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.

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.

Actinomycete infections in humans--a review.

Diseases caused by pathogenic aerobic and facultatively anaerobic actinomycetes differ considerably with respect to their etiology, pathogenesis, clinical appearance and epidemiology. Facultatively anaerobic (fermentative) actinomycetes may not only be involved etiologically in the three classical forms of cervicofacial, thoracic and abdominal actinomycoses, but also in infections of the female genital organs, the eye, the tissue adjacent to dental implantation elements and tooth extraction wounds. The species distribution of the fermentative actinomycetes isolated from these conditions varied to a certain, but characteristic, extent, as did the concomitant actinomycotic flora. The sex ratio reported for human Actinomyces infections (male:female = 3:1) appeared to be restricted to actinomycotic abscesses and empyemas. The prevailing pathogenic, obligately aerobic actinomycete species in Germany was found to be Nocardia farcinica followed by Nocardia asteroides. The comparatively high incidence of N. farcinica infections was chiefly due to the occurrence of nosocomial postoperative wound infections by this pathogen observed in two German hospitals. Besides surgical treatment, immunosuppressive treatment appeared to be the most common factor predisposing for nocardiosis. Recent observations strongly suggested that the spectrum of human nocardial infections in Germany has been changing, as regards the overall incidence, the prevalence of N. farcinica, the sex ratio, the mean age of patients, as well as the role of N. farcinica as a possibly important nosocomial pathogen.

Identification of some clinically significant actinomycetes.

Early recognition of infections caused by actinomycetes tend to be highly dependent on at least a tentative diagnosis derived from microbiological tests, since the clinical symptoms can be difficult to interpret. Reliable identification of clinically significant actinomycetes depends upon the application of taxonomic techniques that are not yet widely used in clinical laboratories. The value of rapid enzyme, chemical and molecular fingerprinting techniques is exemplified by their application to the identification of representatives of clinically significant actinomycete taxa.

Medical and microbiological problems arising from airborne infection in hospitals.

The practical importance and frequency of airborne nosocomial infections has been a matter of dispute for many years. This is because most of the pathogens acquired in hospitals are able to use various different routes of infecting the patient's body so that it may be difficult or even impossible to prove an individual infection to be airborne. Only microbes such as Streptococcus pyogenes, Neisseria meningitidis, Corynebacterium diphtheriae, Mycobacterium tuberculosis, or certain respiratory viruses that are known to be predominantly transmitted by droplet infection from infected persons or healthy carriers, have been accepted to the cause of airborne nosocomial infections. Other pathogens such as legionellae, pseudomonads or clostridia may be distributed in the hospital environment via an insufficient or defective air-conditioning system, with or without humidification. The assessment of indirect airborne infections caused by infective particles derived from dust which has settled on furniture or the floor or which has been introduced to the hospital environment by shoes, open windows, building works or potted indoor plants is much more difficult. Many Gram-positive bacteria such as Staphylococcus aureus, mycobacteria, nocardiae, and endospores of clostridia and bacilli, as well as the reproductive elements of fungi do remain viable and infective in dry dust and may therefore infect patients when the dust is disturbed. In contrast to nosocomial infections due to Gram-negative bacteria, against which most preventive measures have been concentrated in the past and which are usually not airborne in origin, it appears that the possibility of direct or indirect transmission of hospital pathogens by air has been underestimated.(ABSTRACT TRUNCATED AT 250 WORDS)

[Preoperative reduction of intestinal bacteria through orthograde lavage of the large intestine]. / Die präoperative Darmkeimverminderung durch orthograde Dickdarmspülung

The orthograde colon lavage, suggested by Crapp, is a new method for the preoperative preparation of the colon. On the day prior to surgery, the sitting patient receives for 4 h through a stomach probe, 101 of physiological saline solution warmed up to 37 degrees C. The result is a complete evacuation of the bowel. If a bactericidal antibiotic with a broad spectrum of activity (neomycin) is added, the suppression of the microflora amounts to 92%. The regeneration of the intestinal flora could be observed after a few days. A complete evacuation of the colon is guaranteed by this procedure, so that all operations can be performed under optimal conditions. From the clinical as well as from the microbiological point of view this method is without any risk. -- The orthograde colon lavage is indicated in every case of colon, rectum and anal surgery, but it is contraindicated in patients with highly stenosing tumors.

[The microbial colonization of implant elements made of plastics and titanium]. / Die mikrobielle Besiedlung von Implantatelementen aus Kunststoff und Titan.

12 edentate subjects wearing lower complete dentures stabilized by an IMZ-implant-supported Dolder bar took part in the experiment. Elastic (POM) and titanium connection parts were alternatingly inserted. After the removal of 24 POM and titanium elements from each of the patients, a qualitative analysis of the microbial flora was performed. Except for actinomycetes, aerobic as well as anaerobic microorganisms were found less frequently with titanium than with POM elements.

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.

[Anaerobic infections in operative medicine]. / Anaerobierinfektionen in der operativen Medizin.

Compared with other medical disciplines, anaerobic infections play an especially important role in operative medicine. In odontogenic soft tissue infections of the orofacial area, in intraabdominal suppurative processes and in post partum, post abortum or postoperative gynecological infections, anaerobic pathogens are almost inevitably encountered. Furthermore, even in times of peace exogenous or endogenous toxic clostridial infections do not occur so rarely that they should be excluded from considerations concerning differential diagnosis. Because of the substantial malignancy of some of these infections, because of considerable variation in the species composition of anaerobic infections at different body sites and because of the peculiar antibiotic sensitivity pattern of certain anaerobes, diseases caused by anaerobic pathogens deserve special diagnostic and therapeutic attention. The reliable recognition of the anaerobic etiology of inflammatory lesions is usually only possible when microscopic or cultural bacteriological examinations are performed. As complete bacteriological analyses may be comparatively time-consuming, the clinician's requirements for planning the appropriate treatment are met best when he is informed of the tentative diagnosis of an anaerobic infection as soon as possible. This may often be achieved by microscopic or gaschromatographic techniques within less than two hours. The treatment of "surgical anaerobic infections" usually consists of operative measures combined with antibacterial chemotherapy. Only in selected cases, incision and drainage will suffice to cure a localized anaerobic suppuration. As a rule, additional application of antibacterial drugs is necessary or at least advisable in order to reliably prevent relapses or systemic spread. Appropriate chemotherapeutics are selected according to the pathogens that are individually present or may be expected with some probability. Diseases due to Gram-positive, non-spore-forming anaerobes, fusobacteria and Bacteroides species of the melaninogenicus group are comparatively easy to treat as these organisms are usually susceptible to penicillins including benzylpenicillin in sufficiently high doses. Against beta-lactamase-producing intestinal Bacteroides species such as B. fragilis and B. thetaiotaomicron, lincomycins and nitroimidazoles have proved to be highly effective. Infections caused by these Bacteroidaceae may also respond to treatment with mezlocillin, cefoxitin, lamoxactam and possibly aminopenicillins combined with clavulanic acid.(ABSTRACT TRUNCATED AT 400 WORDS)

Special methodological problems in antibiotic susceptibility testing of fermentative actinomycetes.

Knowledge on the antibiotic suceptibility of pathogenic fermentative actinomycetes is still fragmentary and contradictory, which is one reason why even today the chemotherapy of human actinomycosis may still present problems. Complete and reliable data on the in vitro sensitivity of these pathogens can be obtained, however, when standardized and specially adapted methods of susceptibility testing are employed. Special adaptation should cover the general technique of in vitro testing, as well as the choice of test media, the preparation of inocula, incubation methods and method of reading results. Provided that all factors are taken into account which might cause difficulties in test reproducibility and therapeutic relevance of the results, in vitro tests appear to contribute to the revision and updating of treatment regimes for actinomycosis. Our findings indicate that, apart from ampicillin the clinical efficacy of which has already often been proved, mezlocillin, cefoxitin, clindamycin, tetracyclines, rifampicin and erythromycin may be recommended as alternative drugs which would seem to offer good chances of success.

A numerical taxonomic study of members of the Actinomycetaceae and related taxa.

Two hundred and twenty-two representatives of the Actinomycetaceae and related taxa, including reference cultures, received strains and fresh isolates, were tested using 124 unit characters. The data were examined using numerical taxonomic techniques with various coefficients and average linkage clustering; the variation in composition of the clusters obtained from the different coefficients was slight. Most species included in the study formed discrete phena, which exhibited good differential characters. Actinomyces israelii was particularly well-defined and A. naeslundii and A. viscosus, although grouping together, did show divergence in several test results. Actinomyces bovis was initially linked with representatives of genera other than Actinomyces, these being Bifidobacterium, Corynebacterium pyogenes and Erysipelothrix rhusiopathiae. The strains of Bacterionema matruchotii and Rothia dentocariosa formed tight distinct clusters associated only at low similarity levels with other members of the family Actinomycetaceae. The Arachnia propionica phenon, although well-defined, appeared to consist of two subclusters which could be assigned to serotypes 1 and 2. Tests which may be useful in identification are tabulated and the relationships between the various taxa are discussed.

[Sporotrichoid infection caused by Nocardia brasiliensis]. / Sporotrichoide Infektion durch Nocardia brasiliensis.

A few days after a spine of one of his indoor cacti had pricked the fourth finger of his left hand, a 56-year-old man developed multiple inflammatory suppurating nodules and infiltrates and a cord-like lymphangitis on the left forearm and upper arm. Nocardia brasiliensis was cultivated from aspirated pus. Subsequently, both Nocardia brasiliensis and Nocardia asteroides were isolated from the cactus earth. This is the first report of an infection by Nocardia brasiliensis acquired in Germany.

Apical growth in facultative Anaerobic actinomycetes as determined by immunofluorescent labeling.

Apical growth, following synthesis of newly formed cell wall material at the poles of rods of Actinomyces israelii, A. naeslundii, Arachnia propionica and Bacterionema matruchotii, has been demonstrated by means of immunofluorescent labeling, confirming previous observations by scanning electron microscopy. Some implications of the phenomenon are discussed.

Rapid micromethods for detecting deamination and decarboxylation of amino acids, indole production, and reduction of nitrate and nitrite by facultatively anaerobic actinomycetes.

96 strains of facultatively anaerobic actinomycetes and 2 Propionibacterium acnes strains were studied for their ability to deaminate and/or decarboxylate 13 amino acids, to reduce nitrate and nitrite, and to produce indole, using specially adapted micro-methods. Several of the tests performed were found to provide information which may aid in improving the classification and in facilitating the identification of these organisms.

Serum antibodies to pathogenic actinomycetes in the normal human population.

Sera of persons without known actinomycotic infection (n = 153) were tested for antibodies reacting with antigenic preparations of different Actinomyces spp. and Nocardia spp. By using an enzyme immunoassay, 16% of all of the sera analyzed reacted significantly with antigens of A. viscosus and 2% those of with A. naeslundii. The antigens detected by these antibodies were of low molecular mass (14-32 kDa) and showed a uniform reaction pattern in the immunoblot analysis. Multiple bands coccurred with a difference of approximately 2 kDa in size suggesting the presence of repetitive units. Analogous antibodies were not observed with A. israelii serovar 1 and serovar 2 antigens, which were only insignificantly bound by the sera. The antibodies demonstrated were most probably acquired during episodes of periodontal disease or gingivitis in which A. viscosus is etiologically involved. Antibodies against nocardial antigens were not demonstrated in significant proportions by enzyme immunoassay and immunoblot analysis.