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Biomedical subjects

P Boiron

Publications and source records attributed to P Boiron.

At least 37 records · Page 2Linked to original sources

Biochemical-genetic analysis and distribution of FAR-1, a class A beta-lactamase from Nocardia farcinica.

From genomic DNA of the clinical isolate Nocardia farcinica VIC, a 1. 6-kb Sau3AI fragment was cloned and expressed in Escherichia coli JM109. The recombinant strain expressed a beta-lactamase (pI, 4.6), FAR-1, which conferred high levels of resistance to amoxicillin, piperacillin, ticarcillin, and cephalothin. The hydrolysis constants (kcat, Km, Ki, and 50% inhibitory concentration) confirmed the MIC results and showed that FAR-1 activity is inhibited by clavulanic acid and at a low level by tazobactam and sulbactam. Moreover, FAR-1 beta-lactamase hydrolyzes aztreonam (at a low level) without significant activity against ceftazidime, cefotaxime and imipenem. FAR-1 mature protein of molecular mass ca 32 kDa, has less than 60% amino acid identity with any other class A beta-lactamases, being most closely related to PEN-A from Burkholderia cepacia (52%). A blaFAR-1-like gene was found in all studied N. farcinica strains, underlining the constitutive origin of this gene.

Amino Acid Sequence↗

Rapid identification of clinically relevant Nocardia species to genus level by 16S rRNA gene PCR.

Two regions of the gene coding for 16S rRNA in Nocardia species were selected as genus-specific primer sequences for a PCR assay. The PCR protocol was tested with 60 strains of clinically relevant Nocardia isolates and type strains. It gave positive results for all strains tested. Conversely, the PCR assay was negative for all tested species belonging to the most closely related genera, including Dietzia, Gordona, Mycobacterium, Rhodococcus, Streptomyces, and Tsukamurella. Besides, unlike the latter group of isolates, all Nocardia strains exhibited one MlnI recognition site but no SacI restriction site. This assay offers a specific and rapid alternative to chemotaxonomic methods for the identification of Nocardia spp. isolated from pathogenic samples.

Bacteriological Techniques↗

Localized subcutaneous Nocardia farcinica abscess in a woman with overlap syndrome between systemic scleroderma and polymyositis.

Nocardia farcinica is an increasingly common cause of human infection in Europe. We observed an isolated nodular subcutaneous abscess due to N. farcinica in the forearm of a young patient with overlap syndrome between systemic sclerosis and polymyositis. Despite in vitro resistance to trimethoprim-sulfamethoxazole, this oral combination completely resolved the lesion within 3 weeks and was maintained for 6 months. The spectrum of cutaneous nocardiosis in the immunocompromised host is discussed.

Abscess↗

Nocardia, nocardiosis and mycetoma.

The recent emergence of invasive infections due to Nocardia spp., including nosocomial outbreak, is now evident. Newer molecular diagnostic and typing methods are developed. Although sulfonamide-based therapy is generally effective, optimal treatment may be guided by antimicrobial susceptibility testing of isolates. The improved classification of nocardiae and other related genera such as actinomadurae, using the 16S ribosomal RNA sequencing, provide a sound basis for improved diagnostic methods for the identification of members of clinically significant species. The commonest cause of eumycetoma in Sudan is Madurella mycetomatis, and Streptomyces somaliensis and Actinomadura madurae for actinomycetoma. The humoral immunity response in actinomycetoma patients and in experimental mice was measured and significant titre of anti-P24 antibody was demonstrated.

Actinomycetales↗

Nocardia otitidiscaviarum infection of a traumatic skin wound.

A case of primary Nocardia otitidiscaviarum infection of a skin wound over an open fracture in a previously healthy adult who suffered multiple trauma in a car accident is reported. The organism was identified in cultures of pus specimens from the infected wound. The case demonstrated the difficulties of testing susceptibility of nocardiae in vitro and the necessity of prolonged antibiotic treatment. The prevalence of nocardial infections is underestimated, highlighting the need for adequate documentation of such infections and compilation of the information by public health authorities.

Adult↗

Nocardia asteroides in the soil of Kuwait.

A pilot study was undertaken to determine the occurrence and distribution of pathogenic nocardiae in Kuwaiti soil. A total of 102 soil samples collected from two localities were investigated by the paraffin bait technique. Nocardia asteroides was the only species isolated from 42 (41%) soil samples. None of the isolates fulfilled the criteria required for identification of N. farcinica or N. nova. Thirty one (73.8%) isolates showed equivalent growth at 45 degrees C and 35 degrees C, 17 (40.4%) isolates utilized acetamide for carbon and nitrogen requirements and 3 (7.1%) isolates showed delayed arylsulphatase activity. Only a solitary isolate was resistant to cefamandole. Soil samples originating from the Kuwait University Campus, Shuwaikh, which were rich in humus/organic matter, were more productive for N. asteroides (67%) than the samples which were devoid of it but were mixed with crude oil (39%). Sand samples that lacked organic matter and crude oil samples were least productive of N. asteroides. These preliminary findings do not suggest that massive oil contamination of soil in the Ahmadi oil field area during the Gulf war promoted the natural occurrence of N. asteroides. However, isolation of N. asteroides in as many as 41% of the soil sample is a significant observation warranting further epidemiologic studies including its possible role in the operation desert storm sickness syndrome. This is the first report on the natural occurrence of N. asteroides in Kuwait.

Fuel Oils↗

Transmission of nocardiosis and molecular typing of Nocardia species: a short review.

Nocardia species are ubiquitous in the environment and may be found in the soil. They are generally responsible for sporadic pulmonary diseases acquired by inhalation of spores, with secondary localizations in the central nervous system and subcutaneous tissues. There is no absolute evidence for person to person transmission. Presumptive outbreaks of nocardiosis were observed in immunocompromised patients, more frequently in kidney transplant patients than in cardiac transplant patients. Nocardia spp., being present in dust particles, closure and disinfection of the transplantation unit with formaldehyde arrested the sequence of cases of nocardiosis. The original sources of the Nocardia sp. remain doubtful. Other possible sources of contamination are other patients, medical staff and the hospital environment. The first studies of Nocardia spp. typing were based on the detection of extracellular antigens, on the susceptibility of actinomycete strains to killer yeasts, and on the biochemical profiles with fluorogenic substrate. The use of molecular typing techniques have given very promising results. Analysis of plasmid profiles is an interesting way to compare the identity of isolates, although the reliability of this method depends of the presence of plasmids in the isolates. Other typing methods, including analysis of restriction length fragment polymorphism of total DNA, ribosomal DNA fingerprinting, require further investigations to evaluate their discriminating power or to be easily interpretable, whereas a random amplified polymorphic DNA (RAPD) assay was successful for epidemiological purposes. Progress in epidemiological analysis of cases of nocardiosis will be consistent when an improved diagnosis of this infection (molecular and serological diagnosis) will be available, when the genetic diversity of Nocardia spp. isolates will be better known, and when molecular typing, that hold promise in complementing investigations of outbreak of these infections, will be systematically performed when an abnormal increase of cases of nocardiosis in a population with risk factors is observed.

Disease Outbreaks↗

Severe keratitis due to Nocardia farcinica.

Keratitis due to Nocardia farcinica occurred in a 49-year-old female after inappropriate cleaning of her semipermeable rigid contact lenses with basin-stored water during a holiday in France. N. farcinica was differentiated from Nocardia asteroides by its growth at 45 degrees C, acid production from rhamnose, its opacification of Middlebrook 7H10 agar, and its marked degree of resistance to all cephalosporins, aminoglycosides, tetracyclins, macrolides, and trimethoprim-sulfamethoxazole. To the best of our knowledge, this is the first reported case of human N. farcinica keratitis, confirming that this microorganism can be responsible for serious human disease.

Contact Lenses↗

Molecular study of persistence of Nocardia asteroides and Nocardia otitidiscaviarum strains in patients with long-term nocardiosis.

Three Nocardia asteroides and five Nocardia otitidiscaviarum isolates recovered from three patients with long-term nocardiosis were compared by random amplification of polymorphic DNA fingerprinting, antibiotic susceptibility testing, and enzymatic characterization. Results obtained with primer DKU49 (P. Palittapongarnpim, S. Chomyc, A. Fanning, and D. Kunimoto, J. Infect. Dis. 167:975-978, 1993) provide evidence that patient A was infected by two N. asteroides strains during a single episode of nocardiosis and that patients B and C remained infected by the same strain, respectively. Resistance to minocycline that was present in the first isolate recovered from patient B reverted to intermediate resistance in the second isolate and reverted to susceptibility in the third isolate. Resistance to penicillin G and beta-lactams was acquired by the second isolate obtained from patient C.

DNA Fingerprinting↗

Nocardia pseudobrasiliensis sp. nov., a new species of Nocardia which groups bacterial strains previously identified as Nocardia brasiliensis and associated with invasive diseases.

We studied five strains of a new Nocardia taxon recently identified among Nocardia brasiliensis strains associated with invasive diseases (R. J. Wallace, Jr., B. A. Brown, Z. Blacklock, R. Ulrich, K. Jost, J. M. Brown, M. M. McNeil, G. Onyi, V. A. Steingrube, and J. Gibson, J. Clin. Microbiol. 33:1528-1533, 1995) to determine their taxonomic status. Several characteristics of these organisms, including the presence of chemotype IV cell walls, nocardomycolic acids, a predominant menaquinone similar to that of Nocardia asteroides ATCC 19247T (T = type strain), and G+C contents ranging from 67 to 68 mol%, are characteristics of the genus Nocardia. Phylogenies based on small-subunit ribosomal DNA sequences clearly confirmed that all five strains belong to the genus Nocardia and occur on a single branch that is clearly distinct from N. brasiliensis. This branch forms a clade with Nocardia vaccinii, Nocardia nova, Nocardia otitidiscaviarum, and Nocardia seriolae. The five new strains exhibited high levels of DNA relatedness with each other, as determined by DNA-DNA hybridization experiments (S1 nuclease procedure), but not with N. brasiliensis strains or with strains of the four phylogenetically related Nocardia species mentioned above. The five new strains differ from N. brasiliensis in the following characteristics: mycolic acid pattern, decomposition of adenine, nitrate reduction, and antimicrobial agent susceptibilities. Therefore, we propose that these strains belong to a new species, Nocardia pseudobrasiliensis. The type strain is strain ATCC 51512, which was isolated from a leg abscess on a patient suffering from ulcerative colitis.

DNA, Bacterial↗

Occurrence of plasmids in pathogenic strains of Nocardia.

The purpose of the present study was to determine the relative distribution of plasmids in 87 clinical isolates of Nocardia, belonging to the five major pathogenic species. A correlation between plasmid content and the site of infection within the host, resistance to antibiotics and enzymic profiles was also investigated. The plasmid extraction procedure of Kado and Liu was used. Electrophoretic analysis revealed one-to-four plasmid bands, ranging in size from <8 to >50 kb, in 27 strains (31%). Based on the number of isolates tested, the incidence of plasmid-bearing strains was significantly higher among N. farcinica than N. asteroides strains. Within N. farcinica, the incidence of plasmids was higher among strains isolated in the Paris area than in strains isolated elsewhere, such as in the French provinces or outside France. A statistically significant correlation was demonstrated between the cutaneous localisation of infections and the incidence of plasmid-bearing strains. The presence of plasmids in nocardiae could not be associated with specific phenotypic traits such as resistance to antibiotics or enzymic activity. The fact that the majority of Nocardia clinical isolates (60 of 87) did not contain plasmids suggests that plasmids are not involved directly in virulence and that there is no selective pressure for plasmid acquisition.

Anti-Bacterial Agents↗

Molecular study of nosocomial nocardiosis outbreak involving heart transplant recipients.

Randomly amplified polymorphic DNA analysis and rRNA gene restriction patterns (ribotyping) were compared as methods of investigating a nosocomial outbreak of nocardiosis involving three heart transplant recipients. No clear distinctions between three clinically related isolates and four unrelated strains were obtained by ribotyping. On the contrary, randomly amplified polymorphic DNA analysis with two selected primers, primers 2650 and DKU49, showed one pattern for the three related isolates and four patterns for the unrelated strains.

Adult↗

Ribotyping: a tool for taxonomy and identification of the Nocardia asteroides complex species.

Ribotyping has been evaluated as a taxonomic tool for studying bacteria belonging to the Nocardia asteroides complex. The probe used was obtained by PCR from DNA extracted from the type strain Nocardia asteroides ATCC 19247, a sequence that codes partly for 16S rRNA. Interpretation of hybridization after EcoRI restriction of total DNA of 21 strains of the N. asteroides complex allowed for the identification of only four different ribotypes: two related to N. asteroides sensu stricto, one related to N. farcinica and one related to N. nova. These results confirm the classification established by Tsukamura in 1982 (M. Tsukamura, Microbiol. Immunol. 26:1101-1119, 1982) as well as the heterogeneity of N. asteroides sensu stricto species. Ribotyping may also provide a major identification technique for the species belonging to the N. asteroides complex.

Bacterial Typing Techniques↗

Studies on siderophore production and effect of iron deprivation on the outer membrane proteins of Madurella mycetomatis.

The purpose of this investigation was to determine whether Madurella mycetomatis, the most frequent agent of eumycotic mycetomas, produces siderophores and synthesizes new outer membrane proteins under iron-starvation conditions. Siderophore production, only of the hydroxamate type, was demonstrated in all nine strains tested. It was regulated by extracellular iron concentrations. Under iron-restricted conditions, M. mycetomatis expressed various outer membrane iron-regulated proteins, particularly of 24-kilodalton, that may participate in iron metabolism.

Bacterial Outer Membrane Proteins↗

Phylogeny of the genus Corynebacterium deduced from analyses of small-subunit ribosomal DNA sequences.

We determined almost complete small-subunit ribosomal DNA sequences of 50 reference strains belonging to the genera Corynebacterium, Rhodococcus, and Gordona and compared these sequences with previously published sequences. Three phylogenetic methods (the neighbor-joining, maximum-likelihood, and maximum-parsimony methods), as well as a bootstrap analysis, were used to assess the robustness of each topology which we obtained. The results of comparative phylogenetic analyses confirmed that the genera Corynebacterium, Dietzia, Gordona, Mycobacterium, Nocardia, Tsukamurella, and Turicella form a monophyletic taxon within the phylum containing the high-G+C-content gram-positive bacteria. The genus Corynebacterium appeared to be a monophyletic unit whose members could be divided into four major clusters. The validity of the genus Turicella is doubtful since members of this genus clearly belong to the genus Corynebacterium. The variability of chemotaxonomic characteristics within the genus Corynebacterium suggests that small-subunit ribosomal DNA sequence analysis is probably the most straightforward method for confirming that a bacterium belongs to this genus.

Base Sequence↗

Use of yeast killer system to identify species of the Nocardia asteroides complex.

We evaluated the ability of the yeast killer system to differentiate members belonging to the Nocardia asteroides complex (Nocardia asteroides, Nocardia farcinica, and Nocardia nova). Nocardia strains were selected randomly from clinical isolates. Type strains of each Nocardia species and recognized killer yeasts were taken from different collections. A clear area of inhibition surrounding the yeast cells demonstrated a positive killer effect on Nocardia spp. Two yeast strains, Pichia mrakii (K9) and Pichia lynferdii (K76), showed different killer activities against each Nocardia species. The group N. asteroides was identified as K9+ K76-, the group N. farcinica was identified as K9- K76-, and the group N. nova was identified as K9+ K76+. The three killer identifications correlated with specific taxonomic groups determined by using classical methods. The yeast killer system may be a useful means for identifying organisms within the N. asteroides complex.

Bacterial Typing Techniques↗