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Biomedical subjects

Didier Raoult

Publications and source records attributed to Didier Raoult.

At least 37 records · Page 2Linked to original sources

Quantitative histological examination of bioprosthetic heart valves.

BACKGROUND: The histological features that characterize infective endocarditis in bioprosthetic valves are not accurately defined. Moreover, bioprosthetic valves may have a noninfective, degenerative evolution associated with calcifications, vegetation-like lesions, and inflammatory infiltrates. Such histological findings may be misdiagnosed as infective endocarditis. METHODS: Pathologic analysis of inflamed bioprosthetic valve tissues was conducted retrospectively for 21 patients who underwent surgical removal of a bioprosthetic valve because of suspected infective endocarditis and for 67 patients who underwent surgical removal of a bioprosthetic valve because of noninfective dysfunction. To better define the histological criteria for infective endocarditis, we used quantitative image analysis to compare these 2 groups of patients with respect to vegetations, calcifications, and patterns of inflammation. RESULTS: Histologically, infective endocarditis in patients with bioprostheses was characterized by demonstration of microorganisms, vegetations, and neutrophil-rich, inflammatory infiltrates. Valve tissue specimens from patients whose bioprosthetic valves were removed because of noninfective complications showed, in 30% of cases, inflammatory infiltrates mainly composed of macrophages and lymphocytes. Inflammatory adherent thrombi that can occur to the surface of noninfective degenerative bioprostheses are differentiated because their vegetations have macrophage-rich content. A neutrophil surface area with a cutoff value of > or =1.5% of the total valve tissue surface area is highly specific (94%) for infective endocarditis. CONCLUSIONS: When no microorganisms are detected and vegetations are not found in bioprosthetic valve tissues during the histological examination, a neutrophil-rich inflammation might better define the term "active endocarditis" in the Duke criteria and would allow differentiation between infective endocarditis and inflammatory, noninfective valve processes in patients with bioprosthetic valves.

Adult↗

Comparative genomics of multidrug resistance in Acinetobacter baumannii.

Acinetobacter baumannii is a species of nonfermentative gram-negative bacteria commonly found in water and soil. This organism was susceptible to most antibiotics in the 1970s. It has now become a major cause of hospital-acquired infections worldwide due to its remarkable propensity to rapidly acquire resistance determinants to a wide range of antibacterial agents. Here we use a comparative genomic approach to identify the complete repertoire of resistance genes exhibited by the multidrug-resistant A. baumannii strain AYE, which is epidemic in France, as well as to investigate the mechanisms of their acquisition by comparison with the fully susceptible A. baumannii strain SDF, which is associated with human body lice. The assembly of the whole shotgun genome sequences of the strains AYE and SDF gave an estimated size of 3.9 and 3.2 Mb, respectively. A. baumannii strain AYE exhibits an 86-kb genomic region termed a resistance island--the largest identified to date--in which 45 resistance genes are clustered. At the homologous location, the SDF strain exhibits a 20 kb-genomic island flanked by transposases but devoid of resistance markers. Such a switching genomic structure might be a hotspot that could explain the rapid acquisition of resistance markers under antimicrobial pressure. Sequence similarity and phylogenetic analyses confirm that most of the resistance genes found in the A. baumannii strain AYE have been recently acquired from bacteria of the genera Pseudomonas, Salmonella, or Escherichia. This study also resulted in the discovery of 19 new putative resistance genes. Whole-genome sequencing appears to be a fast and efficient approach to the exhaustive identification of resistance genes in epidemic infectious agents of clinical significance.

Acinetobacter Infections↗

Identification of candidate antigen in Whipple's disease using a serological proteomic approach.

Whipple's disease (WD) is a chronic multisystemic infection, caused by Tropheryma whipplei, a Gram-positive rod. Recently, a reliable method has been developed for cultivating T. whipplei in vitro. This together with the availability of complete genome sequence of T. whipplei prompted us to initiate proteome analysis of T. whipplei. The objective of the present study was to identify candidate proteins for serological diagnosis of WD. Immunoreactivities of sera collected from 18 patients with WD were compared with those of 24 control subjects who did not have WD. For this, we used 2-DE, immunoblotting, and MS. In total, we identified 23 candidate antigenic proteins. These included a subset of six proteins, each of which was found significantly more frequently in cases as compared to their controls. The remaining 17 proteins were found exclusively in cases. The methods we used in the current study enabled us to identify candidate antigens that, in our view, might be useful for serological diagnosis of WD.

Actinomycetales↗

Yersinia pestis as a telluric, human ectoparasite-borne organism.

The classic epidemiological model of plague is an infection of rodents that is transmitted to human beings by rodent ectoparasites. This model fits with observations of sporadic and limited outbreaks, but hardly explains the persistence of plague foci for millennia or the epidemiological features drawn from the descriptions of historical pandemics. A comprehensive review of the published data, including scientific papers published in France between 1920 and 1940, allows the completion of the epidemiological chain by introducing soil as a reservoir, burrowing rodents as a first link, and human ectoparasites as the main driving force for pandemics. Modern studies are needed to confirm the validity of this controversial model and to assess the relative contribution of each link in the various epidemiological presentations of plague. If confirmed, these data should be taken into account to update public-health policies and bioterrorism risk management, particularly among ectoparasite-infested people.

Animals↗

The two faces of interleukin 10 in human infectious diseases.

Resolution of infections depends on the host's ability to mount a protective immune response. However, an exacerbated response to infections may result in deleterious lesions. Consequently, immunoregulatory mechanisms are needed to control immune response and prevent infection-associated lesions. Interleukin 10 may be a major regulator of innate and adaptive immunity in vitro and in animals, but its role in human infections is still unclear. Review of the published work reveals wide involvement of interleukin 10 in two major features of infectious diseases. On one hand, interleukin 10 prevents the development of immunopathological lesions that result from exacerbated protective immune response to acute and chronic infections. On the other hand, it is critically involved in persistence of bacteria and viruses by interfering with innate and adaptive protective immunity. Moreover, infections induce the expansion of interleukin-10-producing regulatory cells that are involved in protection against allergic diseases.

Bacterial Infections↗

Sub-acute neuropathy in patients with African tick bite fever.

African tick bite fever (ATBF) caused by Rickettsia africae is an emerging health problem in travellers to sub-Saharan Africa. We here present 6 patients with evidence of long-lasting sub-acute neuropathy following ATBF contracted during safari trips to southern Africa. Three patients developed radiating pain, paresthaesia and/or motor weakness of extremities, 2 had hemi-facial pain and paresthaesia, and 1 developed unilateral sensorineural hearing loss. When evaluated 3-26 months after symptom onset, cerebrospinal fluid samples from 5 patients were negative for R. africae PCR and serology, but revealed elevated protein content in 3 and mild pleocytosis in 1 case. Despite extensive investigations, no plausible alternative causes of neuropathy could be identified. Treatment with doxycycline in 2 patients had no clinical effect. Given the current increase of international safari tourism to sub-Saharan Africa, more cases of sub-acute neuropathy following ATBF may well be encountered in Europe and elsewhere in the y to come.

Adult↗

Are clinical isolates of Pseudomonas aeruginosa more virulent than hospital environmental isolates in amebal co-culture test?

OBJECTIVE: Pseudomonas aeruginosa is an important opportunistic pathogen in hospital-acquired infections. As an environmental bacterium, it colonizes soil and water as well as hospital water supply and shares its natural habitat with free-living amebae. It has been shown previously that an ameba host system can be used to analyze the virulence of P. aeruginosa strains. In this study we have developed a simple amebal co-culture test with Acanthamoeba polyphaga to compare the virulence of P. aeruginosa between human and environmental isolates. DESIGN: Experimental study. SETTING: The four state hospitals Intensive of Marseille. INTERVENTIONS: Eighty-one environmental strains isolated from the hospital water and 69 clinical isolates of P. aeruginosa (52 from blood cultures and 17 from bronchoalveolar lavage) were tested in the amebal co-culture test. MEASUREMENTS AND MAIN RESULTS: The results show that compared with environmental isolates, the isolates of P. aeruginosa from blood cultures were significantly more virulent in the ameba assay (37% vs. 62%, p=.006). CONCLUSIONS: These data suggest that clinical infections due to Pseudomonas aeruginosa are due, at least in part, to bacterial virulence. Furthermore, even within the constraints of small sample size, virulence of respiratory isolates is significantly correlated with the occurrence of P. aeruginosa bacteremia. Our results, as those published previously, suggest that at the strain level, acquisition of antimicrobial resistance is correlated with the diminution of virulence. However, at the population level, clinical isolates are more virulent as well as more frequently resistant to antibiotics, presumably due to acquisition of strains mostly through cross-contaminations.

Acanthamoeba↗

Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes.

Determination of the DNA G+C content of prokaryotic genomes using traditional methods is time-consuming and results may vary from laboratory to laboratory, depending on the technique used. We explored the possibility of extrapolating the genomic DNA G+C content of prokaryotes from gene sequences. For this, 127 universally conserved genes were studied from 50 prokaryotic genomes in the Clusters of Orthologous Groups database. Of these, 57 genes were present as a single copy in the genomes of 157 different prokaryote species available in GenBank. There was a strong correlation [coefficient of determination (r2) >95 %] between the DNA G+C contents of 20 genes and their corresponding genomes. For each of the 157 prokaryotic genomes studied, the DNA G+C content of the 20 genes was used to determine a 'calculated' genome DNA G+C content (CGC) and this value was compared with the 'real' genome DNA G+C content (RGC). In order to select the most suitable gene for the determination of CGC values, we compared the r2 and median mol% difference between CGC and RGC as well as the sensitivity of each gene to provide CGC values for prokaryotic genomes that differ by less than 5 mol% from their RGC. The highly conserved ftsY gene (median size 1144 nucleotides), a vertically inherited member of the GTPase superfamily, showed the highest r2 value of 0.98, the smallest median mol% difference between CGC and RGC of 1.06 and a sensitivity of 100 %. Using ftsY DNA G+C content values, the CGC values of 100 genomes not included in the calculation of r2 differed by less than 5 mol% from their RGC values. These data suggest that the genomic DNA G+C content of prokaryotes may be estimated easily and reliably from the ftsY gene sequence.

Bacteria↗

rpoB gene sequence-based characterization of emerging non-tuberculous mycobacteria with descriptions of Mycobacterium bolletii sp. nov., Mycobacterium phocaicum sp. nov. and Mycobacterium aubagnense sp. nov.

Over the past 10 years, 16S rRNA gene sequencing has contributed to the establishment of more than 45 novel species of non-tuberculous mycobacteria and to the description of emerging mycobacterial infections. Cumulative experience has indicated that this molecular tool underestimates the diversity of this group and does not distinguish between all recognized mycobacterial taxa. In order to improve the recognition of emerging rapidly growing mycobacteria (RGM), rpoB gene sequencing has been developed. Our previous studies have shown that an RGM isolate is a member of a novel species if it exhibits >3 % sequence divergence in the rpoB gene from the type strains of established species. When applied to a collection of 59 clinical RGM isolates, rpoB gene sequencing revealed nine novel isolates (15.3 %) whereas only two isolates (3.4 %) were deemed to be novel by conventional 16S rRNA gene sequence analysis. A polyphasic approach, including biochemical tests, antimicrobial susceptibility analyses, hsp65, sodA and recA gene sequence analysis, DNA G+C content determination and cell-wall fatty acid composition analysis, supported the evidence that these nine isolates represent three novel species. Whereas Mycobacterium phocaicum sp. nov. (type strain N4T = CIP 108542T = CCUG 50185T) and Mycobacterium aubagnense sp. nov. (type strain U8T = CIP 108543T = CCUG 50186T; Mycobacterium mucogenicum group) were susceptible to most antibiotics, Mycobacterium bolletii sp. nov. (type strain BD(T) = CIP 108541T = CCUG 50184T; Mycobacterium chelonae-abscessus group) was resistant to the quinolones, tetracycline, macrolides and imipenem. Only M. bolletii was resistant to clarithromycin. These data illustrate that rpoB gene sequence-based identification is a powerful tool to characterize emerging RGM and mycobacterial infections and provides valuable help in differentiating RGM at both the intra- and interspecies level, thus contributing to a faster and more efficient diagnosis and epidemiological follow-up.

Anti-Bacterial Agents↗

Bacillus massiliensis sp. nov., isolated from cerebrospinal fluid.

An unidentified Gram-negative-staining, aerobic, rod-shaped, spore-forming bacterium was isolated from a sample of cerebrospinal fluid. Based on comparative analysis of 16S rRNA gene sequences and phenotypic characteristics, the novel isolate was included in the Bacillus sphaericus-like group. The isolate was closely related to Bacillus odysseyi and Bacillus silvestris, with 96.2 and 94.4 % 16S rRNA gene sequence similarity, respectively. The major fatty acid was iso-C(15 : 0) (48 %). The name Bacillus massiliensis sp. nov. is proposed for the novel isolate, with strain 4400831(T) (=CIP 108446(T)=CCUG 49529(T)) as the type strain.

Bacillaceae Infections↗

Streptococcus massiliensis sp. nov., isolated from a patient blood culture.

An unidentified strain of the viridans group of streptococci was isolated from a human blood sample. It was distinguished from all other recognized species of the Streptococcus sanguinis group by several biochemical characteristics. Phylogenetic analysis based on 16S rRNA gene sequence comparisons clustered this strain with Streptococcus ferus (mutans group) but phylogenetic analysis based on rpoB and sodA gene sequence comparisons included it in the S. sanguinis group. The isolate showed 95.4 and 95.2 % 16S rRNA gene sequence similarity to S. ferus and S. sanguinis, respectively, confirming it as belonging to a novel taxon, for which the name Streptococcus massiliensis sp. nov. is proposed. The type strain is 4401825T (=CIP 108498T=CCUG 49690T).

Bacterial Proteins↗

Rickettsia tamurae sp. nov., isolated from Amblyomma testudinarium ticks.

Rickettsia sp. strain AT-1(T) was isolated from Amblyomma testudinarium ticks in Japan in 1993. Comparative analysis of sequences obtained from 16S rRNA, gltA, ompA, ompB and sca4 gene fragments demonstrated those from AT-1(T) to be markedly different from those of other members of the spotted fever group. Using mouse serotyping, it was also observed that Rickettsia sp. strain AT-1(T) was different from other Rickettsia species with validly published names. Such genotypic and phenotypic characteristics warrant its classification as a representative of a novel species, for which the name Rickettsia tamurae sp. nov. is proposed, with the type strain AT-1(T) (=CSUR R1(T)).

Bacterial Outer Membrane Proteins↗

Rickettsia asiatica sp. nov., isolated in Japan.

The characterization of a novel Rickettsia species, first isolated from Ixodes ovatus ticks in Japan in 1993, is described for which the name Rickettsia asiatica sp. nov. is proposed. Using multilocus sequence comparison, five rickettsial isolates from I. ovatus collected at various locations in Japan were found to be identical but belonged to a novel species. Serotyping also demonstrated this rickettsia to be distinct from previously described Rickettsia species. The type strain of Rickettsia asiatica sp. nov. is IO-1(T) (=CSUR R2(T)). Following discussions held at the meeting of the International Committee on Systematics of Prokaryotes (ICSP) and its Judicial Commission (JC) in San Francisco, 2005, and in anticipation of the published minutes of the JC and ICSP, a committee consisting of the chairman of the ICSP, the chairman of the Judicial Commission of the ICSP and the Editor of the IJSEM has granted an exception in this case to the Rule governing the deposit of type material in two different collections in two different countries.

Animals↗

New insight into the diagnosis of fastidious bacterial endocarditis.

Sterile blood cultures are noted in one third of patients with infectious endocarditis. Although in half of cases this is due to previous antibiotic therapy, in the other half, the aetiology of culture-negative endocarditis is intracellular bacteria such as Coxiella burnetii or fastidious growing bacteria. Although it was previously considered that the prevalence of such organisms was identical throughout the world, recent investigations on Bartonella endocarditis clearly showed that the aetiology of culture-negative endocarditis is likely to be strongly related to epidemiology of the agent in each country. During the past decade the use of molecular techniques such as PCR with subsequent sequencing to detect or to identify bacteria in valves from patients with infectious endocarditis have considerably improved the aetiological diagnosis. This is especially true in the case of culture-negative endocarditis following earlier antibiotic therapy. However, the fact that DNA remnants of past endocarditis can be detected some time after the acute episode, when the patient has been cured, suggests that the predictive value of these techniques along with the traditional histology and culture need to be evaluated closely.

Anti-Bacterial Agents↗

Partial rpoB gene sequencing for identification of Leptospira species.

The usual target for sequence-based identification of Leptospira species is the 16S rRNA gene. However, because the 16S rRNA gene is not polymorphic enough, it is necessary to sequence a 1500 bp segment of this gene for accurate identification. Based on the alignment of previously determined rpoB of three Leptospira strains, we designed and tested a primer pair that enabled us to amplify and sequence a 600 bp segment of Leptospira rpoB. This segment was species-specific for the 16 species tested, but was unable to separate Leptospira interrogans serovars accurately. For the 11 L. interrogans serovars tested, only seven genotypes could be determined. We thus think that analysis of partial rpoB may be useful as an initial screening test for the identification of a new isolate of Leptospira and detection or identification of Leptospira in clinical or environmental samples, but not for serovar determination.

Bacterial Typing Techniques↗

Molecular characterization of resistance to macrolides in Bartonella henselae.

We selected in vitro erythromycin-resistant strains of Bartonella henselae. The mutants obtained had point mutations in domain V of 23S rRNA and/or in ribosomal protein L4. One lymph node of a patient with cat-scratch disease had such a mutation in 23S rRNA, suggesting that natural resistant strains may infect humans.

Bartonella henselae↗

Survival of environmental mycobacteria in Acanthamoeba polyphaga.

Free-living amoebae in water are hosts to many bacterial species living in such an environment. Such an association enables bacteria to select virulence factors and survive in adverse conditions. Waterborne mycobacteria (WBM) are important sources of community- and hospital-acquired outbreaks of nontuberculosis mycobacterial infections. However, the interactions between WBM and free-living amoebae in water have been demonstrated for only few Mycobacterium spp. We investigated the ability of a number (n = 26) of Mycobacterium spp. to survive in the trophozoites and cysts of Acanthamoeba polyphaga. All the species tested entered the trophozoites of A. polyphaga and survived at this location over a period of 5 days. Moreover, all Mycobacterium spp. survived inside cysts for a period of 15 days. Intracellular Mycobacterium spp. within amoeba cysts survived when exposed to free chlorine (15 mg/liter) for 24 h. These data document the interactions between free-living amoebae and the majority of waterborne Mycobacterium spp. Further studies are required to examine the effects of various germicidal agents on the survival of WBM in an aquatic environment.

Acanthamoeba↗