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In vitro activity of fleroxacin in combination with other antimicrobial agents.

The trifluoroquinolone fleroxacin inhibits the majority of Enterobacteriaceae at concentrations < or = 1 micrograms/mL and most Pseudomonas aeruginosa and staphylococci at < or = 2 micrograms/mL. The purpose of this study was to determine the effect of the combination of fleroxacin with other antimicrobial agents. Previous studies that used checkerboard assay, fixed concentrations, and killing curves were reviewed, and these methods were used to evaluate the combination of fleroxacin and agents that had not been previously studied. The combination of fleroxacin with such aminoglycosides as gentamicin, amikacin, and tobramycin is indifferent against most Enterobacteriaceae, as is the combination of fleroxacin with penicillins, cephalosporins, rifampin, clindamycin, and metronidazole. Combinations of fleroxacin with penicillins, cephalosporins, imipenem, aminoglycosides, clindamycin, metronidazole, and rifampin are indifferent against P. aeruginosa. Fosfomycin and fleroxacin acted synergistically against P. aeruginosa. Against staphylococci, combinations of fleroxacin with oxacillin, rifampin, or fosfomycin had synergistic or additive effects, whereas combinations of fleroxacin with vancomycin, gentamicin, or metronidazole have shown indifference. No synergy or antagonism has been found for combinations of fleroxacin with penicillin, vancomycin, erythromycin, clindamycin, or rifampin against streptococci or enterococci. The combination of fleroxacin and metronidazole has proved synergistic against various Bacteroides species. In general, combinations of fleroxacin with other antimicrobial agents display indifference and rarely synergy. Thus, fleroxacin can be combined with other antibiotics to enlarge the spectrum of activity.

Anti-Bacterial Agents↗

Effect of pesticides on growth of rhizobia and their host plants during symbiosis.

Effect of various pesticides (insecticides, fungicides and herbicides) has been studied on growth and efficiency of symbiotic properties of 3 fast growing Rhizobium sp. under green house conditions. The results revealed adverse effects on plant growth and nitrogen fixing capacity as measured by dry weight and total nitrogen content of plants infected with pesticide treated Rhizobium. Of the pesticides tested, herbicides were found to be more effective on the above parameters than the insecticides and fungicides.

Fabaceae↗

Induction of in vitro human macrophage anti-Mycobacterium tuberculosis activity: requirement for IFN-gamma and primed lymphocytes.

Mycobacterium tuberculosis (Mtb) is the world's leading infectious cause of mortality. Despite the overwhelming data supporting the critical role of cellular immunity, little is known of the early microbial and immune cell interactions and whether human macrophages can be activated to express anti-Mtb activity. We report the reconstitution of an in vitro system whereby human macrophages express anti-Mtb activity only in coculture with PBL and with IFN-gamma. Omission of IFN-gamma in the cocultures or Mtb lysate/IFN-gamma-primed lymphocytes was associated with high growth of Mtb, high IL-10 and IL-12 p40, nearly undetectable IL-12 p70 levels, and the highest percentages of CD4 and CD8 T cells. In contrast, IFN-gamma treatment of cocultures containing Mtb lysate/IFN-gamma-primed PBL reduced bacilli count by approximately 2.5 log, decreased the production of IL-10 by 5.7-fold, increased IL-12 p70 by approximately 50-fold, and reduced the percentages of CD4 and CD8 T cells. Activation of anti-Mtb activity was time and dose dependent. At 2000 U/ml of IFN-gamma, bactericidal activity was achieved (10-fold reduction from initial inoculum). Anti-Mtb activity against several strains of M. tuberculosis (H37Ra and H37Rv, and C, a clinical isolate) was observed and was associated with expression of inducible nitric oxide synthase. These data suggest that induction of human macrophage anti-Mtb activity required dual signaling from PBL and IFN-gamma. Thus, the development of an in vitro human system may greatly facilitate studies to delineate immune cells, cytokines, and effector functions/genes critical in controlling Mtb. Defining the mechanisms may also provide novel treatment strategies for tuberculosis.

Antigen Presentation↗

Antibiotic-mediated release of endotoxin and the pathogenesis of gram-negative sepsis.

Since the earliest days of antibiotic chemotherapy to treat infection with Gram-negative microbes, investigators have recognized that such treatments may result in the release of microbial constituents that might, in turn, exacerbate the pathophysiological manifestations of disease. Both in vitro studies and in vivo animal experiments have over the years provided evidence in support of this concept; however, the actual clinical importance of this phenomenon to patients with Gram-negative sepsis is unclear. Recently published reports from a number of laboratories have shown that cell wall-active antibiotics that differ in their fundamental mechanisms of action in disrupting microbial growth (via selective interactions with various penicillin binding proteins) also differ in their relative ability to induce the release of biologically active endotoxin both in vitro and in vivo. Further, quantitative differences in total endotoxin release correlate well with antibiotic-initiated morphological changes in the microbe. Of potential significance is the finding that these differences are also reflected in differential production of cytokines from endotoxin-stimulated mononuclear phagocytes and other host target cells, including 11-6 and TNF. Since these immunologic hormones have been strongly implicated as contributing factors to the pathogenesis of Gram-negative sepsis, interest in the potential use of this chemotherapeutic approach as a means of controlling the host immunopathologic response has increased. Carefully controlled clinical trials in which different antibiotic treatments are correlated with production of cytokines will be of significant potential value in evaluating the actual significance of this phenomenon in the Gram-negative septic patient.

Animals↗

Plaque mineralisation in vitro.

Dental calculus is plaque mineralised by deposition of calcium and phosphate resulting from interactions between the oral microbial plaque flora and components of oral fluids. An artificial-mouth microcosm dental plaque culture system has been developed to study aspects of plaque mineralisation, including pH control. Five plaques were grown from saliva under simulated oral conditions in a mucin-containing medium, and sucrose was applied to mimic meals. The plaques were mineralised with a urea-based, calcium-phosphate-monofluorophosphate-urea (CPMU) mineralising solution. Alkaline pH oscillations were generated by the plaques in response to CPMU applications, and an acidic oscillation followed sucrose applications. Plaque mineralisation by the CPMU procedure was almost totally dependent on the urea present in the mineralising solution, but total mineralisation also increased as the resting pH increased as a result of urea in the medium. Following four CPMU applications with a sucrose application every 12 hours improved plaque viability and mineralisation. The plaque mineral formed resembled a carbonated hydroxyapatite; other potential calcium phosphate minerals were undetectable except for calcium carbonate. A wide range of mineral deposition patterns in plaque were seen by electron microscopy.

Acids↗

How Drosophila combats microbial infection: a model to study innate immunity and host-pathogen interactions.

During the past year, dramatic progress has been achieved in our understanding of Drosophila immune reactions. The completion of the Drosophila genome sequencing project, microarray analysis and the use of genetic screens have led to the identification of several new genes required to combat microbial infection, filling in some important gaps in the understanding of innate immunity. At the same time, this insect was used as a model for the study of host-pathogen interactions. The recent major advances on the mechanisms by which this insect defends itself against intrusion of pathogens are discussed in this review.

Animals↗

Lipopolysaccharide interaction with cell surface Toll-like receptor 4-MD-2: higher affinity than that with MD-2 or CD14.

Toll-like receptors (TLRs) are innate recognition molecules for microbial products, but their direct interactions with corresponding ligands remain unclarified. LPS, a membrane constituent of gram-negative bacteria, is the best-studied TLR ligand and is recognized by TLR4 and MD-2, a molecule associated with the extracellular domain of TLR4. Although TLR4-MD-2 recognizes LPS, little is known about the physical interaction between LPS and TLR4-MD-2. Here, we demonstrate cell surface LPS-TLR4-MD-2 complexes. CD14 greatly enhances the formation of LPS-TLR4-MD-2 complexes, but is not coprecipitated with LPS-TLR4-MD-2 complexes, suggesting a role for CD14 in LPS loading onto TLR4-MD-2 but not in the interaction itself between LPS and TLR4-MD-2. A tentative dissociation constant (Kd) for LPS-TLR4-MD-2 complexes was approximately 3 nM, which is approximately 10-20 times lower than the reported Kd for LPS-MD-2 or LPS-CD14. The presence of detergent disrupts LPS interaction with CD14 but not with TLR4-MD-2. E5531, a lipid A antagonist developed for therapeutic intervention of endotoxin shock, blocks LPS interaction with TLR4-MD-2 at a concentration 100 times lower than that required for blocking LPS interaction with CD14. These results reveal direct LPS interaction with cell surface TLR4-MD-2 that is distinct from that with MD-2 or CD14.

Animals↗

Effect of pyocyanin on a crude-oil-degrading microbial community.

Pseudomonas aeruginosa is an n-alkane degrader that is frequently isolated from petroleum-contaminated sites and produces factors that enhance its competitiveness and survival in many environments. In this study, one such factor, pyocyanin, has been detected in an oil-degrading culture containing P. aeruginosa and is a redox-active compound capable of inhibiting microbial growth. To examine the effects of pyocyanin further, an oil-degrading culture was grown with and without 9.5 microM pyocyanin and microbial community structure and oil degradation were monitored for 50 days. Denaturing gradient gel electrophoresis (DGGE) analysis of cultures revealed a decrease in the microbial community diversity in the pyocyanin-amended cultures compared to that of the unamended cultures. Two members of the microbial community in pure culture exhibited intermediate and high sensitivities to pyocyanin corresponding to intermediate and low levels of activity for the antioxidant enzymes catalase and superoxide dismutase, respectively. Another member of the community that remained constant in the DGGE gels over the 50-day culture incubation period exhibited no sensitivity to pyocyanin, corresponding to a high level of catalase and superoxide dismutase when examined in pure culture. Pyocyanin also affected the overall degradation of the crude oil. At 50 days, the culture without pyocyanin had decreased polycyclic aromatic hydrocarbons compared to the pyocyanin-amended culture, with a specific reduction in the degradation of dibenzothiophenes, naphthalenes, and C(29) and C(30) hopanes. This study demonstrated that pyocyanin influenced the diversity of the microbial community and suggests the importance of understanding how interspecies interactions influence the degradation capability of a microbial community.

Bacteria↗

Problems and priorities for controlling opportunistic pathogens with new antimicrobial strategies; an overview of current literature.

An International Study Group on New Antimicrobial Strategies (ISGNAS) has been formed in response to the recognition that development of microbial resistance to antibiotics is becoming a serious, world-wide problem. The group met in 1993 for the first time to discuss the feasibility of developing rational alternatives to the use of antibiotics and prepared, as a result, a comprehensive overview of normal (physiological) mechanisms involved in the control of potentially pathogenic (oppotunistic) microorganisms. One objective of ISGNAS is to understand the conditions which allow opportunistic microbes present among the symbionts to cause an infection. There is a need for more coherent information concerning the habitat, growth requirements and host and pathogen properties which allow opportunistic pathogens to cause life-threatening infections. In particular, information is urgently being sought to understand the complexity of the interactions between the vast number of microbial species, and the interactions between the microbes and their host. Another goal is to inspire and enable basic and clinical research that will lead to the development of new therapies for regulating colonization, translocation and infection by opportunistic micro-organisms in patients during periods of decreased resistance. With a sufficient amount of knowledge of how healthy individuals keep opportunistic micro-organisms under control, it may become feasible for physicians to maintain host resistance and inter-microbial factors involved in the containment of opportunistic microbes. Therapies aimed at boostering natural resistance mechanisms will be of critical importance to individuals whose resistance has been compromised as a result of another clinical condition.

Adjuvants, Immunologic↗

Propionibacterium acnes-metabolites inhibit experimental lung metastasis of murine sarcoma L-1 in BALB/c-mice.

Adhesive interactions between tumor cells and host tissue occur at several stages of metastasis. Such interactions might be inhibited by microbial metabolites resembling the binding regions of matrix molecules. Certain metabolite sequences including Gly, Asp, Arg, and Ser (GAAS) proved to be critical for cell interactions, e.g. with fibronectin. In vitro, the rosette formation of murine pulmonary cells and sarcoma L-1 cells decreased significantly in the presence of Propionibacterium acnes-metabolites rich in GAAS. In vivo, coinjection of Propionibacterium acnes-metabolites and sarcoma L-1 cells significantly inhibited the formation of lung colonies in BALB/c mice. The inhibition of lung colonization by these metabolites appeared to be noncytotoxic and obviously did not result from impairment of cellular tumorigenicity.

Amino Acid Sequence↗

Effect of alpha-stable sorptive waiting times on microbial transport in microflow cells.

The interaction of bacteria in the fluid phase with pore walls of a porous material involves a wide range of effective reaction times which obey a diversity of substrate-bacteria adhesion conditions, and adhesive mechanisms. For a transported species, this heterogeneity in sorption conditions occurs both in time and space. Modern experimental methods allow one to measure adhesive reaction times of individual bacteria. This detailed information may be incorporated into nonequilibrium transport-sorption models that capture the heterogeneity in reaction times caused by varying chemical conditions. We have carried out particle (Brownian dynamic) simulations of adhesive, self-motile bacteria convected between two infinite plates as a model for a microflow cell. The adhesive heterogeneity is included by introducing adhesive reaction time (understood as time spent at a solid boundary once the particle collides against it) as a random variable that can be infinite (irreversible sorption) or vary over a wide range of values. This is made possible by treating this reaction time random variable as having an alpha-stable probability distribution whose properties (e.g., infinite moments and long tails) are distinctive from the standard exponential distribution commonly used to model reversible sorption. In addition, the alpha-stable distribution is renormalizable and hence upscalable to complex porous media. Simulations are performed in a pressure-driven microflow cell. Bacteria motility (driven by an effective Brownian force) acts as a dispersive component in the convective field. Upon collision with the pore wall, bacteria attachment or detachment occurs. The time bacteria spend at the wall varies over a wide range of time scales. This model has the advantage of being parsimonious, that is, involving very few parameters to model complex irreversible or reversible adhesion in heterogeneous environments. It is shown that, as in Taylor dispersion, the ratio of the channel half width b to the Brownian bacteria motility coefficient (D0 or dispersion coefficient) t(b)=b(2)/D(0) controls the different adhesion regimes along with the value of alpha. Universal scalings (with respect to dimensionless time t(*)=t/t(b)) for the mean position, =V(*)(eff)t(theta)(*), and mean-square displacement, =D(*)(eff)t(gamma)(*) exist for long-time dispersion and the coefficients were obtained. The model can account for a great many sorptive processes including reversible and irreversible sorption, and sub- and superdispersive regimes with just a few parameters.

Journal Article↗

[Dynamics of populations of microbial antagonists in nonsterile soil].

The interaction between populations was studied with Arthrobacter crystallopoietes and Streptomyces olivocinereus, an actinomycete producing the antibiotic heliomycin active against Gram-positive microorganisms. The two organisms were either cultivated together in a growth medium or the two populations were introduced simultaneously into nonsterile soil at different levels of population density. The antagonism was found in both cases: A. crystallopoietes cells died off when a population of the potential antagonist was added. The density of a population producing the antibiotic had to be sufficiently high for the antagonism to be manifested. The antagonism influenced the dynamics of a population of the antibiotic-sensitive microorganism. The results have confirmed earlier data to the effect that antibiotic synthesis is possible in nonsterile soil.

Anti-Bacterial Agents↗

Nisin Z, mutant nisin Z and lacticin 481 interactions with anionic lipids correlate with antimicrobial activity. A monolayer study.

Monomolecular layers of lipids at the air/water interface have been used as a model membrane to study membrane interactions of the lantibiotic nisin. The natural lantibiotics nisin A and nisin Z proved to have a high affinity for the anionic lipids phosphatidylglycerol and bis(phosphatidyl)glycerol (cardiolipin). The interaction with zwitterionic phopholipids or neutral lipids is very low at surface pressures higher than 32 mN/m. Nisin, nisin mutants and lacticin 481 show a remarkable correlation between anti-microbial activity and anionic lipid interaction. The results indicate that primarily the N-terminal part (residues 1-22) penetrates into the lipid phase. Reduction of the flexibility at positions 20-21 has a negative effect on monolayer interaction and activity. The C-terminal part is probably responsible for ionic interactions of nisin in monomeric or oligomeric form with anionic lipids. In mixtures of anionic and zwitterionic lipids maximal interactions are found at approximately 70 mol/100 mol anionic lipid. Gram-positive bacteria, which form the main target for nisin, are characterized by a high content of anionic lipids in the membrane. Monolayers formed of lipid extracts of bacteria sensitive to nisin were more strongly penetrated than those of bacteria relatively insensitive to nisin.

Amino Acid Sequence↗

Molecular genetics of bacterial attachment and biofouling.

Microbial adhesion to animate or inert surfaces is potentially mediated by nonspecific physical or specific ligand-receptor interactions. Growth and survival of the microbial community or biofilm then depends on adaptation to a series of changing environmental milieux. Within the realm of cell-cell interaction, recent advances suggest that flagella, fimbriae and other protein receptors are essential for bacterial attachment to surfaces. There has also been profound progress in the elucidation of genes and molecules necessary for bacterial attachments to surfaces and subsequent biofilm formation.

Bacterial Adhesion↗

Microbial community dynamics in nutrient-pulsed chemostats.

In nature, microbes are subject to nutrient fluxes. As the periodicity of nutrient flux lengthens, different physiological traits may be selected. The competitive exclusion principle stipulates that one organism will dominate these systems; however, interspecies interactions may produce a dynamic microbial community. These issues were investigated in chemostats pulsed with gelatin. Chemostats were run over 30 days with substrate addition continuously or at intervals of 0.5, 1 or 3 days. Growth rates were similar between pulse intervals. Ectoaminopeptidase activity levels remained relatively constant within a pulse interval. Bacterial community structure was monitored using denaturing gradient gel electrophoresis of PCR products of the 16S rRNA gene. There were dynamic changes at all periodicities; however, the pace of these changes decreased over time. Final communities were not identical between different treatments. The structure of persistent vs. active microbial populations was compared by denaturing gradient gel electrophoresis of the PCR and reverse transcriptase-PCR amplicons of 16S rDNA and rRNA templates, respectively. For all the chemostats, the rRNA profiles were not identical to the rDNA profiles for a sample. These experiments demonstrate that complex community dynamics can occur under environmental heterogeneities that are modest relative to those found in natural aquatic habitats. Furthermore, the physiological functionality of these dynamic communities was stable.

Aminopeptidases↗

The intestinal microflora: potentially fertile ground for microbial physiologists.

The intestinal microflora provides opportunities for microbial physiological research. The metabolic interactions of bacterial inhabitants of the intestinal community, bacterial bioenergetics, preferential utilization of substrates as energy sources by specific bacterial species, and intercellular signalling are among the topics of challenging research awaiting the attention of microbial physiologists.

Animals↗

Microbial Food Webs in Marine Sediments. II. Seasonal Changes in Trophic Interactions in a Sandy Tidal Flat Community

>AbstractThe role of grazing by marine sediment flagellates, ciliates, and meiobenthic animals in controlling production of their bacterial and diatom prey was investigated. At six selected time points, over the year, bacterial production and diatom standing stock were compared to grazing pressure exercised by proto- and micrometazoan consumers. The intensity of prey-predator relations showed pronounced yearly dynamics in which two stages could be distinguished. During the first phase, from the end of winter to mid-summer, the consumption of diatoms gradually increased, with possible overgrazing at the end of the period. This was followed by a collapse of diatom abundance, to the winter level. During the first stage, no appreciable bacterial consumption was observed in spite of the high abundance and production of bacteria. The second stage started in mid-summer and continued through the fall. During this period, the grazing on bacteria increased and reached the year's maximum. For at least a brief period (October), micrograzers removed the majority of bacterial production. In contrast, herbivory stayed at the year's lowest level, and diatoms appeared to be controlled by factors other than grazing. The observed ingestion rates seem to support the apparent energy requirements of flagellates and some ciliates (scuticociliates and hypotrichids). Other ciliates (pleurostomatids and karyorelictids) could not subsist on the observed diet and might have to complement it with other energy sources, possibly via dissolved organic matter absorption.

Journal Article↗