Turicella otitidis and Corynebacterium auris: 20 years on.

Turicella otitidis and Corynebacterium auris, described as new species 20 years ago, have been isolated mainly from the external ear canal and middle ear fluid. While their taxonomic position has been clearly established, their diagnosis in the routine laboratory is difficult. The question of their pathogenic potential in otitis is still open but might be elucidated better if corynebacteria are speciated more often.

Pseudoepidemic of nontuberculous mycobacteria due to a contaminated bronchoscope cleaning machine. Report of an outbreak and review of the literature.

We describe a pseudoepidemic due to nontuberculous mycobacteria contaminating the water tank of a machine used to clean and disinfect fiberoptic endoscopes. Forty-six bronchoscopies performed on 41 patients during a six-month period yielded 16 specimens positive for acid-fast bacilli (AFB). One specimen showed Mycobacterium avium complex from an AIDS patient and one, M tuberculosis from a patient with active cavitary tuberculosis. In four patients, only the smears showed AFB; subsequent cultures remained negative. Of the rest, seven contained M chelonae and three M gordonae, all in patients with no clinical signs of mycobacterial disease. Two of the three M gordonae isolates represented laboratory contamination from an antimicrobial solution in a culture medium. Four patients in the beginning of the pseudoepidemic were treated for presumed tuberculosis until negative culture results were available. Control of the "outbreak" was achieved by regular disinfection of the implicated water tank in the cleaning machine. Contamination of bronchoscopes with nontuberculous mycobacteria can lead to unnecessary diagnostic and therapeutic interventions.

Grouping of some clinically relevant gram-positive rods by automated fatty acid analysis. Diagnostic implications.

One hundred and ninety-four strains of aerobically growing Gram-positive rods of the genera Corynebacterium, Actinomyces, Arcanobacterium, Erysipelothrix, Listeria, Oerskovia, Nocardia, Rhodococcus, and of unnamed Center for Disease Control (CDC) groups were checked for cellular fatty acid profiles with the microbial Identification System (Microbial ID, Newark, Del., USA). In order to obtain unified data usable for the clinical laboratory, 24 or 48 h sheep blood agar cultures were used. It was thought that grouping and perhaps identification could be aided by this approach. With the aid of numerical analysis, four groups (two consisting of two subgroups each) were established. The discriminatory ability of the scheme, however, was only 77.2%, indicating that grouping of an unknown isolate could be done with some accuracy, but that speciation would still require biochemical testing.

Identification of coryneform and other gram-positive rods with several methods.

The identification of 202 isolates of aerobically growing Gram-positive rods from clinical material was attempted by using a combination of "traditional" morphological and biochemical tests (Hollis & Weaver (20)) plus patterns of cellular and metabolic fatty acids. This system served as the "gold standard" for three others, i.e. API Coryne (Rapid Coryne), MIDI TSBA and MIDI CLIN Aerobic. In addition, several growth, biochemical and susceptibility tests (growth on cystine-tellurite blood agar, DNase, hippurate and starch hydrolysis, methanethiol formation, API ZYM, CAMP reaction, susceptibility to O/129 and to six antimicrobials) were done in order to check their usefulness for the identification of this group of bacteria. Our system, with the help of chemotaxonomic tests (m-DAP and mycolic acids), was able to identify 154/202 (76%) of the isolates by species and an additional 41/202 (21%) by genus only; 7 (3%) could not be identified. The API Coryne system identified to species or genus level 140/195 isolates (72%). Corresponding figures for the MIDI TSBA and CLIN systems were 63/195 (32%) and 88/195 (45%); further details of species and genus identification are presented in the text. The main drawback of the commercial systems is the extent and probably the numerical depth of the data base. We recommend the use of our multisystem approach for the identification of Gram-positive rods until commercial systems are based on a broader and numerically more extensive data base. The additional tests did not prove species- or genus-specific.

Heterogeneity within Corynebacterium minutissimum strains is explained by misidentified Corynebacterium amycolatum strains.

Forty-eight clinical strains that were tentatively identified as Corynebacterium minutissimum on the basis of standard biochemical reactions (Hollis-Weaver tables) as well as by the use of the API (RAPID) Coryne system were examined further. Two different groups of strains were observed. The first group (including the type strain of C minutissimum) contained 27 strains showing creamy colonies. These strains grew homogeneously in 6.5% NaCl broth, exhibited DNase activity, were susceptible to the vibriocidal compound O/129, produced succinic acid, and contained mycolic acids. The second group comprised 21 strains with dry colonies. They grew in clumps at the surface of 6.5% NaCl broth, DNase activity was not detected, they were resistant against O/129, produced large amounts of propionic acid, and mycolic acids were not detected. In combination with quantitative DNA-DNA hybridizations, it was demonstrated that strains of the second cluster belonged, in fact, to C amycolatum. Furthermore, it was observed that a few C minutissimum strains may also ferment mannitol. These data indicate that the clinical microbiologist must be careful not to misidentify C amycolatum strains as C minutissimum.

Branhamella catarrhalis: fatty acid and lipopolysaccharide analysis of an atypical strain from blood culture.

A Branhamella catarrhalis-like organism was isolated from blood cultures; it was atypical in showing negative nitrate and nitrite and positive gamma-glutamylaminopeptidase reactions, in agglutinating with Neisseria meningitidis antisera, and in forming butyrous colonies. Cell-wall fatty-acid and lipopolysaccharide analysis provided evidence that this isolate was an atypical B. catarrhalis.

Stool cultures for Shigella spp: improved specificity by using MacConkey agar with xylose.

A total of 678 stool specimens were cultured on four different agars: on xylose-lysine-desoxycholate agar (XLD), MacConkey agar (Mac), MacConkey agar supplemented with xylose (Mac-X), and Hektoen enteric agar (HE). Isolation rates for shigellae were 77% on HE, 86% on Mac and Mac-X, and 91% on XLD. The specificities of the media were 61% for Mac, 75% for HE, and 78% for XLD and Mac-X. After overnight incubation, Mac-X is much easier to read than XLD, which requires incubation for at least 22 hours. Based on these results and also on the practical aspect that incubation for 22-21 hours does not fit well in our schedule, we now use Mac-X whenever shigellae need to be looked for (i.e. mainly patients with recent travel to tropical countries). As compared to our previous procedure the workload in the laboratory could be reduced by about 20%.

Antimicrobial therapy of infections with aerobic Gram-positive rods.

This review presents data on in vitro susceptibilities of aerobically growing Gram-positive rods and in vivo activities of antibiotics used against Gram-positive rods. While in some instances susceptibility and efficacy are predictable (e.g. penicillin vs. Listeria and microaerophilic coryneforms, or metronidazole vs. Gardnerella) susceptibility testing by dilution techniques seems necessary for many Gram-positive rods as long as they are deemed clinically relevant.

Rothia dentocariosa: taxonomy and differential diagnosis.

As recent external quality control results have shown, the diagnosis of Rothia dentocariosa infection still presents problems for clinical laboratories. This review describes the taxonomy, as well as the chemotaxonomic, morphological and biochemical characteristics, of this organism, and surveys bacteria that may be confused with Rothia dentocariosa.

Natural antibiotic susceptibility of Listeria species: L. grayi, L. innocua, L. ivanovii, L. monocytogenes, L. seeligeri and L. welshimeri strains.

OBJECTIVE: To investigate the natural susceptibility to 71 antimicrobial agents of 103 Listeria strains belonging to all known Listeria species (L. monocytogenes (N = 21), L. innocua (N = 21), L. seeligeri (N = 21), L. ivanovii (N = 19), L. welshimeri (N = 11), and L. grayi (N = 10)). METHODS: MICs were determined using a microdilution procedure in H-Medium. RESULTS: All listeriae were naturally sensitive or intermediate to tetracyclines, aminoglycosides, penicillins (except oxacillin), loracarbef, cefazoline, cefaclor, cefotiam, cefoperazone, carbapenems, macrolides, lincosamides, glycopeptides, dalfopristin/quinupristin, chloramphenicol and rifampicin (probably except L. grayi). Listeria spp. were naturally resistant or intermediate to most 'modern' cephalosporins (cefetamet, cefixime, ceftibuten, ceftazidime, cefdinir, cefpodoxime, cefotaxime, ceftriaxone, cefuroxime), aztreonam, pipemidic acid, dalfopristin quinupristin and sulfamethoxazole. Significant differences in natural susceptibility among the species were seen with the quinolones, trimethoprim, co-trimoxazole, rifampicin, fosfomycin and fusidic acid. It seems likely that L. grayi is naturally resistant to all antifolates; the species was least susceptible to rifampicin and most susceptible to quinolones, whereas L. ivanovii was naturally resistant to most quinolones. L. ivanovii was naturally sensitive to fosfomycin, whereas L. innocua and L. monocytogenes were naturally resistant. L. ivanovii was also the most susceptible species to fusidic acid. CONCLUSIONS: The present study describes a database on the natural susceptibility of Listeria spp. to a wide range of antibiotics, which can be used to validate susceptibility testing results of these microorganisms.

Can the API (RAPID) Coryne system be used for identification of rapidly growing mycobacteria?

Fifty-six strains of rapidly growing mycobacteria (RGM) and 14 strains of aerobic actinomycetes as quality controls (QC) were tested in the API (RAPID) Coryne system version 2. Both groups yielded codes with low identification scores, considerable overlaps, and similar diagnoses. No species-specific codes were observed. Thus, the system would not be useful for the identification of RGM.

Fulminant postsurgical Bacillus cereus meningitis: case report.

A 25-year-old woman presented with decreased level of consciousness, bilateral papilledema, and bitemporal hemianopsia. While receiving oxacillin prophylaxis, she underwent ventriculostomy and a transsphenoidal approach for the removal of a growth hormone- and prolactin-secreting adenoma of the pituitary. Within 4 days, fewer, symptoms of meningitis, and marked cerebrospinal fluid (CSF) pleocytosis developed, associated with many large Gram-positive rods in the CSF, subsequently identified as Bacillus cereus. This case emphasizes the potential for Bacillus species to cause serious disease following surgery, including meningitis after intracranial surgery. Meningitis may be severe, and organisms are often resistant to standard surgical prophylactic regimens, which might include penicillin or cephalosporin derivatives. Isolation of Bacillus species from the CSF requires evaluation; these organisms should not be dismissed as contaminants or "non-pathogens," particularly when isolated from CSF of patients who have recently undergone neurosurgical procedures.

Differential diagnosis between Streptococcus agalactiae and Listeria Monocytogenes in the clinical laboratory.

Streptococci of the group B (S. agalactiae) and Listeria monocytogenes resemble each other in many morphological and biochemical characteristics. Ten beta-hemolytic strains of each species were subjected to 26 tests commonly and easily performed in the clinical laboratory. Macroscopic and microscopic morphology on solid media showed differences only in the size of the colonies and in the length of the individual organisms. Among many other tests, hippurate hydrolysis and the CAMP reaction were positive in both species. In the presence of these two reaction, a negative catalase test and chaining in broth would make a presumptive diagnosis of S. agalactiae, while motility at 25 C, the presence of the Henry effect, and resistance to furadantin would be indicative of L. monocytogenes.