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Bacillus anthracis multiplication, persistence, and genetic exchange in the rhizosphere of grass plants.

Bacillus anthracis, the causative agent of anthrax, is known for its rapid proliferation and dissemination in mammalian hosts. In contrast, little information exists regarding the lifestyle of this important pathogen outside of the host. Considering that Bacillus species, including close relatives of B. anthracis, are saprophytic soil organisms, we investigated the capacity of B. anthracis spores to germinate in the rhizosphere and to establish populations of vegetative cells that could support horizontal gene transfer in the soil. Using a simple grass plant-soil model system, we show that B. anthracis strains germinate on and around roots, growing in characteristic long filaments. From 2 to 4 days postinoculation, approximately one-half of the B. anthracis CFU recovered from soil containing grass seedlings arose from heat-sensitive organisms, while B. anthracis CFU retrieved from soil without plants consisted of primarily heat-resistant spores. Co-inoculation of the plant-soil system with spores of a fertile B. anthracis strain carrying the tetracycline resistance plasmid pBC16 and a selectable B. anthracis recipient strain resulted in transfer of pBC16 from the donor to the recipient as early as 3 days postinoculation. Our findings demonstrate that B. anthracis can survive as a saprophyte outside of the host. The data suggest that horizontal gene transfer in the rhizosphere of grass plants may play a role in the evolution of the Bacillus cereus group species.

Bacillus anthracis↗

First characterization of a cluster of VanA-type glycopeptide-resistant Enterococcus faecium, Colombia.

From August 1998 to October 1999, glycopeptide-resistant enterococci (GRE) were isolated from 23 infected patients at a teaching hospital in Medellín, Colombia. Identification at the species level and by multiplex polymerase chain reaction assay indicated that all isolates were Enterococcus faecium. The isolates were highly resistant to ampicillin, ciprofloxacin, gentamicin, penicillin, streptomycin, teicoplanin, and vancomycin; they were susceptible only to chloramphenicol, linezolid, and nitrofurantoin. Determination of glycopeptide genotype indicated the presence of the vanA gene in all isolates. Molecular typing by pulsed field gel electrophoresis showed that all isolates were closely related. This study is the first molecular characterization of GRE in Colombia.

Anti-Bacterial Agents↗

CD1 proteins: targets of T cell recognition in innate and adaptive immunity.

The CD1 family consists of antigen presenting molecules encoded by genes located outside of the major histocompatibility complex. CD1 proteins are conserved among mammalian species and are expressed on the surface of cells involved in antigen presentation. The CD1 system has been shown to be involved in activation of cell-mediated responses, and T cells specific for either CD1 molecules or antigens presented by CD1 have been isolated. Structural and biochemical analyses demonstrate that antigens presented by CD1 are nonpeptide lipid or glycolipid structures, including examples found in the cell walls of pathogenic mycobacteria. The hydrophobic part of these antigens most likely binds in the CD1 ligand-binding groove, whereas the polar headgroup of these antigens appears to make direct contact with the T cell receptor and determines specific recognition. Presentation of antigens by CD1 molecules requires uptake and intracellular processing by antigen presenting cells and can be achieved for both exogenous and endogenous antigens. T cells recognizing CD1 restricted antigens have a broad range of functional activities that suggest that the CD1 system is involved in both innate and adaptive immune responses against microbial infections.

Animals↗

[Increased incidence of some strains of Klebsiella aerogenes].

Study of the incidence of Klebsiella aerogenes strains in neonates and infants with acute enterocolitis in the Maternity and Pediatric Departments gave the following results: - bacteriologic investigations were carried out in 1831 subjects, isolating 138 Klebsiella aerogenes strains; - the proportion of Klebsiella isolations increased from 4.6% in 1973 to 11.1% in 1974; - the maximum receptivity to Klebsiella aerogenes was noted in the 0-2 years age group, in a proportion of 23%; - in the 0-6 months age group Klebsiella aerogenes was the evident causal agent of enterocolitis, being isolated in pure cultures or as predominant germ in 70-80% of the cases. Worthy of note was the increased resistance to antibiotics in terms of the rising age-group scale. The predominance of a single serotype (K 15), the similar sensitivity of the strains to phage 32 and to antibiotics plead for an epidemic evolution of the cases studied.

Adult↗

Processing dewatered sewage sludge using electrokinetic technology.

High content of heavy metals and presence of pathogens in the dewatered sewage sludge have been the main obstacles for land application of sewage sludge-made fertilizer. The aim of this study was to examine the effects of the innovative electrokinetic (EK) technology on removal of heavy metals from sewage sludge, on the reduction of pathogens, and on sludge chemical characteristics. The results showed that the removal efficiencies for Zn, Cu, Ni, Cr, As and Pb were 94.9%, 95.4%, 89.7%, 67.8%, 31.2% and 18.7%, respectively. Acidification pretreatment of the dewatered sludge for 29 h decreased the content of heterotrophic bacteria from 1.5 x 10(8) c.f.u./g of wet sludge to 1.1 x 10(4) c.f.u./g of wet sludge. Although the initial content of total coliforms and fecal coliforms in sewage sludge were 5.8 x 10(5) c.f.u./g of wet sludge and 4.0 x 10(5) c.f.u./g of wet sludge, respectively, no viable cells were detected. Minor losses of K and N were detected, but the loss of P was found to be significant in EK treated sewage sludge. The treated sludge was technically considered as very stable based on the carbon dioxide evolution rate.

Bioreactors↗

Phylogeny, function, and evolution of the cupins, a structurally conserved, functionally diverse superfamily of proteins.

The cupin superfamily is a group of functionally diverse proteins that are found in all three kingdoms of life, Archaea, Eubacteria, and Eukaryota. These proteins have a characteristic signature domain comprising two histidine- containing motifs separated by an intermotif region of variable length. This domain consists of six beta strands within a conserved beta barrel structure. Most cupins, such as microbial phosphomannose isomerases (PMIs), AraC- type transcriptional regulators, and cereal oxalate oxidases (OXOs), contain only a single domain, whereas others, such as seed storage proteins and oxalate decarboxylases (OXDCs), are bi-cupins with two pairs of motifs. Although some cupins have known functions and have been characterized at the biochemical level, the majority are known only from gene cloning or sequencing projects. In this study, phylogenetic analyses were conducted on the conserved domain to investigate the evolution and structure/function relationships of cupins, with an emphasis on single- domain plant germin-like proteins (GLPs). An unrooted phylogeny of cupins from a wide spectrum of evolutionary lineages identified three main clusters, microbial PMIs, OXDCs, and plant GLPs. The sister group to the plant GLPs in the global analysis was then used to root a phylogeny of all available plant GLPs. The resulting phylogeny contained three main clades, classifying the GLPs into distinct subfamilies. It is suggested that these subfamilies correlate with functional categories, one of which contains the bifunctional barley germin that has both OXO and superoxide dismutase (SOD) activity. It is proposed that GLPs function primarily as SODs, enzymes that protect plants from the effects of oxidative stress. Closer inspection of the DNA sequence encoding the intermotif region in plant GLPs showed global conservation of thymine in the second codon position, a character associated with hydrophobic residues. Since many of these proteins are multimeric and enzymatically inactive in their monomeric state, this conservation of hydrophobicity is thought to be associated with the need to maintain the various monomer- monomer interactions. The type of structure-based predictive analysis presented in this paper is an important approach for understanding gene function and evolution in an era when genomes from a wide range of organisms are being sequenced at a rapid rate.

Amino Acid Sequence↗

[Microbial sensitivity of Escherichia coli in community-acquired urinary tract infections].

OBJECTIVE: For effective empiric therapy of urinary tract infections in the extra-hospital setting the susceptibility pattern of uropathogens should be considered. Moreover, the evolution in sensitivity can be observed when comparing with susceptibility patterns in the previous years. This paper presents an analysis of our experience with Escherichia coli. MATERIAL AND METHODS: During 2002 and 1998, 895 and 595 strains of Escherichia coli respectively, isolated from extrahospitalary bacteriurias were collected in ten health centers in Bierzo (León, Spain). Sensitivity to nine most commonly antibiotics used in the clinical practise was determined. The existence of significant differences of susceptibility among years (2002-1998) was analyzed by the chi square test. RESULTS: Escherichia coli accounted for 63.4% of all isolates in 2002 and 50.8% in 1998. The prevalence of in-vitro susceptibilities to antibiotics were (2002-1998): fosfomycin (99.2%-99.3%; p = NS*), cefixime (98.3%-92.9%; p < 0.001), cefuroxime (96.5%-94.1%; p < 0.05), nitrofurantoin (94.5%-86.9%; p < 0.001), amoxycillin-clavulanic acid (93.1%-90.1%; p < 0.05), ciprofloxacin (77.1%-81.6%; p < 0.05), norfloxacin (75.8%-80.3%; p < 0.05), cotrimoxazole (71.5%-73.4%; p = NS*) and ampicillin (44%-41.4%; p = NS*). (*NS = No significant differences). CONCLUSION: The knowledge of the sensitivity of uropathogens to antimicrobians in a specific medium can allow us to use antibiotics rationally and initiate empirical therapy.

Anti-Bacterial Agents↗

Temporal and dose-dependent relationships between in vivo B cell receptor-targeted proliferation and deletion-induced by a microbial B cell toxin.

The effective functioning of the adaptive immune system requires careful clonal regulation within the B cell compartment. Some microbial pathogens produce virulence factors, like staphylococcal protein A, which interact at high frequencies with B lymphocyte through unconventional binding sites in BCR variable region frameworks conserved during evolution. We have characterized the in vivo effect of staphylococcal protein A treatment on peripheral B cells bearing susceptible BCR, and found a dose-dependent direct relationship over the range of 2 mg to <0.2 microg in the magnitude of induced BCR-targeted supraclonal cell death. Significantly, some level of targeted B cell proliferation was always detectable, with greatest interim supraclonal expansion demonstrated at 2 days after 20-microg treatment. Subsequently, this transient expansion always collapsed. In direct comparisons, i.p. treatment was more efficacious than i.v. treatment, although at higher doses this finding was less marked. These studies elucidate a general paradigm in which in vivo encounters with a B cell superantigen are uniformly associated with proliferative expansion followed by deletion that is more rapid and complete with higher doses, whereas lower doses lead to greater transient in vivo expansion with delayed deletion to levels at later times that are still quantitatively proportional to the dose. Our results document the potent in vivo B cell-targeted properties of a microbial B cell superantigen, even at submicrogram doses associated with great molar excess of circulating Ig, and clearly illustrate the intertwined relationships between targeted proliferative cycling and apoptotic death that is induced by a microbial B cell superantigen.

Animals↗

Cellular events in alveolitis and the evolution of pulmonary fibrosis.

"Alveolitis", as opposed to "pneumonia" sensu strictiori, is a term used to denote diffuse inflammatory changes of the pulmonary parenchyma, excluding those that result from local bacterial, fungal or other extracellular microbial growth. The various types of alveolitis are classified according to their histological characteristics and range from "luminal phagocytic" or "mural lymphoplasmacellular" and "exudative" to "fibrosing" alveolitis. In this overview, various exogenous and endogenous causes of different types of alveolitis, and the cellular events in their pathogenesis are briefly discussed to illustrate the complex mechanisms involved. Particular emphasis is placed on the possible transition from diffuse exudative to fibrosing alveolitis. It appears that pulmonary fibrosis, which is usually patchy rather than truly diffuse, does not have a uniform pathogenesis. Besides the possibility of a certain degree of a diffuse fibrosis three major pathways are evident: (1) granulation tissue budding into alveolar lumina (luminal fibrosis) (2) exudate incorporation into alveolar walls (mural fibrosis) and--at least equally important--(3) so-called collapse (atelectatic) induration (obliterative-interseptal fibrosis), a process that has largely been neglected so far.

Air Pollutants↗

Phytoalexins and disease resistance mechanisms from a perspective of evolution and adaptation.

Plants respond to cellular injury and infection by accumulating low molecular weight antimicrobial stress metabolites called phytoalexins. The accumulation of phytoalexins, together with lignification, suberization, callose formation and the production of agglutinins and inhibitors of extracellular microbial hydrolases, appears to be part of a multi-component response mechanism associated with disease resistance and wound repair. Compared to the antibody-antigen response in animals, the phytoalexin response in plants has low specificity for induction and activity of the phytoalexins. Plants also contain preformed antimicrobial chemical and physical barriers to infection in their external tissues. The successful pathogen has evolved to cope with preformed inhibitors and barriers and either avoids eliciting the response mechanism, or suppresses the mechanism, or detoxifies its antimicrobial components. Annual plants can be systemically immunized against diseases caused by viruses, bacteria and fungi by limited infection with any one of the respective organisms. As with animals, disease resistance in plants depends on the rate and magnitude of response rather than on the ability to respond. The genetic information for disease resistance is found in all organisms, and disease resistance is the rule in nature. The interactions of plants with microorganisms in their environment are nature's example of diplomacy--compromise, adjustment to change and avoidance of deadly conflict.

Adaptation, Biological↗

Modeling of yeast metabolism and process dynamics in batch fermentation.

Much is known about yeast metabolism and the kinetics of industrial batch fermentation processes. In this study, however, we provide the first tool to evaluate the dynamic interaction that exists between them. A stoichiometric model, using wine fermentation as a case study, was constructed to simulate batch cultures of Saccharomyces cerevisiae. Five differential equations describe the evolution of the main metabolites and biomass in the fermentation tank, while a set of underdetermined linear algebraic equations models the pseudo-steady-state microbial metabolism. Specific links between process variables and the reaction rates of metabolic pathways represent microorganism adaptation to environmental changes in the culture. Adaptation requirements to changes in the environment, optimal growth, and homeostasis were set as the physiological objectives. A linear programming routine was used to define optimal metabolic mass flux distribution at each instant throughout the process. The kinetics of the process arise from the dynamic interaction between the environment and metabolic flux distribution. The model assessed the effect of nitrogen starvation and ethanol toxicity in wine fermentation and it was able to simulate fermentation profiles qualitatively, while experimental fermentation yields were reproduced successfully as well.

Adaptation, Physiological↗

Modeling bacterial species abundance from small community surveys.

Taxon abundance patterns are a central focus in evolution and ecology, providing the basic architecture of natural assemblages and potential clues to their formative processes. To better interpret species abundance patterns in natural microbial communities, we examined the consequences of three fundamental types of abundance patterns--uniform, geometric, and lognormal distributions. Theoretical communities were constructed based on the three pattern types with 2000 to 20,000 species and 10(7) to 10(10) individuals. The "dominant" species (species 1) among models that differed only in pattern type generally varied in abundance by 1 to 3 orders of magnitude. At the extremes among all the models examined, the dominant species comprised as much as 16% and as little as 0.005% of the total community. Analysis of the models and comparison with seven published surveys suggests that entire soil bacterial communities do not routinely exhibit Preston's cannonical subset of lognormal distributions. Use of the models to evaluate survey limitations showed that common diversity indices are generally sensitive to sample size over the range (50 to 200 clones) commonly used for microbial communities, emphasizing the need to compare surveys of similar size. The results collectively demonstrate that uniform, geometric, and lognormal distributions have profoundly different experimental and ecological consequences. Further, defined abundance models provide a simple quantitative tool for evaluating abundance patterns in clone libraries (even small ones) from natural communities.

Bacteria↗

Evaluation of some biological tests as parameters for microbial activities in soils. I. Laboratory experiments.

The experiments were confined to the effects of the addition of different sources of carbon (glucose, wheat straw, and sawdust) on the microbial activities in soils: loamy sand, loam and saline clay were used. The parameters used for this purpose were total bacterial counts, dehydrogenase test, CO2 evolution, and oxidation of organic carbon. Salinity of soils had deleterious effects on all the parameters used. Addition of organic matter resulted in marked increases in bacterial counts, formazan production, and CO2 evolution. Glucose showed the most rapid effect and highest levels. Wheat straw yielded the highest results and sawdust always showed the least effect.

Bacteria↗