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At least 991 records · Page 55Linked to original sources
PubMed1983

Radioassay for hydrogenase activity in viable cells and documentation of aerobic hydrogen-consuming bacteria living in extreme environments.

An isotopic tracer assay based on the hydrogenase-dependent formation of tritiated water from tritium gas was developed for in life analysis of microbial hydrogen transformation. This method allowed detection of bacterial hydrogen metabolism in pure cultures or in natural samples obtained from aquatic ecosystems. A differentiation between chemical-biological and aerobic-anaerobic hydrogen metabolism was established by variation of the experimental incubation temperature or by addition of selective inhibitors. Hydrogenase activity was shown to be proportional to the consumption or production of hydrogen by cultures of Desulfovibrio vulgaris, Clostridium pasteurianum, and Methanosarcina barkeri. This method was applied, in connection with measurements of free hydrogen and most-probable-number enumerations, in aerobic natural source waters to establish the activity and document the ecology of hydrogen-consuming bacteria in extreme acid, thermal, or saline environments. The utility of the assay is based in part on the ability to quantify bacterial hydrogen transformation at natural hydrogen partial pressures, without the use of artificial electron acceptors.

Journal Article↗
PubMed1994

Improved Method for Recovery of mRNA from Aquatic Samples and Its Application to Detection of mer Expression.

Previously described methods for extraction of mRNA from environmental samples may preclude detecting transcripts from genes that were present in low abundance in aquatic bacterial communities. By combining a boiling sodium dodecyl sulfate-diethylpyrocarbonate lysis step with acid-guanidinium extraction, we improved recovery of target mRNA from both pure cultures and environmental samples. The most significant advantage of the new protocol is that it is easily adapted to yield high recovery of mRNA from 142-mm-diameter flat filters and high-capacity cartridge filters. The lysis and extraction procedures are more rapid than previously described methods, and many samples can be handled at once. RNA extracts have been shown to be free of contaminating DNA. The lysis procedure does not damage target mRNA sequences, and mRNA can be detected from fewer than 10 bacterial cells. We used the new method to examine transcripts of genes responsible for detoxification of mercurial compounds. Induction of merA (specifying mercuric reductase) transcripts in stationary-phase Pseudomonas aeruginosa containing Tn501 occurred within 60 s of HgCl(2) addition and was proportional to the amount of Hg(II) added. The new technique also allowed the detection of merA transcripts from the microbial community of a mercury-contaminated pond (Reality Lake, Oak Ridge, Tenn.). Significant differences in merA transcript abundance were observed between different locations associated with the lake. The results indicate that the new method is simple and rapid and can be applied to the study of mer gene expression of aquatic communities in their natural habitats.

Journal Article↗
PubMed2004

Novel sulfonolipid in the extremely halophilic bacterium Salinibacter ruber.

Salinibacter ruber is an extremely halophilic bacterium, phylogenetically affiliated with the Flavobacterium/Cytophaga branch of the domain Bacteria. Electrospray mass analyses (negative ion) of the total lipid extract of a pure culture of S. ruber shows a characteristic peak at m/z 660 as the most prominent peak in the high-mass range of the spectrum. A novel sulfonolipid, giving rise to the molecular ion [M-H]- of m/z 660, has been identified. The sulfonolipid isolated and purified by thin-layer chromatography was shown by chemical degradation, mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance analysis to have the structure 2-carboxy-2-amino-3-O-(13'-methyltetradecanoyl)-4-hydroxy-18-methylnonadec-5-ene-1-sulfonic acid. This lipid represents about 10% of total cellular lipids, and it appears to be a structural variant of the sulfonolipids found as main components of the cell envelope of gliding bacteria of the genus Cytophaga and closely related genera (W. Godchaux and E. R. Leadbetter, J. Bacteriol. 153:1238-1246, 1983) and of diatoms (R. Anderson, M. Kates, and B. E. Volcani, Biochim. Biophys. Acta 528:89-106, 1978). Since this sulfonolipid has never been observed in any other extreme halophilic microorganism, we consider the peak at m/z 660 the lipid signature of Salinibacter. This study suggests that this novel sulfonolipid may be used as a chemotaxonomic marker for the detection of Salinibacter within the halophilic microbial community in saltern crystallizer ponds and other hypersaline environments.

Bacteroidetes↗
PubMed1974

Microbial metabolism and dynamic changes in the electrical conductivity of soil solutions: a method for detecting extraterrestrial life.

The addition of 0.5% glucose solutions to 12 different air-dried soils always resulted in increased electrical conductivity and water-soluble Ca and Mg in the soil solutions. The kinetics and magnitude of these changes for at least two and usually all three of these parameters over a 14-day period were clearly distinguishable from the changes in heat-sterilized controls or unsterilized controls without added glucose. In general, maximal values were achieved more rapidly under aerobic than anaerobic incubation. Some soils (less than half) also showed significant increases in water-soluble Na or K when compared with the controls. The 12 different soils studied represented four general soil groups: I, leached acid upland soils; II, saline alkaline soils; III, nonsaline neutral soils; and IV, high organic soils. Viable counts ranged from 10(4) to 10(7) per cm(3) of air-dried soil. Glucose metabolism by the indigenous soil microbiota was always accompanied by a significant decrease in the pH of soil solutions, but not necessarily by an increase in the viable count. The feasibility of using electrical conductivity and water-soluble Ca and Mg measurements to detect metabolic activity, either alone or in conjunction with other life detection techniques, is discussed.

Aerobiosis↗
PubMed2001

Inflammation and clearance of Chlamydia trachomatis in enteric and nonenteric mucosae.

Immunization(s) fostering the induction of genital mucosa-targeted immune effectors is the goal of vaccines against sexually transmitted diseases. However, it is uncertain whether vaccine administration should be based on the current assumptions about the common mucosal immune system. We investigated the relationship between mucosal sites of infection, infection-induced inflammation, and immune-mediated bacterial clearance in mice using the epitheliotropic pathogen Chlamydia trachomatis. Chlamydial infection of the conjunctival, pulmonary, or genital mucosae stimulated significant changes in tissue architecture with dramatic up-regulation of the vascular addressin, VCAM, a vigorous mixed-cell inflammatory response with an influx of alpha4beta1+ T cells, and clearance of bacteria within 30 days. Conversely, intestinal mucosa infection was physiologically inapparent, with no change in expression of the local MAdCAM addressin, no VCAM induction, no histologically detectable inflammation, and no tissue pathology. Microbial clearance was complete within 60 days in the small intestine but bacterial titers remained at high levels for at least 8 months in the large intestine. These findings are compatible with the notion that VCAM plays a functional role in recruiting cells to inflammatory foci, and its absence from the intestinal mucosa contributes to immunologic homeostasis at that site. Also, expression of type 1 T cell-mediated immunity to intracellular Chlamydia may exhibit tissue-specific variation, with the rate and possibly the mechanism(s) of clearance differing between enteric and nonenteric mucosae. The implications of these data for the common mucosal immune system and the delivery of vaccines against mucosal pathogens are discussed.

Animals↗
PubMed1978

Relative microbial activity and bacterial concentrations in water and sediment samples taken in the Beaufort Sea.

A total of 91 water, 8 ice, and 50 sediment samples taken from the southwestern Beaufort Sea were analyzed for relative microbial activity, bacterial cell concentrations, and percentage of respiration (mineralization). These samples were taken during three field-study periods (August to September 1975; April 1976; and August 1976). Both the relative microbial activity and the cell concentrations in water and sediment samples were lower during the April (winter) sampling period than in the August-September (summer) studies. The percentage of respiration of labeled glutamic acid was higher in the winter water samples than in the summer samples. The water samples showed higher percentage of respiration values than did sediment samples. The average maximimum potential rate of glutamic acid uptake was as high or higher than those observed in studies made in more temperate waters. Samples of melted sea ice showed levels of relative microbial activity that were about the same as that found in the associated seawater. When 1:1 mixtures of melted ice and seawater were analyzed for altered microbial activity, little effect could be detected.

Alaska↗
PubMed2006

Recognition and signaling by toll-like receptors.

Toll-like receptors (TLRs) are transmembrane proteins that detect invading pathogens by binding conserved, microbially derived molecules and that induce signaling cascades for proinflammatory gene expression. A critical component of the innate immune system, TLRs utilize leucine-rich-repeat motifs for ligand binding and a shared cytoplasmic domain to recruit the adaptors MyD88, TRIF, TIRAP, and/or TRAM for downstream signaling. Despite significant domain conservation, TLRs induce gene programs that lead not only to the robust production of general proinflammatory mediators but also to the production of unique effectors, which provide pathogen-tailored immune responses. Here we review the mechanisms by which TLRs recognize pathogens and induce distinct signaling cascades.

Animals↗
PubMed2006

Origin of mutations under selection: the adaptive mutation controversy.

Growth under selection causes new genotypes to predominate in a population. It is difficult to determine whether selection stimulates formation of new mutations or merely allows faster growth of mutants that arise independent of selection. In the practice of microbial genetics, selection is used to detect and enumerate pre-existing mutants; stringent conditions prevent growth of the parent and allow only the pre-existing mutants to grow. Used in this way, selection detects rare mutations that cause large, easily observable phenotypic changes. In natural populations, selection is imposed on growing cells and can detect the more common mutations that cause small growth improvements. As slightly improved clones expand, they can acquire additional mutational improvements. Selected sequential clonal expansions have huge power to produce new genotypes and have been suggested to underlie tumor progression. We suggest that the adaptive mutation controversy has persisted because the distinction between these two uses of selection has not been appreciated.

Adaptation, Physiological↗
PubMed2005

Lipopolysaccharide heterogeneity: innate host responses to bacterial modification of lipid a structure.

The innate host response system is composed of various mechanisms designed to detect and facilitate host responses to microbial components, such as lipopolysaccharides (LPS). To enable this to occur, innate systems contain multiple pattern recognition receptors (i.e., LBP, CD14, and TLRs), which identify certain features within bacterial LPS that are foreign to the host, as well as essential and uniquely specific for bacteria. Innate host identification of unique bacterial components or patterns, therefore, relies on the inability of bacteria to alter these essential or critical components dramatically. Historically, LPS have been viewed as essential outer-membrane molecules containing both a highly variable outer region (O-segment) as well as a relatively conserved inner region (lipid A). However, over the last decade, new evidence has emerged, revealing that increased natural diversity or heterogeneity within specific components of LPS, such as lipid A-resulting in minor to moderate changes in lipid A structure-can produce dramatic host responses. Therefore, examples of natural lipid A heterogeneity, and the mechanisms that control it, represent a novel approach in which bacteria modulate host responses and may thereby confer specific advantages to certain bacterial species under changing environmental host conditions.

Animals↗
PubMed2005

High-level expression of soluble human beta-defensin-2 fused with green fluorescent protein in Escherichia coli cell-free system.

Human beta-defensin-2 (hBD2), a small cationic peptide, exhibits a broad range of antimicrobial activity and does not acquire any microbial resistance. To produce this uneasily detectable, degradable, and toxic polypeptide efficiently, an alternative approach based on the Escherichia coli cell-free biosynthesis system was proposed. The approach implies that a polypeptide of interest is synthesized as a fusion protein linked to a green fluorescent protein (GFP) through a cleavable spacer. With batch-mode operation, a significant amount of hBD2 fused with GFP (0.25 mg/mL) can be expressed in this cell-free system. The productivity of the fusion protein can be improved up to 1.2 mg/mL by employing a continuous-exchange cell-free system. Furthermore, the GFP moiety provides directly visible and quantitative monitoring of the polypeptide synthesis, and the product is soluble and stable. This work will be helpful in allowing the rapid and visible expression of other similar defensins using an in vitro cell-free system.

Escherichia coli↗
PubMed1996

Madindoline, a novel inhibitor of IL-6 activity from Streptomyces sp. K93-0711. I. Taxonomy, fermentation, isolation and biological activities.

Selective growth inhibition against IL-6-dependent cells was detected in fermentation extracts of a microbial strain, K93-0711, which was characterized as Streptomyces species. Active metabolite, termed madindoline A and B, were isolated, and the structure was determined to be 3a-hydroxy-indoline with diketocyclopentene at the N position. Madindoline A and B displayed dose-dependent inhibition of MH60 cells, an IL-6-dependent cell line, in presence of 0.1 U/ml IL-6. The IC50 for madindoline A and B against this cell line was 8 microM and 30 microM, respectively. These compounds did not inhibit the growth of cell lines which are not IL-6 dependent and the growth inhibition of the MH60 cell line was reversed by addition of excess, 0.4 U/ml, of IL-6 to the culture media. These compounds did not show any antimicrobial activity at a concentration of 1,000 micrograms/ml.

Bridged Bicyclo Compounds, Heterocyclic↗
PubMed1978

[Simple qualitative methods of detecting beta-lactamase-producing strains of microorganisms].

Modifications of two microbiological qualitative methods for detection of beta-lactamase-producing microbial strains are described. The methods are based on the principle of Goths. One of the methods named "contact" significantly differs from the prototype and is more simple, convenient, easily reproducible and does not require the use of special equipment and special training of the staff.

Bacteria↗
PubMedDate not supplied

Adenosine triphosphate bioluminescence analysis for rapid screening of microbial contamination in non-sterile pharmaceutical samples.

An Adenosine Triphosphate (ATP) bioluminescence system was compared and validated against standard methods for rapid microbiological monitoring of several non-sterile pharmaceutical formulations such as creams, tablets, and capsules. Results obtained using 1%, 2.5%, and 10% of product suspensions indicated that most samples that did not contain non-microbial ATP neither inhibited the bioluminescence reaction nor did something else. Ten percent product suspensions were inoculated with different concentrations of Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, Candida albicans, and Aspergillus niger. Samples were incubated for 24-120 h at 35 degrees C with shaking. Results indicated a strong inhibitory effect of microbial growth, as no microorganisms were detected by using the ATP bioluminescence assay. However, when 1% and 2.5% product suspensions were spiked with the same microorganisms, positive detection was confirmed. After incubation, all microorganisms were detected by the bioluminescence system within 24-72 h. All positive samples were confirmed by using standard plating media. However, to optimize detection of all microorganisms, different enrichment media were developed.

Adenosine Triphosphate↗
PubMed2005

Sequence-specific cleavage of 16S rRNA for rapid and quantitative detection of particular groups of anaerobes in bioreactors.

We developed a rapid and simple method for rRNA-based quantitative detection of a specific group of microorganisms in complex ecosystems. The method relies on the sequence-specific scission of 16S rRNA with ribonuclease H (RNase H) and oligonucleotides that specifically hybridize with targeted rRNA molecules. RNAs from a complex community were first mixed with an oligonucleotide and were subsequently digested with RNase H to achieve sequence-dependent rRNA cleavage at the hybridization site. For the quantitative detection of targeted rRNAs, the resulting RNA fragment patterns were analyzed by gel-electrophoresis, which separated and quantified cleaved and intact rRNA fragments. This method enabled the quantitative detection of microbes in a complex microbial community by a relatively simple and fast experimental procedure. We then applied the cleavage method to actual anaerobic microbial communities such as digested sewage sludge and UASB sludges. The results demonstrated that the present method was fully applicable to anaerobic digestor ecosystems containing complex anaerobic microorganisms.

Bioreactors↗
PubMed2006

[Genotype distribution of extended-spectrum beta-lactamases and AmpC beta-lactamases produced in Escherichia coli isolated from men with urinary infection].

OBJECTIVE: To study the genotype distribution of extended-spectrum p-lactamases (ESBLs) and AmpC p-lactamases produced in E. coli isolated from men with urinary infection in Nanjing. METHODS: Organisms of clinical infection were identified by automatic microbial system (Vitek-32). ESBLs were detected by disk diffusion confirmatory test, and ESBLs and AmpC p-lactamases by three-dimensional extract test (TDET) , the presence of plasmid-mediated ESBLs and ampC genes determined by PCR, and conjugal transfer assays of the ampC resistance determinants carried out by a broth mating procedure. RESULTS: ESBLs were produced in 24. 6% (46/187) of the E. coli and the 46 E. coli isolates showed p-lactamase activity in TDET, 3 positive for both ESBLs and AmpC beta-lactamases and 43 for ESBLs only. Forty-four of the 46 isolates were shown by PCR to contain at least one of the genes blaTEM, blaOXA, bla(CTX-M), but no blaSHA. AmpC specific amplication products were observed in 3 of the 46 isolates, of which 2 were of CIT type, and 1 of DHA type. All of the 3 transconjugants transferred the plasmids harbouring ampC genes to recipients. CONCLUSION: CTX-M is the most common genotype in plasmid-mediated ESBLs produced by E. coli isolated from men with urinary infection in Nanjing. Present findings indicate that AmpC-producing E. coli are present in this hospital, and ampC-encoding plasmids are transferable.

Bacterial Proteins↗
PubMed1990

[Ways of improving the effectiveness of bacteriological diagnosis and treatment of nonspecific lung diseases].

Isolation, identification and drug sensitivity assay of microorganisms from pathological materials of 177 patients with nonspecific diseases of the lungs, mainly pneumonia, were performed on blood and selective "chocolate" agars by using Baktofok-MK, a new dry nutrient basis developed by the authors. Blood and "chocolate" agars based on the Hottinger's hydrolysate were used as the control media. It was shown that with the quantitative procedure for inoculating the pathological material, the experimental media based on Baktofok-MK were much more sensitive to growth properties that the control media. That made it possible to detect larger numbers of etiologically important microbial species on the blood agar and to isolate clinical strains of Hemophilus spp. from a larger number of specimens on the "chocolate" agar.

Agar↗
PubMed1989

[Bacterial infection and rapid laboratory microbial methods].

The key to treatment of bacterial infectious diseases is always to quickly identify the causative organism and understand its resistance to drugs. Recent progress in microbial laboratory methods has permitted rapid detection and identification of pathogenic organisms. Rapid test methods are classified into culture and non-culture methods. Non-culture methods are based mainly on immunoassay for detection of antigen or antibody of pathogenic organisms, but also include the DNA probe method and the RNA probe method. Immunoassay is achieved by fluorescent antibody techniques, agglutination and ELISA. On a related note, monoclonal antibodies have been developed with steady progress. For culture methods, commercial bacterial identification kits and automated instruments, all of which permit quicker identification than with conventional methods, are now used in a large number of laboratories. Quick identification with these automated instruments is possible owing to optical determination of drug resistance. Some automated instruments are capable of rapidly detecting bacteria in the sample. For example, Bactec, used for quick diagnosis of bacteremia, measures CO2 produced by bacteria in the culture bottle during the metabolic process, permitting early detection of bacterial proliferation. Other methods are available in which ATP produced by bacteria is measured on the basis of bioluminescence or chemiluminescence.

Bacteria↗