Physiological typing of Pseudallescheria and Scedosporium strains using Taxa Profile, a semi-automated, 384-well microtitre system.

During the last few decades, Pseudallescheria and Scedosporium infections in humans are noted with increasing frequency. Multi-drug resistance commonly occurring in this species complex interferes with adequate therapy. Rapid and correct identification of clinical isolates is of paramount significance for optimal treatment in the early stages of infection, while strain typing is necessary for epidemiological purposes. In view of the development of physiological diagnostic parameters, 570 physiological reactions were evaluated using the Taxa Profile Micronaut system, a semi-automatic, computer-assisted, 384-well microtitre platform. Thirty two strains of the Pseudallescheria and Scedosporium complex were analysed after molecular verification of correct species attribution. Of the compounds tested, 254 proved to be polymorphic. Cluster analysis was performed with the Micronaut profile software, which is linked to the ntsypc® program. The systemic opportunist S. prolificans was unambiguously separated from the remaining species. Within the P. boydii/P. apiosperma complex differentiation was noted at the level of individual strains, but no unambiguous parameters for species recognition were revealed.

Dietzia cinnamea sp. nov., a novel species isolated from a perianal swab of a patient with a bone marrow transplant.

The taxonomic status of a bacterium isolated from a perianal swab of a patient with a bone marrow transplant was characterized using a polyphasic taxonomic approach. Chemotaxonomic investigations revealed the presence of cell wall chemotype IV, short chain mycolic acids that co-migrated with those extracted from members of the genus Dietzia, and a dihydrogenated menaquinone with eight isoprene units as the predominant menaquinone. Generic assignment was confirmed by 16S rRNA gene sequencing. Comparative analysis of the 16S rRNA gene sequence showed that this isolate constitutes a distinct phyletic line within the genus Dietzia, displaying 97.5-98.7% sequence similarity with Dietzia species with validly published names. The isolate could be distinguished from the type strain of Dietzia maris (1.6% sequence divergence) and other species of the genus Dietzia by DNA-DNA hybridization, as well as by using a set of biochemical tests. Genotypic and phenotypic data show that the strain merits classification as a novel species of the genus Dietzia for which the name Dietzia cinnamea sp. nov. is proposed; the type strain is IMMIB RIV-399T (= DSM 44904T = CCUG 50875T).

Reclassification of Nocardia corynebacterioides Serrano et al. 1972 (Approved Lists 1980) as Rhodococcus corynebacterioides comb. nov.

The type strain of Nocardia corynebacterioides was the subject of a polyphasic taxonomic study. The 16S rRNA gene sequence was aligned with the sequences of representatives of the genera Corynebacterium, Dietzia, Gordonia, Mycobacterium, Nocardia, Rhodococcus, Skermania, Tsukamurella and Williamsia, and phylogenetic trees were constructed by using maximum-parsimony, maximum-likelihood and neighbour-joining methods. It was evident from the phylogenetic analysis that N. corynebacterioides represents a distinct phyletic line within the genus Rhodococcus. Menaquinone analysis showed that the organism contained dihydrogenated menaquinone with eight isoprene units, MK-8(H(2)), as the major isoprenologue. The genealogical evidence, together with chemotaxonomic and phenotypic data from this and previous studies, indicates that N. corynebacterioides DSM 20151(T) (= CIP 104510(T)) should be reclassified in the genus Rhodococcus as Rhodococcus corynebacterioides comb. nov.

Wound infection due to Absidia corymbifera and Candida albicans with fatal outcome.

A case of a mixed infection due to Candida albicans and the zygomycete Absidia corymbifera in a 38-year-old, previously healthy, Caucasian male is presented. The infection developed following serial rib fractures, and ruptures of kidney, liver and biliary tract as well as a pancreatic contusion resulting from a traffic accident. During intensive care treatment the patient underwent several surgical procedures but subsequently experienced multi-organ failure and sepsis. Some weeks later, fungal growth was observed macroscopically on the patient's skin and wounds. From wound swabs C. albicans and A. corymbifera were grown. Histopathology of abdominal tissue yielded pseudohyphae and coenocytic hyphae. Although surgical debridement and antifungal treatment with amphotericin B and 5-flucytosine were started immediately, the patient died in therapy-refractory septic multi-organ failure.

Isolation of fungi, especially Exophiala dermatitidis, in patients suffering from cystic fibrosis. A prospective study.

BACKGROUND: Patients with cystic fibrosis (CF) are at an increased risk of pulmonary colonisation by opportunistic micro-organisms. Using specialised methods, the black yeast Exophiala dermatitidis could consistently be cultured from CF patients. Isolation rates from sputum samples ranged between 1.8 and 15.7%. Occasionally, infection could be recognised. OBJECTIVES: This study aimed at investigating the isolation rates of E. dermatitidis in samples taken from CF patients at the University of Bonn, Germany. METHODS: Altogether, 439 respiratory specimens taken from 81 CF patients were screened for the occurrence of E. dermatitidis over a period of 18 months. For the selective isolation of this fungus erythritol-chloramphenicol agar (ECA) produced in house was applied. RESULTS: The isolation rate of E. dermatitidis was 1.1% from all specimens, 1.6% from all sputum samples and 6.2% in all patients examined. CONCLUSIONS: Prior to the introduction of ECA, E. dermatitidis had never been isolated in our laboratory, either from CF, or from any other patient. During this study, E. dermatitidis was found to colonise the respiratory tract of some CF patients. The use of additional selective culture media is necessary for the recognition of uncommon fungi, e.g. E. dermatitidis, in CF patients.

Fatal pulmonary scedosporiosis.

We report on a case of scedosporiosis in a 72-year-old German woman. Her disease started with a purulent ulceration of unknown course at her left foot. Soon after onset of oral antibacterial therapy she needed in-hospital treatment because of an acute pneumonia. The infection progressed despite the application of different antibiotics. Microscopic examination of tracheal fluid revealed fungal hyphae and therefore treatment with itraconazole was initiated. However, the patient developed renal failure, required mechanical ventilation and finally died in treatment-resistant septic shock. Post-mortem Scedosporium apiospermum was cultured from lung tissue taken during autopsy. This is the fourth case of human infection caused by Scedosporium species diagnosed in our laboratory during the last 4 years.

Nocardia puris sp. nov.

A bacterial isolate (IMMIB R-145(T)) from a human abscess was subjected to a polyphasic taxonomic study. Chemotaxonomic investigations revealed the isolate to have cell wall chemotype IV and mycolic acids consistent with characteristics of the genus NOCARDIA: Comparative 16S rRNA gene sequencing showed that the isolate constituted a distinct subline within the genus Nocardia, displaying greater than 1.7 % sequence divergence with established species. However, a DNA-DNA hybridization study demonstrated unambiguously that the isolate was genealogically distinct from closely related species, namely, Nocardia abscessus, Nocardia cyriacigeorgica and Nocardia farcinica, with which it showed high levels of 16S rDNA sequence similarity (97.8, 97.9 and 98.3 %, respectively). Based on phenotypic and phylogenetic data, it is proposed that this isolate be classified as a new species of the genus Nocardia, for which the name Nocardia puris (type strain IMMIB R-145(T)=DSM 44599(T)=NRRL B-24204(T)) is proposed.

Human cervicofacial actinomycoses: microbiological data for 1997 cases.

Actinomycoses are sporadically occurring endogenous polymicrobial inflammatory processes, in which fermentative actinomycetes of the genera Actinomyces, Propionibacterium, or Bifidobacterium act as the principal pathogens. Difficulties in diagnosing the disease in a timely and reliable fashion have led clinicians and microbiologists to grossly underestimate its medical importance. Therefore, we evaluated microbiological and selected clinical data derived from 1997 culture-positive cases of human cervicofacial actinomycoses examined in our laboratories during 1972-1999. The causative actinomycetes belonged to at least 9 different species, among which Actinomyces israelii and Actinomyces gerencseriae predominated. The well-known predisposition of male patients to acquire the disease varied with age and appeared to be especially pronounced in patients aged 20-60 years, the highest incidence being found in female patients aged 11-40 years and in male patients aged 21-50 years. The relevant procedures necessary for diagnosing human actinomycoses reliably, as well as details of their complex etiology, are discussed.

Mycetoma due to Pseudallescheria boydii and co-isolation of Nocardia abscessus in a patient injured in road accident.

We report the case of a patient who developed a mycetoma after experiencing a road accident. From surgical biopsies Pseudallescheria boydii was isolated. Subsequently, after the infection had been treated with itraconazole, a Gram-positive bacterium, identified as the newly described species Nocardia abscessus, was cultured from wound fluids.

Case Report. Maxillary sinus infection due to Emericella nidulans.

Fungal infections of the maxillary sinus are frequently caused by Aspergillus species, particularly A. fumigatus. In otherwise healthy persons there is an association with overfilling of dental root canals, when zinc-containing filling materials were used. Below, a maxillary sinus aspergilloma is reported in a young immunocompetent female patient caused by Aspergillus (Emericella) nidulans.

Isolation of Nocardia paucivorans from the cerebrospinal fluid of a patient with relapse of cerebral nocardiosis.

Nocardia paucivorans represents a new species of the genus Nocardia that has recently been isolated from bronchial secretions of a patient with chronic lung disease. Here, we report on the course of a disseminated infection caused by this species: i.e., cerebral and subsequent meningeal manifestations, isolation from the cerebrospinal fluid, and in vitro susceptibility to various antimicrobial agents.

A case of imported paracoccidioidomycosis in a German legionnaire.

We report on a case of the chronic form of paracoccidioidomycosis with swelling and ulcerations of the mouth in a German legionnaire who also suffered from a chronic bronchitis. The patient had worked for many years in Brazil, an area endemic for the disease. Infection due to Paracoccidioides brasiliensis was diagnosed in Germany, more than 10 years after the patient's return. Diagnosis was established by the presence of yeast cells with multipolar budding in the tissue of the oral lesion. Furthermore, the fungus was grown in a liquid Leishmania culture medium. Identification of the fungus was based on morphology and genetic sequencing. Furthermore, IgG antibodies against a 43-kDa antigen of P. brasiliensis were detected in a western blot. After itraconazole therapy (400 mg day(-1)) for 4 weeks, the lesions had disappeared almost completely, but the therapy was continued for further 5 months to avoid relapse of the infection.

Typing and antimicrobial susceptibilities of methicillin resistant Staphylococcus aureus (MRSA) strains isolated in a hospital in Korea.

Methicillin-resistant Staphylococcus aureus (MRSA) strains may cause serious nosocomial infections, including pneumonia and septicemia. The rate of methicillin-resistance among S. aureus isolates in Korea is over 50%. In this study, 90 MRSA isolates from Kyung Hee University Hospital were characterized employing bacteriophage typing, pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing. Eighty percent of the strains could be phage-typed. The largest group or 40% of the strains belonged to lyso group III, followed by 32% of the isolates which produced a reaction with regional additional phages. Phage type 83A was most frequently encountered, followed by phage type D11. PFGE patterns confirmed the presence of two major clusters, which comprise the isolates belonging to lyso group III and the strains that were typable with regional additional phages. The latter group also contained a number of strains that were nontypable with bacteriophages. The resistance rates to ciprofloxacin, erythromycin, tetracycline, gentamicin and clindamycin were over 94%. Strains with intermediate resistance to vancomycin strains or resistance to mupirocin were not found. In conclusion, this study demonstrates that the results of phage typing are confirmed and supplemented by PFGE data.

Case report. Fungaemia due to Penicillium piceum, a member of the Penicillium marneffei complex.

Due to the inability of most Penicillium species to grow at 37 degrees C, systemic non-marneffei infections are very rare in the human host. We describe a case of fungemia due to Penicillium piceum in a female patient, who died a few days after repeated isolation of this fungus from blood cultures. The species is a member of the section Biverticillata of Penicillium, as was confirmed by rDNA Internal Transcribed Spacer (ITS) sequence data, and hence may share virulence factors with P. marneffei.

Nocardia abscessus sp. nov.

Chemotaxonomic and 16S rRNA gene sequence analyses of four bacterial strains isolated from clinical material clearly demonstrated that these bacteria belong to the genus Nocardia. DNA-DNA hybridization data as well as the physiological characteristics of the isolates indicated that they are closely related and belong to a single species that differs from previously described members of the genus. The name Nocardia abscessus sp. nov. is proposed for these organisms represented by strain IMMIB D-1592T (= DSM 44432T). Strain IMMIB D-1592T exhibits 56.8 and 60.0% DNA-DNA relatedness to Nocardia asteroides ATCC 19247T and Nocardia paucivorans DSM 44386T, respectively.

Nocardia paucivorans sp. nov.

Chemotaxonomic and 16S rDNA sequence analyses of an isolate from the sputa and bronchial secretions of a patient with chronic lung disease clearly demonstrated that it belongs to the genus Nocardia. DNA-DNA hybridization data, as well as the biochemical characteristics of the isolate, indicate that it belongs to a new species that differs from previously described members of the genus Nocardia. The name Nocardia paucivorans sp. nov. is proposed for this isolate and is represented by strain IMMIB D-1632T (= DSM 44386T).

[Scedosporiosis of the brain with fatal outcome after traumatizatio of the foot. case report]. / Scedosporiose des Gehirns mit letalem Ausgang nach Fussverletzung. Fallbericht.

A 33 year old German man suffered from a wound of his foot after an accident. He developed an osteomyelitis. From wound probes Scedosporium apiospermum could be isolated for several times. Some weeks later a cerebral lesion could be diagnosed. The lesion was extirpated and again S. apiospermum could be cultured from the cerebral probes. A few weeks later the patient died and some new cerebral lesions could be diagnosed by CT-scan.

The cell wall porin of the gram-positive bacterium Nocardia asteroides forms cation-selective channels that exhibit asymmetric voltage dependence.

Detergent-solubilized cell wall extracts of the gram-positive, strictly aerobic bacterium Nocardia asteroides contain channel-forming activity as judged from reconstitution experiments using lipid bilayer membranes. The cell wall porin was identified as a protein with an apparent molecular mass of about 84 kDa based on SDS-PAGE. The porin was purified to homogeneity using preparative SDS-PAGE. The 84-kDa protein was no longer observed after heating in SDS buffer. The presumed dissociation products were not observed on SDS-polyacrylamide gels. The cell wall porin increased the specific conductance of artificial lipid bilayer membranes from phosphatidylcholine/phosphatidylserine mixtures by the formation of cation-selective channels, which had an average single-channel conductance of 3.0 nS in 1 M KCl. The single-channel conductance was only moderately dependent on the bulk aqueous KCl concentration, which indicated negative point charge effects on the channel properties. The analysis of the concentration dependence of the single-channel conductance using the effect of negative charges on channel conductance suggested that the diameter of the cell wall channel is about 1.4 nm. Asymmetric addition of the cell wall porin to lipid bilayer membranes resulted in an asymmetric voltage dependence. The cell wall channel switched into substates, when the cis side of the membrane, the side of the addition of the protein, had negative polarity. Positive potentials at the cis side had no influence on the conductance of the cell wall channel.

Necrotizing fasciitis caused by Vibrio vulnificus: first published infection acquired in Turkey is the second time a strain is isolated in Germany.

Vibrio vulnificus, a marine vibrio, has recently been recognized as a potential human pathogen. It causes human infections with mortality rates up to 60%. Until 1991, most human isolations were reported from the USA, Japan and Taiwan. The second strain isolated in Germany is documented and a significant case of V. vulnificus infection acquired in Turkey is published for the first time.