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Identification of novel essential Escherichia coli genes conserved among pathogenic bacteria.

We deleted a subset of 27 open reading frames (ORFs) from Escherichia coli which encode previously uncharacterized, probably soluble gene products homologous to proteins from a broad spectrum of bacterial pathogens such as Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae and Enterococcus faecalis and only distantly related to eukaryotic proteins. Six novel bacteria-specific genes essential for growth in complex medium could be identified through a combination of bioinformatics-based and experimental approaches. We also compared our data to published results of gene inactivation projects with Mycoplasma genitalium and Bacillus subtilis and looked for homologs in all known prokaryotic genomes. Such analyses highlight the enormous metabolic flexibility of prokaryotes. Six of 27 studied genes have been functionally characterized up to now, amongst these four of the essential genes. The gene products YgbP, YgbB and YchB are involved in the non-mevalonate pathway of isoprenoid biosynthesis. KdtB is characterized as the posphopantetheine adenylyltransferase CoaD. There are indications that the other two essential gene products YjeE and YqgF, which we have identified, also possess enzymatic functions. These findings demonstrate the potential of such proteins to be used in screening of large chemical libraries for inhibitors which could be further developed to novel broad-spectrum antibiotics.

Bacillus subtilis↗

[Identification of the mRNA of the nucleocapsid proteins of pathogenic arenaviruses].

Total RNA from cells infected with Machupo and Lassa viruses as well as individual sedimentation classes of these RNAs were translated in the cell-free protein-synthesizing system from rabbit reticulocytes. The translation products were precipitated either with anti-Machupo immune gamma-globulin or monoclonal antibodies to nucleocapsid protein (NP) of Lassa virus. Both total RNA and RNA fraction with the sedimentation coefficient 15-16S promoted the synthesis of protein which comigrated in gel with NP protein of purified virions. It is concluded that monocistron mRNA for NP protein of arenaviruses has the sedimentation coefficient 15-16 S.

Animals↗

The traveller and emerging infections: sentinel, courier, transmitter.

The movement of populations shapes the patterns and distribution of infectious diseases globally. The consequences of travel are seen in the traveller and in places and populations visited and may persist long after travel. The traveller can be seen as an interactive biological unit who picks up, processes, carries and drops off microbial genetic material. A traveller can introduce potential pathogens in the absence of signs or symptoms of illness. Travellers can serve as a sentinel population; study of them can provide insights into the presence and level of risk of transmission of infections in other geographical regions. Travellers can also be seen as couriers who inadvertently ferry pathogens and microbial genetic material to regions where researchers can carry out detailed analyses that can help to map the location and movement of strains, genotypes and resistance patterns. The laboratory plays a key role in the identification and characterization of pathogens, which can inform management of individual patients and the public health response. The connectedness and mobility in the world today facilitate the emergence of infectious diseases in humans and also in animals and plants. Many traditional barriers have been breached by travel, roads and technology. Population size and density favour spread of many infections. The rapid generation time of microbes and their capacity to adapt to changes in the physico-chemical and immunological environment will pose continuing challenges.

Animals↗

[Primary investigation on variable but nonculturable-state of enterotoxigenic Escherichia coli in vitro].

OBJECTIVE: 7 variable but nonculturable-state strains of Enterotoxigenic Escherichia coli (ETEC) during the routine bacterial subculture were found in our lab and their morphology and antigen studied. Biological features, antigens and pathogenicity of the revertants were also tested and compared to that of the initial strains in order to detect their variations. METHODS: Biological variations between the variable but nonculturable-state and the revertant of every strain were detected, using the routine gram-staining, reverting the isolates in animal intestinal, reverting their pathogenicity, serological agglutination, biochemical identifications and antibiotic resistance tests. RESULTS: For the 7 variable but nonculturable-state strains of ETEC,other than the trains that had changed into sphero vegetale cells, there were no other obvious variations found. However, high pathogenicity of these strains still remained. CONCLUSION: The presence of variable but nonculturable-state strains suggested that the routine method of bacteria storage should be changed and more attention should be paid to realize the existence of this kind of bacteria during the routine surveillance of the communicable diseases.

Antigens, Bacterial↗

Construction of replicative and integrative plasmids for setting up the in vivo expression technology in Helicobacter pylori.

Some pathogenic factors of Helicobacter pylori, a bacterium involved in peptic ulcer and gastric cancer, have already been identified using either global or particular approach, but there are still some orphan genes and unidentified pathogenic factors. One of the methods used successfully for the identification of virulence genes of many pathogens is the in vivo expression technology. We describe here the construction and sequences of three different plasmids, one integrative and two replicatives, for the identification of virulence genes by using in vivo expression technology in H. pylori, and of potential use in other bacteria such as Campylobacter spp. Moreover, the use of the green fluorescent protein could allow to classify the genes according to the strength of their expression and to identify those which are repressed upon interaction with gastric mucosa.

DNA, Bacterial↗

[DNA microarrays and their application in detecting and identifying intestinal pathogens].

DNA microarrays offer many advantages of high throughout, automation, rapid detection, and so on. Therefore, this technology had been used in many fields such as molecular epidemiology of bacteria, microbial gene identification, disease mechanism, gene mutation, gene expression identification, DNA sequencing and medicine screening etc. The assays for identifying pathogens using DNA microarrays reported aboard recently are introduced. The application of DNA microarrays in detecting and identifying intestinal pathogens mainly includes three aspects: the identification of toxin and characteristic genes of pathogens, the identification of bacterial DNA or RNA directly, the simultaneous detection of a large number of intestinal pathogens with the target - gene of ribosomal RNA. Because of its high efficiency, DNA microarrays is superior to other biological method. Obviously DNA microarrays technology may be useful in identifying intestinal pathogens and have a wide prospect.

Animals↗

[Evaluation of the relation between the traditional index on the identification of Bacillus anthracis and its virulence determinant genes].

OBJECTIVE: To evaluate the significance of traditional index on the identification of Bacillus anthracis and its correlation with pathogenic strains. METHODS: Routine bacteriologic methods and PCR of virulence genes were used to determine the difference on traditional identification index between pathogenic and nonpathogenic Bacillus. RESULTS: The characteristics of colony, with hemolysis and mobility both negative are the important interrelated characters of pathogenic strains of Bacillus anthracis. CONCLUSION: To judge the risk of anthrax, virulence of genes must be first defined. Some traditional methods for identification are still useful when molecular biological methods are not available.

Animals↗

Evaluation of the IDS RapID SS/u system for rapid identification of urinary microorganisms.

The accuracy of a new rapid identification system for common urinary pathogens was compared with that of conventional methods and of miniaturized 18-24-hour identification panels. The rapid system, RapID SS/u (Innovative Diagnostic System Inc., Atlanta, GA) is a non-growth-dependent micro-method that identifies selected gram-negative bacilli, gram-positive cocci, and yeasts in two hours by detection of constitutive enzymes acting on chromogenic substrates. A total of 185 representative clinical urinary isolates were tested, including 24 gram-positive cocci, 140 gram-negative bacilli, and 21 yeasts. Identifications by the rapid system were compared with the ones obtained by reference conventional methods for gram-positive cocci and yeasts. For gram-negative bacilli, identifications were compared with the ones obtained by MicroScan Combo Panel (American MicroScan, Mahwah, NJ), and all discrepancies were resolved by testing with API 20E (Analytab Products, Plainview, NY). Overall, the RapID SS/u system correctly identified to genus 160 of 185 isolates (86.5%). For 14 additional isolates (7.6%) the system provided probability overlap identifications that required further testing. Two (1%) isolates failed to be identified, and nine isolates (4.9%) were misidentified by the system. Discrepancies involved five strains of Citrobacter, one Enterobacter, one Morganella, and one Providencia species. The authors conclude that the RapID SS/u system provided rapid and accurate genus identification of most microorganisms commonly isolated from urine.

Humans↗

Presence of potentially pathogenic Babesia sp. for human in Ixodes ricinus in Switzerland.

We have designed and performed a new PCR method based on the 18S rRNA in order to individuate the presence and the identity of Babesia parasites. Out of 1159 Ixodes ricinus (Acari: Ixodidae) ticks collected in four areas of Switzerland, nine were found to contain Babesia DNA. Sequencing of the short amplicon obtained (411-452 bp) allowed the identification of three human pathogenic species: Babesia microti, B. divergens, for the first time in Switzerland, Babesia sp. EU1. We also report coinfections with B. sp. EU1-Borrelia burgdorferi sensu stricto and Babesia sp. EU1-B. afzelii.

Animals↗

The impact of host genetics on susceptibility to human infectious diseases.

The development of genetic epidemiology methods using recent human genetic mapping information, together with the growing availability of candidate genes, has led to major advances in the identification of host genes involved in human infectious diseases. Within the past year, highlights include the mapping of a locus controlling the intensity of infection by Schistosoma mansoni, the demonstration that mutations in the interferon-gamma receptor 1 gene are causative of disseminated infection due to weakly pathogenic mycobacteria, and the identification, in the CCR5 gene, of a deletion which provides high protection against HIV-1 infection. The impact of these findings on the understanding of infectious disease pathogenesis and on the design of future preventive and therapeutic strategies should be considerable.

Animals↗

Identification and ribotypes of Staphylococcus caprae isolates isolated as human pathogens and from goat milk.

We report five cases of human infection with Staphylococcus caprae. Two were community acquired (one case each of endocarditis and urinary tract infection); the other three were acquired in a hospital (two cases of bacteremia associated with intravenous access and one case of urinary tract infection). Analysis of human isolates and goat isolates from eight herds showed that they could be misidentified by some commercial identification systems but were clearly identified as S. caprae by ribotyping, according to their species-specific ribotype. Phylogenetic methods applied to the ribotypes did not reveal two distinct lineages corresponding to the goat and human origins of the isolates, although human ribotypes were clearly distinguishable by the presence of a core of four specific bands. The latter observation may reflect some degree of evolutionary change within the species between human and goat isolates.

Adult↗