Objectives: Nocardia, a Gram-positive bacterium, is responsible for rare and severe infections. Accurate microbiological data are essential to guide antibiotic treatment. Our primary objective was to describe species identification and results of antimicrobial susceptibility testing (AST) for Nocardia isolates analysed over a 6-year period. Secondary objectives were to study temporal trends in species distribution and AST results. Methods: We retrospectively analysed results from Nocardia isolates sent between January 2010 and December 2015 to a French laboratory dedicated to Nocardia (Observatoire Francais des Nocardioses). Species identification was obtained by amplification and sequencing of a 600-bp fragment of the 16S rRNA gene (for all isolates) and of hsp65 (when required). AST was performed using disk diffusion. Results: We included 793 Nocardia isolates, mostly from the lungs (53.8%). The most frequent species were Nocardia farcinica (20.2%), Nocardia abscessus complex (19.9%) and Nocardia nova complex (19.5%). The proportion of N. farcinica increased significantly over time from 13% in 2010 to 27.6% in 2014. Linezolid, amikacin, trimethoprim-sulfamethoxazole, minocycline and imipenem were the most frequently identified active antibiotics with, respectively, 0% (0/734), 2.9% (21/730), 5.4% (40/734), 9.4% (69/734) and 19.5% (143/732) of isolates not susceptible. Nocardia farcinica was frequently not susceptible to cefotaxime (118/148, 79.7% of the isolates), but only about 5% of Nocardia cyriacigeorgica and N. abscessus complex isolates were not susceptible to cefotaxime. Conclusions: In this first epidemiological study of Nocardia isolated from human samples in France, N. farcinica was the species most frequently identified and its prevalence increased over time. (C) 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Les Nocardia sont des bactéries à Gram positif responsables de pathologies rares mais graves. Disposer de données microbiologiques précises (sensibilité aux antibiotiques et typage moléculaire d’espèce) sur un grand nombre de souches est indispensable afin de choisir le traitement probabiliste puis de l’adapter. Notre objectif principal était de décrire la sensibilité aux antibiotiques et l’épidémiologie moléculaire des Nocardia en France sur une large collection de souches. Notre objectif secondaire était d’étudier l’évolution au cours du temps de la résistance aux antibiotiques et de la proportion de chaque espèce et d’évaluer la répartition géographique des souches collectées. Nous avons analysé rétrospectivement les données relatives aux souches de Nocardia isolées chez l’homme durant 6 ans (2010–2015). L’identification d’espèce était obtenue par amplification et séquençage d’un fragment de 600pb du gène codant pour l’ARNr 16S (pour toutes les souches) et le gène hsp65 (pour les complexes N . abscessus et N . transvalensis ). La sensibilité aux antibiotiques était évaluée par technique de diffusion des antibiotiques en disques sur gélose Mueller-Hinton ajustée en cations. Nous avons inclus 793 souches de Nocardia , principalement isolées au niveau pulmonaire (53,8 %) ou à partir de lésions cutanées ou sous-cutanées (19,7 %). Les principales espèces étaient N . farcinica (20,2 %), N . abscessus complexe (19,9 %), N . nova complexe (19,5 %) et N . cyriacigeorgica (12,9 %). La proportion de N . farcinica augmentait significativement au cours du temps. Le linézolid, l’amikacine, le triméthoprime-sulfaméthoxazole, la minocycline, et l’imipénem étaient les antibiotiques les plus souvent actifs in vitro avec respectivement 0 %, 2,9 %, 7 %, 9,4 %, 13,8 % et 19,5 % de souches non sensibles ; ces fréquences étaient stables au cours du temps. N . farcinica était souvent non sensible aux céphalosporines de 3 e génération (∼ 80 % des souches), mais seulement 5 % des N . cyriacigeorgica et des N . abscessus complexe n’étaient pas sensibles au céfotaxime et à la ceftriaxone. Une plus grande densité de souches était observée dans le Sud-Est de la France et dans les territoires d’Outre-Mer, par rapport au Nord de la France. Dans cette première étude épidémiologique des souches de Nocardia isolées chez l’Homme en France, N . farcinica était la principale espèce identifiée et sa proportion augmentait au cours du temps.

This study aims at genetic characterization and phylogenetic relationships of Nocardia brasiliensis focusing by using housekeeping rrs, hsp65, and sodA genes. N. brasiliensis is the species responsible for 80% of cases of actinomycetoma, one form of cutaneous nocardiosis which occurs mainly in tropical regions reaching immunocompetent patients in which the disease can lead to amputation. We analyze 36 indigenous cases of N. brasiliensis that happened in France. Phylogenetic analysis targeting rrs gene showed no robustness at phylogenetic nodes level. However, the use of a concatenation of hsp65 and sodA genes showed that the tested strains surprisingly ranked in 3 well-defined genotypes. Genotypes 2 and 3 were phylogenetically closer to each other and both diverged from genotype 1 sustained by a high bootstrap of 81%. This last genotype hosts all the cases of pulmonary forms (3), the sole cerebral form, and almost all the cases of immunocompromised patients (3 out of 4). Moreover, excepting one of them, all the strains belonging to this group present a susceptibility to imipenem which is not the case in the other genotypes that rarely count among them strains being susceptible to this drug. The haplotype diversity (Hd) of hsp65 (0.927) and sodA (0.885) genes was higher than that of rrs (0.824). For this gene, we obtained 16 polymorphic sites whereas, for hsp65 and sodA genes, up to 27 and 29 were identified, respectively. This study reveals that these two genes have an important genetic discriminatory power for the evaluation of the intraspecies genetic variability of N. brasiliensis and they may be useful for identification purposes at species level. This study also reveals the possible existence of a new species harbored by genotype 1.

Currently for bacterial identification and classification the rrs gene encoding 16S rRNA is used as a reference method for the analysis of strains of the genus Nocardia. However, it does not have enough polymorphism to differentiate them at the species level. This fact makes it necessary to search for molecular targets that can provide better identification. The sodA gene (encoding the enzyme superoxide dismutase) has had good results in identifying species of other Actinomycetes. In this study the sodA gene is proposed for the identification and differentiation at the species level of the genus Nocardia. We used 41 type species of various collections; a 386 bp fragment of the sodA gene was amplified and sequenced, and a phylogenetic analysis was performed comparing the genes rrs (1171 bp), hsp65 (401 bp), secA1 (494 bp), gyrB (1195 bp) and rpoB (401 bp). The sequences were aligned using the Clustal X program. Evolutionary trees according to the neighbour-joining method were created with the programs Phylo_win and MEGA 6. The specific variability of the sodA genus of the genus Nocardia was analysed. A high phylogenetic resolution, significant genetic variability, and specificity and reliability were observed for the differentiation of the isolates at the species level. The polymorphism observed in the sodA gene sequence contains variable regions that allow the discrimination of closely related Nocardia species. The clear specificity, despite its small size, proves to be of great advantage for use in taxonomic studies and clinical diagnosis of the genus Nocardia.

Background. - Mexico has the largest number of clinical cases of actinomycetoma in North and South America. Species originally identified by less specific methods have been recently reclassified as other known species or as new species.Objective. - To assess, by 16S rRNA gene sequencing and phenotypic methods, the species distribution of 18 human clinical isolates originally identified as N. brasiliensis, some of them isolated between 1947 and 1959 in Mexico City.Methods. - Clinical isolates came from the Hospital General, "Dr. Manuel Gea Gonzalez", and Instituto Nacional de Diagnostic y Referencia Epidemiologica (INDRE) in Mexico, D.F. The strains used in this study included 15 clinical strains isolated between 1947 and 1959 that were originally identified as N. brasiliensis and three more strains obtained in 2007 identified as Nocardia spp. The isolates were identified genotypically by sequencing the 165 rRNA gene, and their phenotypic profiles were obtained with the API Coryne (R) system. Antibiotic susceptibility patterns were tested according to the protocol of the Comite de l'antibiogramme de la Societe francaise de microbiologie [4].Results. - According to 165 rRNA gene, sequencing were identified among 18 human clinical isolates as Nocardia farcinica (n = 11) and Nocardia brasiliensis (n = 7). A high number of the strains were susceptible to the majority of the antibiotics tested. The phenotypic profiles of the strains were quite uniform for N. farcinica and some variability was observed for N. brasiliensis strains.Conclusion. - N. farcinica was the most prevalent species identified. Modern methodologies should be applied in clinical laboratories to accurately identify etiological agents. (C) 2011 Elsevier Masson SAS. All rights reserved.

Objectives. - To describe the disease, its infecting agents and the increasing incidence of a pathology whose epidemiology is evolving simultaneously with improvements in identification technologies.Clinical profile, epidemiology. - Nocardiosis is a granulomatous and suppurative infection, localized or disseminated, which generally results from inhalation of the germs and, more rarely, contamination of a wound. Less and less rare, it affects mainly the immunosuppressed subjects but the observation of cases in the apparent absence of a predisposing factor is not unusual. A relationship between some strains of Nocardia and cases of Parkinson's disease is not to be excluded. The epidemiology was completely modified these last years and the causative species are in 80 to 90% of cases N. abscessus and N. cyriacigeorgica (in the past associated under the name of N. asteroides type VI), N. farcinica, N. nova and, more rarely, N. brasiliensis, N. otitidiscaviarum, to which it is necessary to add the new species recently discovered: N. veterana, N. paucivorans, etc.Identification, antibiotic susceptibility, treatment. - The precise identification is now possible only using the most modern techniques of molecular biology. Association of imipenem plus amikacine by intravenous route at the beginning of the infection can be recommended in first intention, associated or not with cotrimoxazole which wit( then be, in the majority of cases, used alone per os during several months. The primary cutaneous forms require a treatment of at [east 3 months; the pulmonary forms classically impose a 6-month treatment; the disseminated forms and the patients presenting an immunosupression require treatments of at least 12 months, which can sometimes be prolonged for life. (c) 2008 Elsevier Masson SAS. Tous droits reserves.

Las bacterias del genero Nocardia son actinomicetos aerobios cuyo habitat es el suelo y la materia organica en descomposicion. La infeccion natural por especies de este genero ha sido reportada en humanos y animales, sin embargo, la infeccion natural en ratones no ha sido descrita. En este estudio se demuestra por primera vez el aislamiento de la especie recientemente descrita, Nocardia cyriacigeorgica , en pulmones de ratones Balb/c. El estudio microbiologico de las muestras de pulmon reporto la presencia de filamentos grampositivos ramificados fragmentados en forma cocobacilares. Los estudios de PCR demostraron que la especie aislada fue N. cyriacigeorgica Se evidencia en este trabajo que N. cyriacigeorgica es potencialmente patogena en ratones Balb/c.

Human infection caused by Phaeoacremonium parasiticum is increasingly being reported. However, only a few case reports show its role as a soil-related contaminant of kidney recipients. We report here a case of a subcutaneous infection by P. parasiticum in a transplanted man. After 4 years, he reported a nonpainful nodule on his forefinger. It was surgically excised. Histological examination revealed dense fibrous connective tissue showing an extensive granulomatous reaction, including the presence of a wooden sliver, and hyaline, branched, and septate hyphae. Six months later, the patient relapsed and underwent large excision. Culture was positive for P. parasiticum. No antifungal drug was administered. Surgical excision was successful.

Nous rapportons un cas d′endophtalmie rarissime à Lasiodiplodia theobromae. Un homme consulta pour une douleur oculaire après traumatisme par branche d′arbre. L′acuité visuelle était chiffrée à « voit bouger la main ». À l′examen, un abcès de cornée ainsi qu′une inflammation de chambre antérieure étaient notés. Devant cet aspect, un traitement antibiotique local et général fut instauré. L′examen direct du grattage cornéen mit en évidence la présence de filaments mycéliens. Un traitement par amphotéricine B collyre et itraconazole per os fut prescrit. La culture permit l′isolement d′un champignon filamenteux. L′identification de Lasiodiplodia theobromae sera réalisée par une technique de biologie moléculaire. Malgré le traitement, le tableau clinique évolua vers une panophtalmie gravissime, pour laquelle le patient subit une éviscération. L′intérêt de cette observation réside en une mise au point sur Lasiodiplodia theobromae, et dans le rappel des principaux facteurs de risque et éléments cliniques retrouvés dans les kératomycoses. We report a case of endophthalmitis caused by the fungus Lasiodiplodia theobromae. A 68-year-old man was referred to the hospital for right ocular pain since experiencing right ocular branch trauma 2 weeks before. The best-corrected acuity was limited to hand motion. Slit-lamp examination showed a large corneal abscess and an anterior chamber reaction. The patient underwent systemic and local antibiotic therapy, and corneal scraping for microbiological diagnosis. Sabouraud-chloramphenicol-gentamicin agar disclosed filamentous fungus, which was treated with oral itraconazole and topic amphotericin B. Molecular biology revealed Lasiodiplodia theobromae. Despite antimycotic drugs, severe panophthalmia occurred very quickly and led to evisceration. This case report describes Lasiodiplodia theobromae as the cause of keratomycosis and discusses risk factors and clinical features of fungal keratitis in order to improve prognosis by earlier treatment.

We report a case of endophthalmitis caused by the fungus Lasiodiplodia theobromae. A 68-year-old man was referred to the hospital for right ocular pain since experiencing right ocular branch trauma 2 weeks before. The best-corrected acuity was limited to hand motion. Slit-lamp examination showed a large corneal abscess and an anterior chamber reaction. The patient underwent systemic and local antibiotic therapy, and corneal scraping for microbiological diagnosis. Sabouraud-chloramphenicol-gentamicin agar disclosed filamentous fungus, which was treated with oral itraconazole and topic amphotericin B. Molecular biology revealed Lasiodiplodia theobromae. Despite antimycotic drugs, severe panophthalmia occurred very quickly and led to evisceration. This case report describes Lasiodiplodia theobromae as the cause of keratomycosis and discusses risk factors and clinical features of fungal keratitis in order to improve prognosis by earlier treatment.

Twenty-five strains of actinomycetes were isolated from samples of water, soil and tree barks collected at two sites located in the north-east of Algeria. Antimicrobial activity was tested using the agar cylinder method against three Gram-positive bacteria, three Gram-negative bacteria, three yeasts and three filamentous fungi. Among the 25 isolates 14 (56%) strains showed an activity against at least one of the test-bacteria studied and two (8%) showed antifungal activity. Ninety-three percent of the active strains were identified by the universal PCR as belonging to the Streptomyces genus and 7% to the Actinomadura genus. (c) 2005 Elsevier SAS. All rights reserved.

Dans les pays en développement où les antifongiques systémiques sont souvent indisponibles, le traitement d’une infection à champignon filamenteux comme les Fusarium s’avère parfois très difficile. Nous rapportons le cas d’une kératomycose à Fusarium oxysporum traitée par la povidone iodée en collyre et le fluconazole per os. Le diagnostic d’abcès de la cornée était retenu après examen ophtalmologique chez un homme de 28 ans sans antécédent ophtalmologique, adressé à la clinique d’ophtalmologie du CHU Le Dantec de Dakar pour un œil gauche rouge douloureux avec une baisse de l’acuité visuelle évoluant depuis 15 jours. Le patient n’avait pas reçu de corps étranger dans l’œil. Des prélèvements par grattage cornéen ont été réalisés pour analyse microbiologique et le patient a été hospitalisé et mis sous un traitement à base de collyres renforcés (céftriaxone + gentamicine). Le diagnostic mycologique a révélé la présence d’un champignon filamenteux saprophyte de l’environnement : F. oxysporum, ce qui a motivé un remplacement du traitement initial par un collyre de préparation magistrale à base de solution de povidone iodée à 1 % en attendant l’acquisition de l’amphotéricine B qui était indisponible sur place. Mais devant l’indisponibilité de cette molécule et la menace de la perte visuelle, un traitement à base de fluconazole per os avait été adjoint au traitement local par le collyre de povidone iodée. L’évolution a été favorable avec une guérison de l’abcès et une acuité visuelle chiffrée à VLDB (1/200e). Par ailleurs nous notions des séquelles à types de pannus et de taie. Une vulgarisation des antifongiques topiques d’efficacité démontrée dans les pays en développement serait nécessaire pour optimiser leur traitement.In developing countries where systemic antifungal are often unavailable, treatment of filamentous fungi infection as Fusarium is sometimes very difficult to treat. We report the case of a keratomycosis due to Fusarium oxysporum treated by povidone iodine eye drops and oral fluconazole. The diagnosis of abscess in the cornea was retained after ophthalmological examination for a 28-year-old man with no previous ophthalmological disease, addressed to the Ophthalmological clinic at the University Hospital Le Dantec in Dakar for a left painful red eye with decreased visual acuity lasting for 15 days. The patient did not receive any foreign body into the eye. Samples by corneal scraping were made for microbiological analysis and the patient was hospitalized and treated with a reinforced eye drops based treatment (ceftriaxone + gentamicin). The mycological diagnosis revealed the presence of a mold: F. oxysporum, which motivated the replacement of the initial treatment by eye drops containing iodized povidone solution at 1 % because of the amphotericin B unavailability. Due to the threat of visual loss, oral fluconazole was added to the local treatment with eye drops povidone iodine. The outcome was favorable with a healing abscess and visual acuity amounted to 1/200th. Furthermore, we noted sequels such as pannus and pillowcase. The vulgarization of efficient topical antifungal in developing countries would be necessary to optimize fungal infection treatment.