponding untransformed strain, the VtHb-producing strains reached slightly higher maximum viable cell numbers, had significantly increased viability after extebded times in culture, and, like E. coli that produces VtHb, had significantly lower respiration rates. The VtHb-producing strain of P. putida also reached a slightly higher maximum viable cell number than its corresponding untransformed strain, but was significantly less viable after extended times in culture and, unlike the case in E. coli, had a generally higher respiration rate than the untransformed strain. When growth was monitored by absorbance, the results were similar to those obtained with viable cell counts.  相似文献   

4.
This study was designed to evaluate the effects of the inorganic electrolytes present in human tear film on the viability and colonization of bacteria to hydrogels. Pseudomonas aeruginosa and Staphylococcus aureus were used in these experiments. A D-value test was performed to investigate any bacteriostatic effect by measuring the reduction of viable test microorganisms over time when exposed to the inorganic electrolyte solution. No D-value was calculable for S. aureus in electrolyte solution whereas a D-value of 8.1 h was obtained for P. aeruginosa in electrolyte solution. The D-value data indicate that staphylococci have a greater survivability potential in a hypertonic environment than do pseudomonads. Bacterial adhesion to high water, ionic hydrogels was studied using the Modified Robbins Device (MRD). The data for P. aeruginosa recovered from the lenses showed an 82% decrease in bacterial counts in electrolyte solution as compared to bacteria incubated in control solution. In contrast there were slight increases in S. aureus counts recovered from the lenses. Journal of Industrial Microbiology & Biotechnology (2000) 25, 17–19. Received 12 August 1999/ Accepted in revised form 05 January 2000  相似文献   

5.
PurposeAcanthamoeba keratitis (AK) is a painful and potentially blinding corneal infection caused by Acanthamoeba spp. In Madrid, environmental studies have demonstrated a high presence of these free-living amoebae in tap water. Since most of AK cases occur in contact lenses (CL) wearers with inadequate hygiene habits, the presence of Acanthamoeba in discarded CL has been studied and compared with other common etiological agents of keratitis, such as Pseudomonas aeruginosa and Staphylococcus aureus.MethodsOne hundred and seventy-seven healthy individuals from Madrid contributed their discarded CL and answered a questionnaire on hygiene habits. DNA was extracted from the CL solution and analyzed by real-time PCR for Acanthamoeba, Pseudomonas aeruginosa and Staphylococcus aureus. These CL and their solutions were also cultured on non-nutrient agar to isolate Acanthamoeba.ResultsAmong the 177 samples, Acanthamoeba DNA was detected in 87 (49.2%), P. aeruginosa DNA in 14 (7.9%) and S. aureus DNA in 19 (10.7%). Cultivable amoebae, however, were observed in only one sample (0.6%). This isolate was genotyped as T4. The habits reported by this CL owner included some recognized risk factors for AK, but in this study only the practice of “not cleaning the CL case” presented some statistical significant association with Acanthamoeba DNA presence. Detection of the investigated bacterial DNA did not demonstrate statistical significant association with the studied practices, but the presence of P. aeruginosa revealed a possible inhibition of Acanthamoeba in these samples.ConclusionsThe PCR results suggest a high presence of Acanthamoeba spp. in healthy CL wearers from Madrid, but we can assume that CL solutions are properly disinfecting the CL since only 1.1% of the positive PCR samples correspond to viable amoebae and, after four years, only one participant reported stronger ocular problems. Nevertheless, more studies are necessary to corroborate this hypothesis.  相似文献   

6.
Abstract

A study was conducted in the greenhouse to examine the resistance of three soybean cultivars against root-infecting fungi, and to determine the role of five strains of Pseudomonas aeruginosa in protecting the roots from these fungal pathogens. In this study soybean cv RAWAL was found to be less susceptible against charcoal rot fungus Macrophomina phaseolina than cvs PARC and BRAGG. Most of the strains of P. aeruginosa used as seed dressing significantly reduced M. phaseolina and Rhizoctonia solani infection on all three cvs PARC, BRAGG and RAWAL (p < 0.05). Most of the strains of P. aeruginosa were effective on cv PARC against Fusarium solani infection, while on cv BRAGG P. aeruginosa strain Pa3, and on cv RAWAL strain Pa5 were effective. Both strains Pa3 and Pa22 gave maximum plant height and fresh weight of shoots, respectively on cvs PARC and BRAGG than other strains. These characteristics make these P. aeruginosa strains good candidates for use as biocontrol agents against soil-borne plant pathogens.  相似文献   

7.
【背景】屎肠球菌为ESKAPE(由屎肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌属六大超级细菌的拉丁学名首字母组成)病原体之一,对多种抗菌药物具有耐药性,严重威胁全球人类健康,被世界卫生组织列入亟须研发新抗菌药的病原体名单。【目的】分离针对屎肠球菌的烈性噬菌体,测定其基本生物学特性并进行基因组测序分析,为屎肠球菌噬菌体疗法提供原料。【方法】从牧场污水中分离筛选出一株烈性屎肠球菌噬菌体,命名为Enterococcus phage 1A11,通过透射电镜观察噬菌体的形态,测定其最佳感染复数、一步生长曲线和裂解谱,并进行全基因组的测序和分析,以阐释该噬菌体的基本生物学特性。【结果】电镜下可观察到屎肠球菌噬菌体1A11具有典型的正二十面体头部结构和较长的尾部结构,属于有尾病毒目长尾病毒科,而且测得其最佳感染复数为0.01,裂解周期为70 min,潜伏期为30 min,暴发期为40 min,并特异性地对部分屎肠球菌产生裂解作用。噬菌体1A11的基因组大小为42 750 bp,GC含量为34.71%,含有70个推定的开放阅读框(open reading frame, O...  相似文献   

8.
【背景】嗜麦芽窄食单胞菌是一种广泛存在于医院和自然环境中的条件致病菌,其分离率与耐药率逐年增加。噬菌体是一类能特异性感染并杀灭细菌的病毒。【目的】分离一株新型嗜麦芽窄食单胞菌噬菌体,为临床嗜麦芽窄食单胞菌感染及防控提供补充手段。【方法】以临床分离的嗜麦芽窄食单胞菌为宿主菌,用点板法从医院污水中分离鉴定噬菌体;用双层平板法测定噬菌体效价及一步生长曲线等生物学特性;用透射电镜观察噬菌体形态;提取噬菌体基因组DNA进行全基因组测序,拼接噬菌体基因组并进行注释。【结果】分离到一株嗜麦芽窄食单胞菌裂解性噬菌体,命名为v B_Sma S_P11。该噬菌体感染宿主菌的潜伏期小于5 min,快速增殖60 min后达到平稳期,暴发量为100 PFU/cell。透射电镜观察该噬菌体为长尾噬菌体,具有典型的二十面体头和不可收缩的尾部。基因组测序结果表明,该噬菌体基因组全长44 600 bp,GC含量为63.7%,无抗生素耐受基因、毒力基因和t RNA,与NCBI数据库中所有已知嗜麦芽窄食单胞菌噬菌体相比同源性很低。基因组注释显示该噬菌体含有66个开放阅读框(open reading frame,ORF),其...  相似文献   

9.
Abstract

Microbial cells can rapidly form biofilm on endotracheal tubes (ETT) causing ventilator-associated pneumonia, a serious complication in patients receiving mechanical ventilation. A novel polyamide with a good balance of hydrophilic/hydrophobic moieties was used for the embedment of green-reduction silver nanoparticles (AgNPs) for the composite-coated ETT. The films were conformal with a thickness of ~ 17?±?3?µm accommodating high loading of 60?±?35?nm spherical-shaped AgNPs. The coated ETT resulted in a significant difference in reducing both planktonic growth and microbial adhesion of single and mixed-species cultures, compared with uncoated ETT (p?<?0.05). A time-kill assay demonstrated rapid bactericidal effects of the coating on bacterial growth and cell adhesion to ETT surface. Biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus, commonly encountered pathogens, was inhibited by > 96% after incubation for 72?h. Polyamide/AgNP composite-coated ETT provided a broad-spectrum activity against both Gram-positive and Gram-negative bacteria as well as Candida albicans and prolonged antimicrobial activity.  相似文献   

10.

This study proposed to investigate the effect of azurin on the major stages of pathogenesis (adhesion and invasion) of intestinal bacterial pathogens (Salmonella spp. and Escherichia coli) and epithelial pathogens (Staphylococcus aureus and Pseudomonas aeruginosa) on the human colorectal adenocarcinoma (Caco-2) cell line. Azurin protein was produced by cloning the azurin gene into pET21a and heterologous expression in E. coli BL21. The protein was purified using affinity chromatography and confirmed by Western blotting. The purified protein was evaluated by three experiments of adhesion and invasion assays, including exclusion, competition, and replacement. Azurin was observed to significantly inhibit the attachment and invasion of S. aureus, Salmonella spp., and E. coli, while no such inhibitory effects were observed on P. aeruginosa. In fact, the protein increased the adhesion of P. aeruginosa to the cell. In conclusion, our study proposes that azurin is a potential prophylactic or preventive helper candidate to inhibit the attachment and invasion of pathogenic bacteria to host cells and reduce the progression of the infection process. Our study also reveals the involvement of azurin in bacteria-host cell interactions, providing novel and important insights toward the elucidation of its biological function in this field. Thus, this study provides new opportunities to use azurin as an adjunct therapy against critical stages of infection by a wide range of pathogenic bacteria.

  相似文献   

11.
The flagellar protein (flagellin) was isolated and purified from strains of Pseudomonas aeruginosa, Burkholderia cepacia and Stenotrophomonas maltophilia. A significant difference was observed in the molecular weight of different flagellin preparations obtained from these bacterial isolates. Antiserum prepared against S. maltophilia flagellin did not react with flagellin of P. aeruginosa or/and B. cepacia on Immunoblot or in indirect ELISA. In addition the anti-flagellin did not agglutinate P. aeruginosa and B. cepacia. No inhibition of motility of P. aeruginosa and B. cepacia was observed in presence of antiserum; though the latter inhibited the motility of S. maltophilia. The results of the present study prove that no specific relationship existed among all the studied flagellar proteins obtained from closely related bacteria.  相似文献   

12.
Microbial substitution occasionally occurs following the administration of antimicrobials to horses that have pneumonia or pleuropneumonia. Four specific loop‐mediated isothermal amplification (LAMP) assays were developed to detect some equine respiratory pathogens, namely strains of the BacteroidesPrevotella group, Klebsiella pneumoniae, Stenotrophomonas maltophilia, and Staphylococcus aureus. These four LAMP assays and two previously published LAMP assays targeting Escherichia coli or Pseudomonas aeruginosa were used on clinical respiratory specimens and a high accordance found between the results of the LAMP assays and bacterial culture. Use of these LAMP assays could enable rapid detection of pathogenic bacteria and swift administration of the appropriate antimicrobials.  相似文献   

13.
【目的】查明引起湖北仙桃某中华鳖养殖场中华鳖发病死亡的病原及其特征。【方法】本研究分离患病中华鳖的病原,并结合形态特征、生理生化试验、16SrRNA基因鉴定,鉴定分离菌株;通过人工回归感染试验、药敏试验、全基因组测序与分析对分离菌株的致病性和耐药性进行研究;通过多位点序列分型(multilocus sequence type,MLST)分析,对分离菌株的流行情况进行探究。【结果】从患病中华鳖肝脏等部分分离到3株优势菌株HX8、FG10和GC20,16SrRNA基因同源性和生化特征分析鉴定为铜绿假单胞菌(Pseudomonas aeruginosa)。回归感染试验证实该菌株是引起本次中华鳖患病的病原菌。3株分离株的药敏实验结果基本一致,均对诺氟沙星、恩诺沙星等8种抗生素敏感,对氟苯尼考、多西环素、磺胺异恶唑等6种抗生素耐药。MLST鉴定表明,3株分离株均属于序列型(sequencetype,ST)252型,eBURST分析进一步显示ST252型与一些ST共同构成了克隆复合体(clonal complexes,CC) CC252,且ST252是CC252的原始序列型(founder ST)...  相似文献   

14.
Stenotrophomonas maltophilia is an emerging drug-resistant pathogen and an important opportunistic pathogen. S. maltophilia flagellin was purified using serial ultracentrifugation. The purity of flagellin was checked by SDS-PAGE. The antibodies were raised in rabbits. The presence of anti-flagellin and the titer of flagellin were detected by immunoblotting and bacterial agglutination techniques. Two methods (viable bacterial count and spectrophotometric methods) were applied to evaluate bacterial adhesion and biofilm formation. Pretreatment of S. maltophilia with dilutions of anti-flagellin (from 1/40 to 1/640) reduced the ability of S. maltophilia to adhere and form biofilms on polystyrene (P < 0.05). In the present study, the inhibition of bacterial adhesion to polystyrene was dose-dependent. The positive correlation was observed between the antibody dilutions and bacterial adhesion (CFU/mL) (r > +0.5, P < 0.05), while, the negative correlation (r < ?0.5, P < 0.05) was observed between the percentage of adhesion inhibition and anti-flagellin dilutions. The current study proved the direct role of S. maltophilia flagellin in bacterial adhesion to and biofilm formation on polystyrene.  相似文献   

15.
A rapid identification method of glucose nonfermentative gram-negative rods was established and 320 strains isolated were divided into five groups according to their characteristics in pigmentation, acid from glucose, cytochrome oxidase activity and motility. Further characterization of the strains in each group resulted in the identification that the strains in group I were Pseudomonas aeruginosa, strains in group II, P. aeruginosa and Pseudomonas putida. Achromogenic strains of P. aeruginosa were classified into group III, Pseudomonas maltophilia, Pseudomonas alcaligenes and Alcaligenes faecalis into group IV and Acinetobacter calcoaceticus (Acinetobacter anitratus and Achromobacter lwoffii) in group V. When fluorescent pigment production was taken as a standard, 259 out of 263 chromogenic strains were identified as P. aeruginosa and the remaining four were P. putida. Whereas forty-five achromogenic strains included twenty-four A. calcoaceticus, eight P. aeruginosa, six A. faecalis, five P. maltophilia and two P. alcaligenes. From May 1970 to June 1971, 368 strains of glucose nonfermentative rods were isolated from clinical specimens sent to the Central Laboratories of Tohoku University Hospital and three fourth (286/368) of the isolates were P. aeruginosa  相似文献   

16.
In order to investigate the effect of moxifloxacin on survival, lipid peroxidation and inflammation in immunosuppressed rats with soft tissue infection caused by Stenotrophomonas maltophilia, 144 white male Wistar rats were randomized into six groups: Groups A and B received saline or moxifloxacin once per day, respectively; Groups C and D received saline or moxifloxacin twice per day, respectively, and Groups E and F received saline or moxifloxacin three times per day, respectively. Blood samples were taken at 6 and 30 hr after administration of S. maltophilia. Malonodialdehyde (MDA), WBC counts, bacterial tissue overgrowth, serum concentrations of moxifloxacin and survival were assessed. Survival analysis proved that treatment with moxifloxacin every 8 hr was accompanied by longer survival than occurred in any other group. Tissue cultures 30 hr after bacterial challenge showed considerably less bacterial overgrowth in the spleens and lungs of moxifloxacin‐treated than in saline‐treated animals, but not in their livers. At 6 hr there were no statistically significant differences between groups. However, at 30 hr, MDA concentrations were significantly greater (P = 0.044) and WBC counts significantly lower (P = 0.026) in group D than in group C. No statistically significant variations were observed between the other groups. Moxifloxacin possibly stimulates lipid peroxidation and enhances phagocytosis, as indicated by MDA production and survival prolongation, without being toxic, as indicated by WBC count. Therefore, under the appropriate conditions, moxifloxacin has a place in treatment of infections in immunosuppressed patients and of infections caused by S. maltophilia.  相似文献   

17.
The growth conditions are known to influence the bacterial adhesion to different kinds of surfaces. In the present study the adhering ability of S. maltophilia, on growth in nutrient rich media (Tryptic Soy Broth (TSB)) and minimal media (Luria Bertani (LB)) was checked by viable cell count and spectrophotometric method. TSB grown S. maltophilia showed higher adhesion compared to bacteria grown in LB broth, to both biotic and abiotic surfaces. However, when bacteria were grown in LB broth supplemented with different concentrations of glucose, under aerobic conditions, the bacteria grown at lower glucose concentration (2 gm/l) showed maximumadhesion to abiotic surfaces (polystyrene microliter plate) compared to biotic surfaces (mouse trachea, mouse tracheal mucus and HEp-2 cells line). Maximum adhesion to biotic surfaces was seen with cells grown at 4 gm/l of glucose concentration. On the contrary if the cell was grown under microaerophilic conditions maximum adhesion to abiotic and biotic surfaces was achieved with bacteria grown at 1 gm/l and 2 gm/l of glucose concentration respectively. A negative correlation was observed between glucose concentrations and pH of media, the latter declined faster under microaerophilic conditions as compared to aerobic condition.  相似文献   

18.

The mutual influences of Pseudomonas aeruginosa PAO1 and Desulfovibrio desulfuricans subsp. desulfuricans (ATCC 29577) on their adhesion to stainless steel were investigated in batch and column experiments. It was found that P. aeruginosa promoted the adhesion of D. desulfuricans under conditions of turbulence, but not under quiescent conditions. The enhancement involved the alignment of most D. desulfuricans along P. aeruginosa cells and was attributed to the additional interaction surface area provided by adhered P. aeruginosa to aligning D. desulfuricans cells. A slightly positive effect of pre-adhered D. desulfuricans on the adhesion of P. aeruginosa was found. Under condition of laminar flow, substantially better adhesion of D. desulfuricans to confluent P. aeruginosa biofilms than to steel was observed. The mutual influences are discussed in terms of more favorable adhesion energies and the influence of changed hydraulic conditions due to the roughness of P. aeruginosa biofilms.  相似文献   

19.

Introduction

Cystic fibrosis (CF) airways are colonized by a polymicrobial community of organisms, termed the CF microbiota. We sought to define the microbial constituents of the home environment of individuals with CF and determine if it may serve as a latent reservoir for infection.

Methods

Six patients with newly identified CF pathogens were included. An investigator collected repeat sputum and multiple environmental samples from their homes. Bacteria were cultured under both aerobic and anaerobic conditions. Morphologically distinct colonies were selected, purified and identified to the genus and species level through 16S rRNA gene sequencing. When concordant organisms were identified in sputum and environment, pulsed-field gel electrophoresis (PFGE) was performed to determine relatedness. Culture-independent bacterial profiling of each sample was carried out by Illumina sequencing of the V3 region of the 16s RNA gene.

Results

New respiratory pathogens prompting investigation included: Mycobacterium abscessus(2), Stenotrophomonas maltophilia(3), Pseudomonas aeruginosa(3), Pseudomonas fluorescens(1), Nocardia spp.(1), and Achromobacter xylosoxidans(1). A median 25 organisms/patient were cultured from sputum. A median 125 organisms/home were cultured from environmental sites. Several organisms commonly found in the CF lung microbiome were identified within the home environments of these patients. Concordant species included members of the following genera: Brevibacterium(1), Microbacterium(1), Staphylococcus(3), Stenotrophomonas(2), Streptococcus(2), Sphingomonas(1), and Pseudomonas(4). PFGE confirmed related strains (one episode each of Sphinogomonas and P. aeruginosa) from the environment and airways were identified in two patients. Culture-independent assessment confirmed that many organisms were not identified using culture-dependent techniques.

Conclusions

Members of the CF microbiota can be found as constituents of the home environment in individuals with CF. While the majority of isolates from the home environment were not genetically related to those isolated from the lower airways of individuals with CF suggesting alternate sources of infection were more common, a few genetically related isolates were indeed identified. As such, the home environment may rarely serve as either the source of infection or a persistent reservoir for re-infection after clearance.  相似文献   

20.
作为人类条件性感染的前三大病原菌之一的铜绿假单胞菌,是一种革兰氏阴性细菌,对免疫功能低下和囊性纤维化患者可以造成严重和持续性感染。造成这种持续感染的原因主要是由于细菌接收外界信号后,在自身调控网络的协同作用下,会依附于固体表面,并产生胞外多糖、基质蛋白和胞外DNA等大分子物质形成高度结构化的膜状复合物将自身包裹形成生物被膜群体结构。生物被膜可以有效帮助细菌定殖、提高细菌对抗菌物质和宿主免疫反应的抵抗能力、促进群落细菌的细胞-细胞之间的信号交流等,是临床治疗中病原菌慢性感染和反复感染最重要的原因之一。本篇综述重点介绍了铜绿假单胞菌生物被膜的各组成成分及其在生物被膜形成中的重要功能,并进一步阐述了群体感应系统(las、rhl、pqs与iqs)和c-di-GMP对铜绿假单胞菌生物被膜形成的调控作用。通过本篇综述可以更清晰地了解细菌生物被膜形成和调控的过程,为开发新的治疗生物被膜感染策略提供帮助。  相似文献   

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