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[Community-acquired bacterial meningitis in the Loire-Atlantic region: evolution of pneumococcal and meningococcal sensitivity to penicillin].

OBJECTIVES: An epidemiological study of community-acquired bacterial meningitis was conducted in Loire-Atlantique in subjects aged over 1 month. Risk factors and changes in pneumococcal and meningococcal susceptibility to betalactams were analyzed. PATIENTS AND METHODS: All cases of proven or presumed bacterial meningitis registered by Loire-Atlantic bacteriology laboratories between May 1995 and April 1998 were analyzed. RESULTS: One hundred two cases were registered (annual incidence: 3.12 cases per 100,000 inhabitants). In children (33 cases) the main germs were meningococci (51%), pneumococci (24%) and Haemophilus influenzae (6%). In adults (69 cases), pneumococci (49%), meningococci (14%) and Listeria (4%) predominated. An underlying disease was noted 44% of the cases. Mortality was 17.6%. Sequellae were observed in 9.5%. Some degree of penicillin resistance was observed in 45% of the pneumococcal strains and in 50% of the meningococcal strains. Half of the pneumococcal strains were also resistant to third generation cephalosporins (C3G). No risk factor was significantly related to resistant strains. Susceptibility to antibiotics was not correlated with mortality for either pneumococcal or meningococcal strains, but sequellae were more frequent after meningitis caused by resistant pneumococci. CONCLUSION: For cases of community-acquired meningococcal meningitis diagnosed in 1999, it would be advisable to prescribe a combination C3G-vancomycin regimen as the first line empirical treatment while waiting for results of susceptibility tests. Certain guidelines proposed by the February 1996 consensus conference on community-acquired purulent meningitis would thus need to be amended.

Adult↗

Improvement of the lytic properties of a beta-1,3-glucanase by directed evolution.

BGLII is a bacterial endoglucanase that hydrolyzes the beta-1,3-glucan present in yeast cell walls, resulting in lysis of Saccharomyces cerevisiae. As a result of this property, BGLII is considered a potential tool for downstream processing and recovery of biotechnological products produced in yeast. Here we describe the improvement of the yeast lytic activity of BGLII, achieved by a directed evolution approach involving random mutagenesis and screening for variants with improved catalytic activity, combined with site-directed mutagenesis. A BGLII variant having three times the wild-type hydrolytic activity on laminarin was identified. The purified enzyme also exhibited higher lytic activity on yeast cells. Mutations causing the improvements are located very close to each other in the amino acid sequence, suggesting that the region should be considered as a target for further improvements of the glucanase activity. These results demonstrate the feasibility of molecular evolution methods for the improvement of the BGLII hydrolytic activity, and open a window for further improvement of this or other properties in glycosyl hydrolases in general.

Bacteria↗

[On bacterial origin of mitochondria in eukaryotes in the light of current ideas of evolution of the organic world].

The hypothesis of bacterial origin of mitochondria, which existed until the end of the 20th century, has been confirmed on the basis of the current concepts of organic world evolution in the open sea hydrosphere and original data on the entry of bacteria (prokaryotes0 in the cells of eukaryotes and their transformation into the mitochondrial mechanism of aerobic energy metabolism. This hypothesis can now be considered as a factually substantiated theory. The process of endocytosis of bacteria in the tissues of eukaryotes, which began at the onset of transition of the anaerobic state of open sea hydrosphere and land atmosphere (Early Proterozoic), is considered as the beginning of symbiotic mode of life of organisms of the Proterozoic and Postproterozoic organic world.

Aerobiosis↗

Congruent evolution between whiteflies (Homoptera: Aleyrodidae) and their bacterial endosymbionts based on respective 18S and 16S rDNAs.

Whiteflies (family Aleyrodidae) possess heritable eubacterial endosymbionts sustained in specialized organ-like structure called mycetomes. Comparisons of distances between the ash whitefly, Siphoninus phillyreae, and two biotypes ("A" and "B") of the sweetpotato whitefly, Bemisia tabaci, based on sequence analysis of genes for 18S rRNAs (rDNAs), were equivalent to the distances represented by the 16S rDNAs of their respective endosymbionts. This finding indicates that evolutionary divergence in whitefly hosts and their endosymbionts is congruent. The nucleotide sequences of the 18S rDNAs and endosymbiont 16S rDNAs indicate the two biotypes of B. tabaci are the same species.

Animals↗

Pathogenicity islands and virulence evolution in Listeria.

As in other bacterial pathogens, the virulence determinants of Listeria species are clustered in genomic islands scattered along the chromosome. This review summarizes current knowledge about the structure, distribution and role in pathogenesis of Listeria virulence loci. Hypotheses about the mode of acquisition and evolution of these loci in this group of Gram-positive bacteria are presented and discussed.

Amino Acid Sequence↗

Comparative genomic analysis of the sigB operon in Listeria monocytogenes and in other Gram-positive bacteria.

The stress-responsive, alternative sigma factor sigmaB has been described in members of three Gram-positive genera, Bacillus, Listeria, and Staphylococcus. In these bacteria, sigmaB appears to play an important role in facilitating rapid adaptation to and survival in stressful environments. sigmaB activity is regulated through a complex system of phosphatases and kinases encoded by rsb (regulator of sigma B) genes. We describe the sigB operon structure for the facultative intracellular pathogen Listeria monocytogenes and apply this sequence as well as other previously described sigB operon sequences to probe the evolution and functional conservation of the sigmaB stress response system among different Gram-positive bacteria. While sigmaB as well as two Rsbs (RsbS and RsbT) are highly conserved (73%, 84%, and 79% average amino acid [aa] identities, respectively), the predicted aa sequences of the other Rsb proteins showed less conservation (62-71% aa identities). Furthermore, the sigB operon structure varies among bacterial species. Bacterial species differ in the numbers and identities of rsb genes encoded in their genomes. We thus conclude that the sigmaB stress-response system as represented by the sigB operon has diverged in both its overall components as well as in the sequences of its individual proteins, even among closely related bacterial species. Differential evolution of this stress response system among various genera may represent a strategy that enables bacteria to adapt cellular response and survival systems to a variety of stress conditions.

Amino Acid Sequence↗

Monitoring of a pyrite-oxidising bacterial population using DNA single-strand conformation polymorphism and microscopic techniques.

The single-strand conformation polymorphism (SSCP) technique was used to study the evolution of a bacterial consortium during the batch oxidation of a cobaltiferous pyrite in two types of bio-reactor: a bubble column and a classical stirred tank. Sequencing 16S rDNA revealed the presence of three organisms affiliated to Leptospirillum ferrooxidans, Acidithiobacillus thiooxidans and Sulfobacillus thermosulfidooxidans, respectively. Attempts were made to determine the proportions of bacteria attached to solid particles or freely suspended in the medium using a combination of PCR-SSCP and a microscopic technique. Ac. thiooxidans-related bacteria were dominant in the liquid during the early phase of the batch, but were later supplanted by L. ferrooxidans-related bacteria. L. ferrooxidans-related organisms were always in the majority on the solids. The growth of S. thermosulfidooxidans-related bacteria seemed to be favoured by the bubble-column reactor.

Bacteria↗

Mutagenic topography of the E. coli chromosome.

The lactose fermenting genes in E. coli have been transposed to various chromosomal locations. The bacterial strains were mutagenized with different chemical mutagens and the frequency of Lac negative mutant colonies was measured as a function of lactose gene location in the chromosome. There appears to be a highly mutable location between 58-60 minutes on the E. coli map. This region does not appear to be correlated with the origin of DNA replication or with the terminus. The possible significance of this mutable region in the evolution of new bacterial genes is discussed.

Biological Evolution↗

Curiously modern DNA for a "250 million-year-old" bacterium.

Studies of ancient DNA have attracted considerable attention in scientific journals and the popular press. Several of the more extreme claims for ancient DNA have been questioned on biochemical grounds (i.e., DNA surviving longer than expected) and evolutionary grounds (i.e., nucleotide substitution patterns not matching theoretical expectations for ancient DNA). A recent letter to Nature from Vreeland et al. (2000), however, tops all others with respect to age and condition of the specimen. These researchers extracted and cultured a bacterium from an inclusion body from what they claim is a 250 million-year (Myr)-old salt crystal. If substantiated, this observation could fundamentally alter views about bacterial physiology, ecology and evolution. Here we report on molecular evolutionary analyses of the 16S rDNA from this specimen. We find that 2-9-3 differs from a modern halophile, Salibacillus marismortui, by just 3 unambiguous bp in 16S rDNA, versus the approximately 59 bp that would be expected if these bacteria evolved at the same rate as other bacteria. We show, using a Poisson distribution, that unless it can be shown that S. marismortui evolves 5 to 10 times more slowly than other bacteria for which 16S rDNA substitution rates have been established, Vreeland et al.'s claim would be rejected at the 0.05 level. Also, a molecular clock test and a relative rates test fail to substantiate Vreeland et al.'s claim that strain 2-9-3 is a 250-Myr-old bacterium. The report of Vreeland et al. thus falls into a long series of suspect ancient DNA studies.

Bacteria↗

Serum orosomucoid concentration in newborn infants.

Serum orosomucoid concentration was measured by laser nephelometry in 1970 serum samples collected from 1170 full term and preterm infants. The determinations were carried out in 1 h. Reference values are given: they show that the low levels at birth are influenced by gestational age. The concentrations increase rapidly during the first week in all infants, the adult values being reached by 10 months of age. High levels of orosomucoid concentration were detected in 85% of the infants with severe bacterial infections. Serum orosomucoid concentration proved less valuable in viral and parasitic infection. Twenty-six per cent of the sick infants without infection had a slightly elevated orosomucoid level which decreased rapidly. In the bacterial infections the evolution of serum orosomucoid concentration followed the clinical course. Thus serum orosomucoid concentration was a useful parameter for diagnosis and monitoring of bacterial infection in neonates.

Bacterial Infections↗

Mobile group II introns.

Mobile group II introns, found in bacterial and organellar genomes, are both catalytic RNAs and retrotransposable elements. They use an extraordinary mobility mechanism in which the excised intron RNA reverse splices directly into a DNA target site and is then reverse transcribed by the intron-encoded protein. After DNA insertion, the introns remove themselves by protein-assisted, autocatalytic RNA splicing, thereby minimizing host damage. Here we discuss the experimental basis for our current understanding of group II intron mobility mechanisms, beginning with genetic observations in yeast mitochondria, and culminating with a detailed understanding of molecular mechanisms shared by organellar and bacterial group II introns. We also discuss recently discovered links between group II intron mobility and DNA replication, new insights into group II intron evolution arising from bacterial genome sequencing, and the evolutionary relationship between group II introns and both eukaryotic spliceosomal introns and non-LTR-retrotransposons. Finally, we describe the development of mobile group II introns into gene-targeting vectors, "targetrons," which have programmable target specificity.

Bacteria↗

[Organization of the Escherichia coli genome].

A review of literature data reveals that for the last years, the molecular biology techniques have been of an increasing use in the study of the Escherichia coli genome, having supplemented the standard genetic mapping. For the proper understanding of the Escherichia coli genome organization, recombinational events occurring in the course of evolution should be considered. The bacterial genome seems to carry traces of both "long-term" evolution, possibly responsible for appearance of the bacterial cell itself, and "current" evolution, consisting mainly of periodic genome entering by new plasmid-originated genes. It is supposed that in the process of stabilization within a genome, every new gene undergoes a stage of the "transgene", that is the gene situated in a transposon on the chromosome. In parallel with integration of new genes into the genome, some genes deleting should also take place. The formation of deletions could occur by unequal crossing over in segments of direct homologous repeats which seem to be ordinarily revealed in the experimental study of the tandem gene duplications.

Chromosome Mapping↗

Ecotypes of the Mycobacterium tuberculosis complex.

A phylogeny of the Mycobacterium tuberculosis complex has recently shown that the animal-adapted strains are found in a single lineage marked by the deletion of chromosomal region 9 (RD9) [Brosch et al., 2002. A new evolutionary scenario for the Mycobacterium tuberculosis complex. Proc. Natl Acad. Sci. USA 99 (6), 3684-3689]. We have obtained the spoligotype patterns of the RD9 deleted strains used to generate this new evolutionary scenario and we show that the presence of spoligotype spacers 3, 9, 16, 39, and 40-43 is phylogenetically informative in this lineage. We have used the phylogenetically informative spoligotype spacers to screen a database of spoligotype patterns and have identified further members of a group of strains apparently host-adapted to antelopes. The presence of the spoligotype spacers is congruent with the phylogeny generated by chromosomal deletions, suggesting that recombination is rare or absent between strains of this lineage. The phylogenetically informative spacers, in concert with the previously identified single nucleotide mutations and chromosomal deletions, can be used to identify a series of clades in the RD9 deleted lineage each with a separate host preference. Finally, we discuss the application of the ecotype concept to this series of clades and suggest that the M. tuberculosis complex may best be described as a series of host-adapted ecotypes.

Animals↗

Genetics of the tryptophan biosynthetic pathway of the prokaryotic endosymbiont (Buchnera) of the aphid Schlechtendalia chinensis.

Two DNA fragments (3941 and 7152 base pairs) from the procaryotic endosymbiont (Buchnera) of the aphid Schlechtendalia chinensis were cloned and sequenced. The smaller fragment contained trpEG and the larger fragment contained trpDC(F)BA, genes coding for enzymes of the tryptophan biosynthetic pathway which convert chorismate to tryptophan. Both of these gene clusters were present as one copy on the endosymbiont chromosome and probably constitute two transcription units. The deduced amino acid sequences of the proteins was 51-61% identical to the corresponding proteins were previously studied in Buchnera of the aphid Schizaphis graminum. In this endosymbiont, trpEG is amplified and located on a plasmid, whereas, in the endosymbiont of S. chinensis, as in other eubacteria, trpEG occurs as a single copy on the bacterial chromosome. Implications of these findings for the evolution of this mutualistic association are discussed.

Amino Acid Sequence↗

Bacterial endosymbionts of insects: insights from comparative genomics.

The development of molecular techniques for the study of uncultured bacteria allowed the extensive study of the widespread association between insects and intracellular symbiotic bacteria. Most of the bacterial endosymbionts involved in such associations are gamma-proteobacteria, closely related to Escherichia coli. In recent years, five genomes from insect endosymbionts have been sequenced, allowing the performance of extensive genome comparative analysis that, as a complement of phylogenetic studies, and analysis on individual genes, can help to understand the different traits of this particular association, including how the symbiotic process is established, the explanation of the special features of these microbial genomes, the bases of this intimate association and the possible future that awaits the endosymbionts with extremely reduced genomes.

Adaptation, Biological↗

Evolution of transcription regulatory genes is linked to niche specialization in the bacterial pathogen Streptococcus pyogenes.

Streptococcus pyogenes is a highly prevalent bacterial pathogen, most often giving rise to superficial infections at the throat or skin of its human host. Three genotype-defined subpopulations of strains exhibiting strong tropisms for either the throat or skin (specialists) or having no obvious tissue site preference (generalists) are recognized. Since the microenvironments at the throat and skin are distinct, the signal transduction pathways leading to the control of gene expression may also differ for throat versus skin strains of S. pyogenes. Two loci (mga and rofA/nra) encoding global regulators of virulence gene expression are positioned 300 kb apart on the genome; each contains alleles forming two major sequence clusters of approximately 25 to 30% divergence that are under balancing selection. Strong linkage disequilibrium is observed between sequence clusters of the transcription regulatory loci and the subpopulations of throat and skin specialists, against a background of high recombination rates among housekeeping genes. A taxonomically distinct commensal species (Streptococcus dysgalactiae subspecies equisimilus) shares highly homologous rof alleles. The findings provide strong support for a mechanism underlying niche specialization that involves orthologous replacement of regulatory genes following interspecies horizontal transfer, although the directionality of gene exchange remains unknown.

Bacterial Proteins↗