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At least 739 records · Page 41Linked to original sources
PubMed2004

Faucets as a reservoir of endemic Pseudomonas aeruginosa colonization/infections in intensive care units.

OBJECTIVE: To evaluate the role of faucets as a reservoir for Pseudomonas aeruginosa colonization/infection of patients hospitalized in intensive care units (ICUs). DESIGN: Prospective epidemiological investigation performed during a nonepidemic period of 1 year. The inner part of the ICU faucets were swabbed for P. aeruginosa. Data were recorded on all patients with at least one culture of a clinical specimens positive for P. aeruginosa. Pulsed-field gel electrophoresis was used to characterize the strains. SETTING: Five ICUs of a university hospital which are supplied by two separate water distribution networks. PATIENTS: During a 1-year period 132 cases were investigated. RESULTS: In 42% of cases (56/132) there were isolates identical to those found in the faucets, with a total of nine different genotypes. Among the nine genotypes isolated from both patients and faucets one of them, the most prevalent, was isolated in the two networks and in 30 cases. The other eight genotypes were recovered almost exclusively from either one (three genotypes in 12 cases) or the other (five genotypes in 12 cases) network and from the patients in the corresponding ICUs. CONCLUSIONS: These results suggest that the water system of the ICUs was the primary reservoir of patient's colonization/infection with P. aeruginosa in a substantial proportion of patients, although the exact mode of acquisition could not be determined.

Colony Count, Microbial↗
PubMed2024

Recent advances in environmental antibiotic resistance genes detection and research focus: From genes to ecosystems.

Antibiotic resistance genes (ARGs) persistence and potential harm have become more widely recognized in the environment due to its fast-paced research. However, the bibliometric review on the detection, research hotspot, and development trend of environmental ARGs has not been widely conducted. It is essential to provide a comprehensive overview of the last 30 years of research on environmental ARGs to clarify the changes in the research landscape and ascertain future prospects. This study presents a visualized analysis of data from the Web of Science to enhance our understanding of ARGs. The findings indicate that solid-phase extraction provides a reliable method for extracting ARG. Technological advancements in commercial kits and microfluidics have facilitated the efficacy of ARGs extraction with significantly reducing processing times. PCR and its derivatives, DNA sequencing, and multi-omics technology are the prevalent methodologies for ARGs detection, enabling the expansion of ARG research from individual strains to more intricate microbial communities in the environment. Furthermore, due to the development of combination, hybridization and mass spectrometer technologies, considerable advancements have been achieved in terms of sensitivity and accuracy as well as lowering the cost of ARGs detection. Currently, high-frequency terms such as "Antibiotic Resistance, Antibiotics, and Metagenomics" are the center of attention for study in this area. Prominent topics include the investigation of anthropogenic impacts on environmental resistance, as well as the dynamics of migration, dissemination, and adaptation of environmental ARGs, etc. The research on environmental ARGs has made significant advancements in the fields of "Microbiology" and "Biotechnology Applied Microbiology". Over the past decade, there has been a notable increase in the fields of "Environmental Sciences Ecology" and "Engineering" with a similar growth trend observed in "Water Resources". These three domains are expected to continue driving extensive study within the realm of environmental ARGs.

Drug Resistance, Microbial↗
PubMed2006

Biofilms: implications in bioremediation.

Biofilms are assemblages of single or multiple populations that are attached to abiotic or biotic surfaces through extracellular polymeric substances. Gene expression in biofilm cells differs from planktonic stage expression and these differentially expressed genes regulate biofilm formation and development. Biofilm systems are especially suitable for the treatment of recalcitrant compounds because of their high microbial biomass and ability to immobilize compounds. Bioremediation is also facilitated by enhanced gene transfer among biofilm organisms and by the increased bioavailability of pollutants for degradation as a result of bacterial chemotaxis. Strategies for improving bioremediation efficiency include genetic engineering to improve strains and chemotactic ability, the use of mixed population biofilms and optimization of physico-chemical conditions. Here, we review the formation and regulation of biofilms, the importance of gene transfer and discuss applications of biofilm-mediated bioremediation processes.

Bacteria↗
PubMed2003

Modulation of Rab5 and Rab7 recruitment to phagosomes by phosphatidylinositol 3-kinase.

Phagosomal biogenesis is central for microbial killing and antigen presentation by leukocytes. However, the molecular mechanisms governing phagosome maturation are poorly understood. We analyzed the role and site of action of phosphatidylinositol 3-kinases (PI3K) and of Rab GTPases in maturation using both professional and engineered phagocytes. Rab5, which is recruited rapidly and transiently to the phagosome, was found to be essential for the recruitment of Rab7 and for progression to phagolysosomes. Similarly, functional PI3K is required for successful maturation. Remarkably, inhibition of PI3K did not preclude Rab5 recruitment to phagosomes but instead enhanced and prolonged it. Moreover, in the presence of PI3K inhibitors Rab5 was found to be active, as deduced from measurements of early endosome antigen 1 binding and by photobleaching recovery determinations. Though their ability to fuse with late endosomes and lysosomes was virtually eliminated by wortmannin, phagosomes nevertheless recruited a sizable amount of Rab7. Moreover, Rab7 recruited to phagosomes in the presence of PI3K antagonists retained the ability to bind its effector, Rab7-interacting lysosomal protein, suggesting that it is functionally active. These findings imply that (i) dissociation of Rab5 from phagosomes requires products of PI3K, (ii) PI3K-dependent effectors of Rab5 are not essential for the recruitment of Rab7 by phagosomes, and (iii) recruitment and activation of Rab7 are insufficient to induce fusion of phagosomes with late endosomes and lysosomes. Accordingly, transfection of constitutively active Rab7 did not bypass the block of phagolysosome formation exerted by wortmannin. We propose that Rab5 activates both PI3K-dependent and PI3K-independent effectors that act in parallel to promote phagosome maturation.

Adaptor Proteins, Signal Transducing↗
PubMed2002

Bacteriological qualities of indoor and out-door drinking water in Kibera sub-location of Nairobi, Kenya.

OBJECTIVES: To compare the bacteriological quality of out-house (tank or standpipe) water and in-house drinking water (storage containers) and determine the risk factors influencing it. DESIGN: A cross-sectional study. SETTING: The study was carried out in Kibera slums located 7 km southwest from the Nairobi City centre. SUBJECTS: Water samples from twenty outside tanks/standpipes and sixty from in-house water storage containers. MAIN OUTCOME MEASURES: Pour plate method was used to enumerate total bacterial counts in water, while the multiple tube technique was used to determine faecal coliform (FC) and faecal streptococci (FS) numbers. A questionnaire and environmental observation were used to determine the risk factors influencing bacteriological quality of water. RESULTS: The mean total bacterial counts (TBC) for out-house water was 46.6 per 100 ml while that for in-house water was 818.2 per 100 ml. Faecal coliforms were isolated from 7 (35%) standpipes and 57 (95%) in-house storage containers. The mean faecal coliform count was 93 and 103.4 per 100 ml for out-house and in-house water, respectively. The counts were significantly higher in the latter. Faecal streptococci were isolated from 2 (10%) standpipes and 37 (61.7%) in-house storage containers. The mean faecal streptococci counts were 35 and 65 per 100 ml for out-house and in-house water sources, respectively. Escherichia coli was isolated in 2 (10%) of out-house water and 30 (50%) of in-house. Of these, four were enteropathogenic, serotype 011 from one out-house water source and serotypes 011, 011, 0112ac from in-house water sources. CONCLUSIONS: Bacteriological contamination of water at the source with a further deterioration between the collection points and homes was observed. A defective water delivery system and inadequate environmental sanitation were a potential source of contamination for out-house water. Scoops were a major source of contamination for stored water.

Colony Count, Microbial↗
PubMed2002

Therapeutic challenges posed by bacterial bioterrorism threats.

The events of the autumn of 2001 in the United States made it clear that the spectre of the use of microorganisms to intentionally harm humans is a reality. The current strategy to control disease outbreaks caused by the intentional release of bacteria is to use antimicrobial agents, both therapeutically and prophylactically. However, multidrug-resistant strains of bacterial bioterrorism agents occur naturally or have been bio-engineered, indicating how vulnerable this strategy is.

Anthrax↗
PubMed2003

Effect of trehalose on survival of Bradyrhizobium japonicum during desiccation.

AIMS: A major reason for the ineffectiveness of legume inoculants in the field is the rapid death of rhizobia because of desiccation. The major purpose of this study was to identify conditions under which alpha,alpha-trehalose would improve survival of Bradyrhizobium japonicum during desiccation. METHODS AND RESULTS: Trehalose was added to cultures just prior to desiccation or was supplied to bacteria during the 6-day growth period. A wide variety of trehalose concentrations was tested. Trehalose added to cultures at the time of desiccation improved survival slightly, but trehalose loading during growth was much more effective in protection against desiccation. Growth of bacteria with 3 mmol l-1 trehalose increased trehalose concentration in cells by about threefold and increased survival of cells placed on soya bean [Glycine max (L.) Merr.] seeds by two- to four-fold after 2 or 24 h. Average of overall results indicate that growth of bacteria with trehalose in the medium resulted in a 294% increase in survival after 24 h of desiccation. The concentration of trehalose in cells was very highly correlated with survival of bacteria. When trehalose-loaded cells were suspended in buffer or water, 60-85% of cellular trehalose was lost in about 1 h and, in spite of these losses, survival during desiccation was not reduced. CONCLUSIONS: Accumulation of trehalose in the cytoplasm is critical to the survival of B. japonicum during desiccation. Increasing the periplasmic concentration of trehalose is also beneficial but is not so critical as the concentration of trehalose in the cytoplasm. Because B. japonicum cannot utilize trehalose as a carbon source, cells can be loaded with trehalose by providing the disaccharide during the growth period. SIGNIFICANCE AND IMPACT OF THE STUDY: Although it may not be practical to use trehalose as a carbon source in inoculant production, it may be possible to engineer greater trehalose accumulation in rhizobia. Trehalose concentration in cells should be a useful predictor of survival during desiccation.

Bacteriological Techniques↗
PubMed1992

Introducing mutations into the single-copy chromosomal 23S rRNA gene of the archaeon Halobacterium halobium by using an rRNA operon-based transformation system.

A vector-transformation system is described that permits replacement of a portion of the single rRNA operon of the archaeon Halobacterium halobium with a homologous fragment from a vector-borne gene. The vector construct contains three functional sections: (i) an entire H. halobium rRNA operon with two selective mutations in the 23S rRNA gene, the substitutions of A----G at position 1159 conferring resistance to thiostrepton and C----U at position 2471 conferring resistance to anisomycin; (ii) the complete pHSB1 plasmid from Halobacterium sp. SB3, which interferes with vector maintenance in the transformed halobacterial cells; and (iii) a segment of the pBR322 plasmid that permits vector replication in Escherichia coli. Transformation of H. halobium with the vector plasmid generates cells resistant to both anisomycin and thiostrepton that can be selected for, and discriminated from spontaneous mutants, by a two-step selection procedure. After transformation, the plasmid recombines homologously with the chromosome so that the plasmid-borne rDNA segment with resistance markers substitutes for the corresponding region of the chromosomal rRNA operon, and the transforming plasmid is lost. Eventually, this leads to a homogeneous population of the mutant ribosomes in the cell. Other mutations that are engineered in the vector-borne rRNA sequences can be transferred to the chromosomal rRNA operon concomitantly with the selective markers. The system has considerable potential for ribosomal engineering.

Anisomycin↗
PubMed1988

In situ survival of plasmid-bearing and plasmidless Pseudomonas aeruginosa in pristine tropical waters.

Two rare wild-type strains of Pseudomonas aeruginosa were mixed in membrane diffusion chambers and then introduced into a natural freshwater environment for 72 h. The plasmid-containing strain (R serotype 15) and the plasmidless strain (H serotype 5) had initial bacterial densities of 2 x 10(5) cells per ml. Samples collected from the chambers were analyzed for viable and direct counts and for acquired-resistance frequencies. Suspected transconjugant-to-donor ratios ranged from 0.5 to 1.3; transfer percentages ranged from 13 to 70%. [3H]thymidine uptake indicated DNA synthesis in both strains as well as in transconjugants. These studies indicate that rare wild-type bacterial strains with large plasmid loads can survive as well as can bacteria with low plasmid loads when exposed to the in situ conditions of a tropical freshwater habitat. These results also suggest that genetic modification of indigenous microbiota through conjugation or transformation is feasible when rare wild-type strains or genetically engineered microorganisms are released in large numbers in tropical aquatic ecosystems.

Colony Count, Microbial↗
PubMedDate not supplied

Genetic engineering of structural protein polymers.

Genetic and protein engineering are components of a new polymer chemistry that provide the tools for producing macromolecular polyamide copolymers of diversity and precision far beyond the current capabilities of synthetic polymer chemistry. The genetic machinery allows molecular control of chemical and physical chain properties. Nature utilizes this control to formulate protein polymers into materials with extraordinary mechanical properties, such as the strength and toughness of silk and the elasticity and resilience of mammalian elastin. The properties of these materials have been attributed to the presence of short repeating oligopeptide sequences contained in the proteins, fibroin, and elastin. We have produced homoblock protein polymers consisting exclusively of silk-like crystalline blocks and elastin-like flexible blocks. We have demonstrated that each homoblock polymer as produced by microbial fermentation exhibits measurable properties of crystallinity and elasticity. Additionally, we have produced alternating block copolymers of various amounts of silk-like and elastin-like blocks, ranging from a ratio of 1:4 to 2:1, respectively. The crystallinity of each copolymer varies with the amount of crystalline block interruptions. The production of fiber materials with custom-engineered mechanical properties is a potential outcome of this technology.

Amino Acid Sequence↗
PubMed2002

Enhancement of a two-phase partitioning bioreactor system by modification of the microbial catalyst: demonstration of concept.

Application of two-phase partitioning bioreactors (TPPB) to the degradation of phenol and xenobiotics has been limited by the fact that many organic compounds that would otherwise be desirable delivery solvents can be utilized by the microorganisms employed. The ability to metabolize the solvent itself could interfere with xenobiotic degradation, limiting remediation efficiency, and hence represents a microbial characteristic incompatible with process goals. To avoid the issue of bioavailability, previous TPPB applications have relied on complex and often expensive delivery solvents or suboptimal catalyst-solvent pairings. In an effort to enhance TPPB activity and applicability, a genetically engineered derivative of Pseudomonas putida ATCC 11172 mutated in its ability to utilize medium-chain-length alcohols was generated (AVP2) and applied as the catalyst within a TPPB system with decanol as the delivery solvent. Kinetic analysis verified that the genetic alteration had not negatively affected phenol degradation. The volumetric productivity of AVP2 (0.48 g/L x h(-1)) was equivalent to that seen for wild-type ATCC 11172 (0.51 g/L x h(-1)), but a comparison of initial cell concentrations and yields revealed an improved phenol-degrading efficiency for the mutant under process conditions. Yield coefficients, cell dry weight, and viable count determinations all confirmed the stability of the modified phenotype. This work illustrates the possibilities for TPPB process enhancement through a careful combination of genetic modification and solvent selection.

Alcohols↗
PubMed2002

Biosynthesis of novel thermoplastic polythioesters by engineered Escherichia coli.

The development of non-petrochemical sources for the plastics industry continues to progress as large multinationals focus on renewable resources to replace fossil carbon. Many bacteria are known to accumulate polyoxoesters as water-insoluble granules in the cytoplasm. The thermoplastic and/or elastomeric behaviour of these biodegradable polymers holds promise for the development of various technological applications. Here, we report the synthesis and characterization of microbial polythioesters (PTEs), a novel class of biopolymers of general technological relevance. Biosynthesis of PTE homopolymers was achieved using a recombinant strain of Escherichia coli that expressed a non-natural pathway consisting of a butyrate kinase, a phosphotransbutyrylase, and a PHA synthase. Different homopolymers were produced, consisting of either 3-mercaptopropionate, 3-mercaptobutyrate, or 3-mercaptovalerate repeating units, if the respective mercaptoalkanoic acids were provided as precursor substrates to the fermentative process. The PTEs contributed up to 30% (w/w) of the cellular dry weight and were identified as hydrophobic inclusions in the cytoplasm. The chemical and stereochemical homogeneity of the purified PTEs were identified by different methods, and the estimated physical properties were compared to the oxypolyester equivalents, revealing low crystalline order and, for the poly(3-mercaptopropionate) improved thermal stability. The ability to produce PTEs through a biosynthetic route opens up new avenues in the field of biomaterials.

Biopolymers↗
PubMed2004

Microbial hyaluronic acid production.

Hyaluronic acid (HA) is a commercially valuable medical biopolymer increasingly produced through microbial fermentation. Viscosity limits product yield and the focus of research and development has been on improving the key quality parameters, purity and molecular weight. Traditional strain and process optimisation has yielded significant improvements, but appears to have reached a limit. Metabolic engineering is providing new opportunities and HA produced in a heterologous host is about to enter the market. In order to realise the full potential of metabolic engineering, however, greater understanding of the mechanisms underlying chain termination is required.

Animals↗
PubMed1996

Biological control of plant root pathogens.

Rhizobacteria introduced to control soil-borne root diseases must establish metabolically active populations that mediate protection either by direct antagonism of pathogens or by stimulation of host plant defenses. Recent interest has focused on the genetic and biochemical basis of disease control and the influence of environmental factors on the expression and activity of biocontrol mechanisms. The cloning and sequencing of genes involved in the production of microbial metabolites playing key roles in plant defense opens new possibilities for improving the performance of biocontrol agents.

Anti-Bacterial Agents↗
PubMed2004

[Study on the reconstitution in vitro and photochemical activities of phytochrome from the Synechocystis sp. PCC6803].

Genomic DNA sequence analysis of phytochrome like photoreceptors in a number of bacteria revealed several open reading frames (ORFs) encoding proteins with amino acid sequences homologous to plant phytochromes. The phytochrome like photoreceptors, collectively called bacteriophytochromes, contain an N-terminal domain homologous to the chromophore-binding domain (CBD) of higher plants and a C-terminal domain of histidine kinase domain( HKD). Due to their simple structure, bacteriophytochromes broaden the view of phytochrome evolution and provide us with a simple model to investigate phytochrome-mediated light signal in higher plants. In this report, the bacteriophytochromes from Synechocystis sp. PCC6803 were investigated. The gene cph1 and its fragment cph1 (C-435) were isolated from the Synechocystis sp. PCC6803 genomic DNA by polymerase chain reaction(PCR) using specific primers. Then, the genes were cloned with the vector pBluescript, yielding plasmids pBlu-cphl and pBlu-cph1 ( C-435), before they are subcloned with the vector pET30, using the EcoRV and Xho I restriction sites. pBlu-cph1, pBlu-cph1 (N-435) were cleaved with Sma I and Xho I, and the released genes were ligated to the pET30a fragment. The E. coli [strain BL21 (DE3)] cells containing recombinant pET30a were grown in medium RB at 20 degrees C, and harvested 6 h later after induction with isopropyl thio-beta-D-galactoside (IPTG). Then, reconstitution systems were employed to study the characteristics of the genes. In the reconstitution system, autoassembly of aprotein of phytochrome with PCB was investigated. The chromophore addition was an autocatalytic process. Reconstitution products were red/infrared (R/FR) photochromic, which was similar to that of the phytoehrome in higher plants. How ever, the spectral change ratios (deltaAmax/deltaAmin) of the two fragments differed from each other. It was also shown that PCB was covalently bound to apo-protein via Zn2+ fluoresc ence SDS-PAGE. After irradiation by light of 700 nm, the maximum absorption spectrum o f holo-Cphl was 650nm. The absorption of it after denaturatior in the dark with ur ea in the presence of hydrochloric acid (pH = 2) was 660nm, which was similar with th at of cis-PCB. In addition, after irradiation by light of 650nm, the maximum absorption spectrum of holo-Cph1 was 700nm. The absorption of it after denaturation in the dark with urea in the presence of hydrochloric acid (pH = 2) was 600nm, which was similar with that of trans-PCB. The result showed that the photochromism of phytochrome resulted from the isomerizaation of chromophore (PCB in this report). The reconstitution of Cph1 (C-435) under the same condition supported the conclusion. Fluorescence emission spectrum of the products suggested that bacteriophytochrom e structure with cis-PCB was more stable than that with trans-PCB. The new reconstitution system in this report sets a base for the application of phytochrome as photochromic biomaterials in biosensors. In addition, phytochrome shows great potential in food, cosmetic and biological engineering, etc.

Bacterial Proteins↗
PubMed2006

The chemistry of conventional and alternative treatment systems for the neutralization of acid mine drainage.

The oxidation of pyritic mining waste is a self-perpetuating corrosive process which generates acid mine drainage (AMD) effluent for centuries or longer. The chemical neutralization of these complex, buffered effluents result in unstable, metal-laden sludges, which require disposal to minimize long-term environmental consequences. A variety of passive treatment systems for AMD, developed in the past two decades, combine limestone and organic substrates in constructed wetlands. These systems work well initially but over the longer term fail due to clogging with and the depletion of available organic carbon. However, some ecologically engineered systems, which exploit the activities of acid reducing microbes in the sediment, rely on photosynthesis in the water column as a source of organic matter. The primary productivity in the water column, which also generates some alkalinity, provides electron donors for the microbial reduction processes in the sediment. In its consideration of 'passive' systems, the literature has placed undue emphasis on sulphate reduction; thermodynamical iron reduction is equally important as is the need to prevent iron oxidation. Secondary precipitates of iron play a significant role in sediment-driven biomineralization processes, which affect the anaerobic degradation of organic matter and the stability of the resulting metal sulfides. One such passive system, which utilized a floating root mass as a source of organic carbon, is described. An extensive review of the literature and the chemical and biogeochemical reactions of AMD treatment systems, lead to the conclusion, that sediment based ecological systems offer the greatest potential for the sustainable treatment of AMD.

Biodegradation, Environmental↗
PubMed2002

Identification and analysis of a bottleneck in PCB biodegradation.

The microbial degradation of polychlorinated biphenyls (PCBs) provides the potential to destroy these widespread, toxic and persistent environmental pollutants. For example, the four-step upper bph pathway transforms some of the more than 100 different PCBs found in commercial mixtures and is being engineered for more effective PCB degradation. In the critical third step of this pathway, 2,3-dihydroxybiphenyl (DHB) 1,2-dioxygenase (DHBD; EC 1.13.11.39) catalyzes aromatic ring cleavage. Here we demonstrate that ortho-chlorinated PCB metabolites strongly inhibit DHBD, promote its suicide inactivation and interfere with the degradation of other compounds. For example, k(cat)(app) for 2',6'-diCl DHB was reduced by a factor of approximately 7,000 relative to DHB, and it bound with sufficient affinity to competitively inhibit DHB cleavage at nanomolar concentrations. Crystal structures of two complexes of DHBD with ortho-chlorinated metabolites at 1.7 A resolution reveal an explanation for these phenomena, which have important implications for bioremediation strategies.

Binding Sites↗
PubMed2000

Strategy for systematic assembly of large RNA and DNA genomes: transmissible gastroenteritis virus model.

A systematic method was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in nature. The approximately 30-kb genome, coupled with regions of genomic instability, has hindered the development of a full-length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine. Using a novel approach, six adjoining cDNA subclones that span the entire TGEV genome were isolated. Each clone was engineered with unique flanking interconnecting junctions which determine a precise systematic assembly with only the adjacent cDNA subclones, resulting in an intact TGEV cDNA construct of approximately 28.5 kb in length. Transcripts derived from the full-length TGEV construct were infectious, and progeny virions were serially passaged in permissive host cells. Viral antigen production and subgenomic mRNA synthesis were evident during infection and throughout passage. Plaque-purified virus derived from the infectious construct replicated efficiently and displayed similar plaque morphology in permissive host cells. Host range phenotypes of the molecularly cloned and wild-type viruses were similar in cells of swine and feline origin. The recombinant viruses were sequenced across the unique interconnecting junctions, conclusively demonstrating the marker mutations and restriction sites that were engineered into the component clones. Full-length infectious constructs of TGEV will permit the precise genetic modification of the coronavirus genome. The method that we have designed to generate an infectious cDNA construct of TGEV could theoretically be used to precisely reconstruct microbial or eukaryotic genomes approaching several million base pairs in length.

Animals↗