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PubMed1995

Effects of free and liposome-encapsulated antibiotics on adherence of Pseudomonas aeruginosa to collagen type I.

The adherence of 27 clinical Pseudomonas aeruginosa strains to collagen type I was investigated by using a solid-phase assay. The influence of free antibiotics (amikacin, gentamicin, piperacillin, bacitracin, and polymyxin B) and liposome-entrapped antibiotics (amikacin and polymyxin B) on bacterial attachment to collagen type I was examined. The greatest inhibitory effect was shown for free and liposomal amikacin, which decreased the attachment of 74 and 100% of tested strains, respectively. The mean percent attachment (+/- standard deviation) in the presence of free amikacin was 65.7% (+/- 12.0%) as measured by solid-phase assay. In the presence of liposomal amikacin, the attachment ranged from 17.3% (+/- 6.0%) to 42.1% (+/- 9.4%), depending on the antibiotic solvent. In contrast, polymyxin B, even at a subinhibitory concentration, enhanced attachment of all P. aeruginosa isolates to collagen. Liposomal polymyxin B displayed a protective effect only when the encapsulated drug was of a low concentration. Application of liposome-encapsulated amikacin may be advantageous in injured tissues in which extracellular matrix structures become exposed.

Amikacin↗
PubMed1987

[3H]QNB binding and contraction of rabbit colonic smooth muscle cells.

We used radioligand binding and studies of cell contraction to characterize muscarinic receptors on dispersed smooth muscle cells from rabbit proximal and distal colon. Cells obtained after serial incubations in collagenase were used to measure binding of tritiated quinuclidinyl benzilate [( 3H]QNB). AT 37 degrees C, specific [3H]QNB binding was saturable and linearly related to cell number. Nonlinear regression analysis was used to determine the affinity of [3H]QNB for its receptor. In the distal colon the Kd was 60 pM, and the mean number of receptors was 1.2 X 10(6)/cell. Compared with cells from the distal colon, cells from the proximal colon had a lower affinity (Kd = 337 pM) but similar numbers of receptors. The concentrations of the antagonists atropine, secovorine, and pirenzepine, which were required for inhibition of 50% [3H]QNB binding (IC50), were 8, 5, and 870 nM, respectively, suggesting that the receptors are of the M2-muscarinic subclass. Hill coefficients for these agents were 1.1, 0.9, and 1.1, suggesting binding to a single receptor. The IC50 for the muscarinic agonists bethanechol and oxotremorine were 80 and 0.57 microM, respectively. Hill coefficients were 0.67 for both, suggesting more complex interactions involving receptors of different affinities. In studies of cell contraction, bethanechol stimulated a dose-dependent decrease in cell length with half the maximal contraction occurring at 100 pM. These results suggest that 1) contraction is mediated by binding of bethanechol to M2-muscarinic receptors and that 2) there are a large number of spare receptors in colonic smooth muscle.

Animals↗
PubMed2006

Decomposers (Lumbricidae, Collembola) affect plant performance in model grasslands of different diversity.

Decomposer invertebrates influence soil structure and nutrient mineralization as well as the activity and composition of the microbial community in soil and therefore likely affect plant performance and plant competition. We established model grassland communities in a greenhouse to study the interrelationship between two different functional groups of decomposer invertebrates, Lumbricidae and Collembola, and their effect on plant performance and plant nitrogen uptake in a plant diversity gradient. Common plant species of Central European Arrhenatherion grasslands were transplanted into microcosms with numbers of plant species varying from one to eight and plant functional groups varying from one to four. Separate and combined treatments with earthworms and collembolans were set up. Microcosms contained 15N labeled litter to track N fluxes into plant shoots. Presence of decomposers strongly increased total plant and plant shoot biomass. Root biomass decreased in the presence of collembolans and even more in the presence of earthworms. However, it increased when both animal groups were present. Also, presence of decomposers increased total N concentration and 15N enrichment of grasses, legumes, and small herbs. Small herbs were at a maximum in the combined treatment with earthworms and collembolans. The impact of earthworms and collembolans on plant performance strongly varied with plant functional group identity and plant species diversity and was modified when both decomposers were present. Both decomposer groups generally increased aboveground plant productivity through effects on litter decomposition and nutrient mineralization leading to an increased plant nutrient acquisition. The non-uniform effects of earthworms and collembolans suggest that functional diversity of soil decomposer animals matters and that the interactions between soil animal functional groups affect the structure of plant communities.

Animals↗
PubMed1977

Uptake of 2'-amino-2'-deoxyguanosine by Escherichia coli and its competition by guanosine.

The antibacterial activity of a new nucleoside antibiotic, 2'-amino-2'-deoxyguanosine (2AG), is reversed by guanosine and other purine nucleosides. 2AG is apparently taken up by E. coli by a mechanism different from that of guanosine; guanosine inhibits this uptake non-competitively. Insensitive E. coli strains and the resistant mutant obtained from the sensitive strain also took up 2AG.

Binding, Competitive↗
PubMedDate not supplied

Improved DNA probe detection of Listeria monocytogenes in enrichment culture after physical-chemical fractionation.

Bacterial detection in foods by nucleic acid probes is limited by microflora competition during selective enrichment. Probe target concentration by extraction and fractionation of enrichments may diminish this limitation. The 1-h AccuProbe chemiluminescent culture identification test for Listeria monocytogenes was used as a model. Its high detection threshold provides a stringent challenge for evaluating enrichment work-up protocols. Detection of L. monocytogenes, at 1-4 colony-forming units/g food, was not consistently possible in 48 h enrichment cultures using AccuProbe. Concentration by cell sedimentation was occasionally helpful but the volume of co-sedimented food limited concentration to about 10-fold. To improve concentration, enrichment sediments were sonicated or enzymatically lysed to release the probe's target, r-RNA. The RNA was separated from non-RNA material by extraction with phenol and precipitation with ethanol. Enrichments (250 mL) were concentrated 2500-fold, and the limitation was food RNA volume. A strongly competitive Enterococcus faecium food isolate was used to demonstrate the effect of artificial competition on the kit's ability to detect L. monocytogenes in enrichments. High competitor concentrations repressed the level of the target below the detection threshold, but concentration of r-RNA enabled detection of L. monocytogenes. The effectiveness of this enrichment sample work-up was demonstrated with naturally contaminated hummus.

Bacteriological Techniques↗
PubMed1998

Implications of resource-ratio theory for oral microbial ecology.

The ability to compete for the limited nutrients available to the microorganisms of dental plaque is a strong ecological determinant of the structure of the subgingival ecosystem. This paper introduces a new concept from the field of ecology, resource-ratio theory, and applies it to the dynamics of microbial dental plaque with emphasis on the putative periodontal pathogens. Resource-ratio theory is a mechanistic theory of resource competition that utilizes pairs of growth-limiting nutrients in a stoichiometric fashion to predict zones of competitive dominance, exclusion, and coexistence for organisms competing for these resources. Once these resource pairs are identified for plaque organisms, resource-ratio theory may provide predictions of changes in the microbial community structure of plaque based on directional changes in their resource supply ratios.

Bacteria↗
PubMed1982

Sorption of cadmium by microorganisms in competition with other soil constituents.

The fate of cadmium in soil is influenced to a great extent by microbial activity. Microorganisms were compared with abiotic soil components for their ability to sorb Cd from a liquid medium. When the same amount (on a dry weight basis) of bacterial cells (Serratia marcescens and Paracoccus sp.), clay (montmorillonite), or sand was separately incubated in 0.05 M phosphate buffer, pH 7.2, containing 10 ppm of Cd (10 mug/ml), bacterial cells removed the largest quantity of Cd. Dead cells sorbed much more Cd from the medium than live cells. A comparative study of Cd removal from the medium by seven soil bacteria and four fungi did not indicate appreciable differences. With increasing microbial biomass, the relative efficiency of 0.1 M NaOH as an extractant of sorbed Cd increased, whereas the extraction efficiency of 0.005 M DTPA (diethylenetriaminepentaacetic acid) decreased. It appeared that NaOH and DTPA extracted different chemical forms of Cd. This assumption was supported by vastly different correlation coefficients in the relative amount of Cd extracted by the two solvents.

Journal Article↗
PubMed2026

Soil erosion and landscape elevation as unnoticed determinants of environmental antibiotic resistance distribution.

Climate change is reshaping the global antibiotic resistance gene (ARG) landscape through geomorphological processes that remain largely overlooked in the One Health framework. This critical review synthesises evidence on how soil erosion and landscape elevation gradients redistribute, select for, and disseminate ARGs across terrestrial and aquatic ecosystems. Erosion physically removes and transports ARG-bearing microbes, depletes nutrients, and co-selects for resistance via heavy metal exposure and horizontal gene transfer, creating source-sink dynamics that connect eroding hillslopes to downstream water bodies and food systems. Elevation gradients impose abiotic stressors-declining temperature, elevated UV radiation, and shifting pH-that drive microbial community reassembly through environmental selection and dispersal limitation, with emerging evidence linking bacterial competition at high altitude to enhanced multidrug efflux and resistome complexity. The review identifies critical knowledge gaps, including unquantified ARG mass fluxes across erosion-deposition gradients, unresolved dispersal-versus-selection mechanisms along elevation transects, and the absence of integrated One Health surveillance linking environmental ARG reservoirs to clinical outcomes. A synthesis of global case studies illustrates how these processes converge across diverse landscapes. The review concludes with a mechanistic research agenda-including reciprocal transplant experiments, landscape connectivity modelling, and cross-sectoral surveillance-needed to translate these emerging drivers into actionable climate-AMR mitigation policy.

Drug Resistance, Microbial↗
PubMed2002

Fitness effects of fixed beneficial mutations in microbial populations.

Beneficial mutations are intuitively relevant to understanding adaptation, yet not all beneficial mutations are of consequence to the long-term evolutionary outcome of adaptation. Many beneficial mutations-mostly those of small effect-are lost due either to (1) genetic drift or to (2) competition among clones carrying different beneficial mutations, a phenomenon called the "Hill-Robertson effect" for sexual populations and "clonal interference" for asexual populations. Competition among clones becomes more prevalent with increasing genetic linkage and increasing population size, and it is thus generally characteristic of microbial populations. Together, these two phenomena suggest that only those beneficial mutations of large fitness effect should achieve fixation, despite the fact that most beneficial mutations produced are predicted to have very small fitness effects. Here, we confirm this prediction-both empirically and theoretically-by showing that fitness effects of fixed beneficial mutations follow a distribution whose mode is positive.

Adaptation, Physiological↗
PubMed2003

Importance of Gram-positive naphthalene-degrading bacteria in oil-contaminated tropical marine sediments.

AIMS: The aim of this study was to isolate, characterize and evaluate the importance of naphthalene-degrading bacterial strains from oil-contaminated tropical marine sediments. METHODS AND RESULTS: Three Gram-positive naphthalene-degrading bacteria were isolated from oil-contaminated tropical intertidal marine sediments by direct isolation or enrichment using naphthalene as the sole source of carbon and energy. Bacillus naphthovorans strain MN-003 can also grow on benzene, toluene, xylene and diesel fuel while Micrococcus sp. str. MN-006 can also grow on benzene. Staphylococcus sp. str. MN-005 can only degrade naphthalene and was not able to use the other aromatic hydrocarbons tested. Strain MN-003 possessed the highest maximal specific growth rate with naphthalene as sole carbon source. An enrichment culture fed with naphthalene as sole carbon source exhibited a significant increase in the relative abundances of the three isolates after 21 days of incubation. The three isolates constituted greater than 69% of the culturable naphthalene-degrading microbial community. Strain MN-003 outcompeted and dominated the other two isolates in competition studies involving batch cultures inoculated with equal cell densities of the three isolates and incubated with between 1 and 10 mg l-1 of naphthalene. CONCLUSIONS: Three Gram-positive naphthalene-degrading bacteria were successfully isolated from oil-contaminated tropical marine sediments. Gram-positive bacteria might play an important role in naphthalene degradation in the highly variable environment of oil-contaminated tropical intertidal marine sediments. Among the three isolates, strain MN-003 has the highest maximal specific growth rate when grown on naphthalene, and outgrew the other two isolates in competition experiments. SIGNIFICANCE AND IMPACT OF THE STUDY: This research will aid in the development of bioremediation schemes for oil-contaminated marine environments. Strain MN-003 could potentially be exploited in such schemes.

Bacillus↗
PubMed1983

Microbial activity at the sediment-water interface in halifax harbor, Canada.

The sediment-water interface in Halifax Harbor supports a microbial population of 6.95 x 10 cells per g (dry weight). As determined by the standard technique of suspending subsamples in filtered seawater, the uptake of added glutamic acid by this population is 113.5 ng g (dry weight) h. An alternate technique was developed to measure the heterotrophic activity of the interface over longer periods of time, using undisturbed cores with the sediment-water interface intact. Under these conditions, the microbes in the water column and the interface increased exponentially in number, with mean doubling times of 9.6 and 4.5 days, respectively. The uptake of glutamic acid by the microbial population of the interface was determined to be 12.7 ng g (dry weight) h, almost an order of magnitude less than the uptake determined by the previous method. This indicates that substrate diffusion and competition for substrate by the microbes in the water column are important factors when considering the heterotrophic activity of the sediment microbial population. After 48 h of incubation, uptake and respiration ceased, probably due to the exhaustion of labeled substrate. Additional substrate added after 48 h of incubation was taken up at a rate similar to that measured after the first addition. It appears that the microbial population of the interface is able to respond quickly and repeatedly to relatively large nutrient additions. After 10 days of incubation, the number of "viable" cells as determined by autoradiography was much smaller than the increase in numbers as determined by direct counts. Apparently a large part of the viable population is unaffected by nutrient addition.

Journal Article↗
PubMed2006

A microbial diagnostic microarray technique for the sensitive detection and identification of pathogenic bacteria in a background of nonpathogens.

A major challenge in microbial diagnostics is the parallel detection and identification of low-bundance pathogens within a complex microbial community. In addition, a high specificity providing robust, reliable identification at least at the species level is required. A microbial diagnostic microarray approach, using single nucleotide extension labeling with gyrB as the marker gene, was developed. We present a novel concept applying competitive oligonucleotide probes to improve the specificity of the assay. Our approach enabled the sensitive and specific detection of a broad range of pathogenic bacteria. The approach was tested with a set of 35 oligonucleotide probes targeting Escherichia coli, Shigella spp., Salmonella spp., Aeromonas hydrophila, Vibrio cholerae, Mycobacterium avium, Mycobacterium tuberculosis, Helicobacter pylori, Proteus mirabilis, Yersinia enterocolitica, and Campylobacter jejuni. The introduction of competitive oligonucleotides in the labeling reaction successfully suppressed cross-reaction by closely related sequences, significantly improving the performance of the assay. Environmental applicability was tested with environmental and veterinary samples harboring complex microbial communities. Detection sensitivity in the range of 0.1% has been demonstrated, far below the 5% detection limit of traditional microbial diagnostic microarrays.

Bacteria↗
PubMed2002

A surface plasmon resonance analysis of the interaction between the antibiotic moenomycin A and penicillin-binding protein 1b.

The antibiotic moenomycin A inhibits the biosynthesis of peptidoglycan, the main structural polymer of the bacterial cell wall. The inhibition is based on a reversible binding of the antibiotic to one of the substrate binding sites in enzymes such as penicillin-binding protein (PBP) 1b. A novel assay based on surface plasmon resonance (SPR) has been established that can be used to investigate selective binding of the moenomycin sugar moiety and other transglycosylase inhibitors to this enzyme. Suitable ligands were prepared from moenomycin A and coupled to SPR sensor surfaces. Moenomycin analogues with structural variations were used to perform competitive SPR experiments with PBP 1b. The SPR results confirm for the first time that the trisaccharide fragment of moenomycin A (C-E-F-G-H-I) is the minimal structure that possesses all moieties sufficient for biological activity and for affinity towards PBP 1b. The method seems to be appropriate for use in screens for transglycosylase inhibitors that bind to the moenomycin-binding site of the enzyme.

Bacterial Proteins↗
PubMed1990

The collagenase gene promoter contains a TPA and oncogene-responsive unit encompassing the PEA3 and AP-1 binding sites.

PEA3 is a transcription factor which binds to the polyoma virus enhancer and whose activity is regulated by the expression of a number of oncogenes. We show here that PEA3 also binds specifically to the collagenase and fos cellular promoters. On the collagenase promoter, PEA3 acts synergistically with AP-1 to achieve maximum levels of transcription activation by 12-O-tetradecanoylphorbol-13-acetate (TPA), and non-nuclear oncoproteins, thereby defining a TPA- and oncogene-responsive unit (TORU). From a comparative study of the collagenase TORU and the analogous polyoma virus TORU, we conclude that both the binding affinity of the PEA3 motif and the spacing between PEA3 and AP-1 modulate transcription activation induced by oncogene expression.

Base Sequence↗
PubMed1999

Resistance responses of microorganisms in food environments.

Food borne microorganisms display a broad spectrum of resistance responses to naturally occurring and intentionally added antimicrobial agents. Resistance may be conferred by innate structural features of the bacterial strain such as an impermeable outer membrane or a mechanism for antibiotic-inactivation. Bacteria previously susceptible to an antimicrobial compound can acquire resistance through mutation or through genetic transfer processes such as transformation, transduction, and conjugation. Resistance can also be conferred by biofilm formation on food processing surfaces as an adaptive response to protect colonies from cleaning and sanitation. Resistant pathogens are a global problem, facilitated by international trade of raw and processed foods. Cross resistance between clinical and nonclinical antimicrobials can exist and is of concern. The development of resistant foodborne pathogens has been attributed to increased antibiotic use in hospitals, outpatient facilities, and veterinary applications. Resistant microorganisms can also develop as a result of physical processes used in food preservation, such as acid treatments and irradiation processes. Strategies to effectively counter resistance development include: changing current practices of antibiotic usage, developing new antibiotics, applying hurdle preservation approaches, preventing bacterial adhesion, and utilizing competitive exclusion. This paper presents an overview of problems arising from the development of microbial resistance, and explores possible solutions for detecting and defeating the adaptive changes of microorganisms.

Drug Resistance, Microbial↗