Meningitis caused by Pseudomonas maltophilia.

A case of meningitis caused by Pseudomonas maltophilia is described, which was unusual in that it appeared to lack the predisposing factors commonly associated with this organism. Attention is drawn to the difficulties which may be encountered in the identification of Ps. maltophilia.

Typing of Streptococcus pyogenes by pyrolysis mass spectrometry.

Strains of Streptococcus pyogenes from an outbreak in an oncology ward (13) and routine isolates from sporadic cases (6) were examined blind by pyrolysis mass spectrometry (Py-MS), extending previous work on epidemiological typing. This outbreak appeared more complex than one reported previously, but Py-MS and conventional typing results were in complete agreement. The results confirm the potential of Py-MS as a rapid method for identification at strain level in studies of cross infection.

Classification of oral pigmented anaerobic bacilli by pyrolysis mass spectrometry and biochemical tests.

Clinical (66) and reference (5) strains of pigmented gram-negative anaerobic bacilli, identified as Prevotella intermedia (47), Pr. melaninogenica (1), Pr. corpora (8), Porphyromonas asaccharolyticus (12), P. endodontalis (1) and P. gingivalis (2), were examined by pyrolysis mass spectrometry (PMS) and in conventional tests. Numerical classification based on conventional test reaction patterns (CTRPs) resolved five clusters, four comprising strains identified as Pr. intermedia, Pr. corpora, Pr. melaninogenica, and P. gingivalis respectively, and one comprising strains identified as P. asaccharolyticus and P. endodontalis. Numerical classification based on PMS showed a similar division, with decreasing homogeneity of chemical composition in the order Pr. intermedia, Pr. corpora, P. asaccharolyticus, which agreed with the order of homogeneity in CTRPs. PMS clusters corresponding to the genus Porphyromonas were clearly distinct from those of the genus Prevotella. PMS and CTRP classification disagreed on cluster membership for six strains. PMS identification from blind challenge sets was in agreement with conventional identification for 64 of 67 strains.

Epidemiological typing of Streptococcus pyogenes by pyrolysis mass spectrometry.

Strains of Streptococcus pyogenes from an outbreak of infection on a burns unit (15), a collection of routine isolates from another hospital(12) and isolates from a national survey of throat infections in children in the community(4) were examined blind by pyrolysis-mass spectrometry (Py-MS). The outbreak strains (M22 T12) previously found to give identical typing results in conventional tests, formed a closely similar cluster and were distinct from other hospital and community strains. One hospital and one community strain were loosely associated with this cluster. Another cluster comprised six antibiotic-susceptible strains and two community strains. Six strains did not fall within the clusters; four were antibiotic-resistant strains isolated in hospital, one an antibiotic-resistant strain isolated in the community, and one a susceptible hospital strain. Results show that Py-MS is a potentially valuable method for rapid comparison of strains in studies of infection.

Identification of staphylococci by pyrolysis gas-liquid chromatography.

The identification of bacteria by pyrolysis gas-liquid chromatography (PGLC) was evaluated with the genus Staphylococcus. A matrix containing the characteristics of seven species of Staphylococcus was constructed from the results obtained with 42 named strains identified by biochemical methods. An identification program based upon this database was evaluated in tests with 100 isolates of catalase-positive oxidase-negative gram-positive cocci identified by biochemical and PGLC systems. Agreement between the two systems was 90%. Pure cultures of unknown isolates were identified within 30 min by PGLC using a microcomputer data acquisition and analysis program. A taxonomic analysis of the data is presented, and methods for quality control of PGLC, prolongation of column life, and data analysis are described and discussed.

A pyrolysis mass spectrometry study of the non-pigmented Prevotella species.

Collection strains (21) and non-pigmented clinical isolates (96) provisionally identified as Prevotella spp. were classified numerically on the basis of pyrolysis mass spectrometry (PMS) data and reaction patterns in conventional tests (CTRPs) for volatile and non-volatile fatty acids, pre-formed enzymes and biochemical activity. PMS and CTRP classifications were compared with a previous classification based on visual analysis of SDS-PAGE patterns. Although the order of clusters differed, cross-tabulation of cluster membership revealed strong correlations between classifications. Cluster membership in the PMS classification correlated particularly well with SDS-PAGE results. CTRP clusters corresponded largely to the recognised species of Prevotella, but PMS and SDS-PAGE divided two species into sub-groups: two in P. buccae and five in P. veroralis. The latter subgroups could be discriminated by small but consistent differences in CTRPs. An undesignated, well differentiated cluster of strains appeared closest to the main group of P. buccae strains in PMS and CTRPs. B. (P.) capillus could not be distinguished from P. buccae; these species are regarded as synonymous. Strains of P. zoogleoformans and B. (P.) pentosaceus were well separated from other strains in PMS. A complex comprising clusters of P. disiens, P. oralis, P. veroralis, P. loescheii and a further undesignated group similar to P. melaninogenica was well differentiated from P. buccae and P. oris in PMS; clusters corresponding to P. bivia, P. corporis, P. intermedia and P. denticola formed another complex.

A pyrolysis-mass spectrometry study of Corynebacterium spp.

Clinical (66) and collection (38) strains of Corynebacterium spp., including C. jeikeium and CDC group D2, and of Listeria monocytogenes were examined. Conventional characters used in species identification were assessed by a microbiochemical method, and pyrolysis-mass spectrometry (Py-MS) was performed with a Horizon Instruments PYMS 200X. Classification based on Py-MS data yielded clusters that corresponded with species identification and classification groups from conventional data. One small group of clinical strains, homogeneous in conventional tests and Py-MS, comprised isolates from sputum samples from patients undergoing ventilation; they were similar to collection strains of C. renale and C. striatum; the latter species has been implicated in chest infection. Another group, similar to C. minutissimum in both systems, comprised clinical strains isolated from urogenital specimens. L. monocytogenes strains were clearly distinct from Corynebacterium spp. Groups comprising CDC D2 strains and C. jeikeium were resolved, and were similar to other Corynebacterium spp. Two collection strains of C. xerosis were distinct in conventional tests and Py-MS.

A comparison of Bacteroides ureolyticus isolates from different clinical sources.

Clinical isolates of corroding, gram-negative, anaerobic bacilli (provisionally identified as Bacteroides ureolyticus) from superficial ulcers and soft tissue infections (15), non-gonococcal, non-chlamydial urethritis (12) and adult periodontal disease (14) were compared with reference strains of B. ureolyticus, B. gracilis and Wolinella recta in a series of conventional tests of morphology, biochemical activity, tolerance of dyes and bile salts, and antibiotic sensitivity, gas-liquid chromatographic analysis of metabolic products, and in whole-cell analysis by pyrolysis-mass spectrometry (Py-MS). A numerical taxonomic approach was used with the results of conventional tests and the grouping obtained was compared with that obtained by Py-MS. All the ulcer and soft-tissue isolates and the urethritis isolates were oxidase- and urease-positive and formed a homogeneous set consistent with the reference strain of B. ureolyticus. The dental isolates differed from B. ureolyticus strains and were heterogeneous amongst themselves. None corresponded with the reference strains of B. gracilis or W. recta. The conventional and Py-MS approaches to characterisation produced similar groupings and each distinguished between a single cluster of ulcer-urethritis strains and several clusters of dental strains, although the dendrograms derived from the two approaches differed in the order of the clusters; in the Py-MS dendrogram one subcluster of four dental strains came within the main ulcer-urethritis cluster and a cluster of five ulcer strains was separated as a distinct group.

A pyrolysis mass spectrometry study of fusobacteria.

Strains of fusobacteria (143) were examined by pyrolysis mass spectrometry (Py-MS) with a Horizon Instruments PYMS 200X. Fusobacterium necrogenes, F. necrophorum, F. nucleatum, F. mortiferum, F. varium, F. gonidiaformans, F. naviforme, F. russii and Leptotrichia buccalis were discriminated. Strains of fusobacteria isolated from tropical ulcers, although similar to F. mortiferum in conventional tests, were discriminated from each of these species in Py-MS. Identification of 416 spectra to species level agreed with conventional bacteriological methods in 91.8% of cases, was equivocal in 3.4% and disagreed in 4.8%. Classification based on pyrolysis data resolved groups largely corresponding to the recognised species. However, F. nucleatum strains were divided between two distinct groups. The tropical ulcer strains were resolved as a distinct homogeneous group. Py-MS is a rapid, inexpensive and convenient procedure for characterisation of bacteria, with the capacity for a high throughput of samples, although the initial cost of the apparatus is high.

A numerical taxonomic study of the "Streptococcus milleri" group based upon conventional phenotypic tests and pyrolysis mass spectrometry.

Clinical strains presumptively identified as Streptococcus milleri (60), and blind coded collection strains (21) were characterised in conventional tests and pyrolysis mass spectrometry. Comparison of the clusters found by these two approaches revealed five clearly distinct centres of variation. Three corresponded to the DNA homology groups suggested by Whiley and Hardie (1989) as representing the species S. anginosus, S. intermedius and S. constellatus; a fourth comprised three Lancefield group C beta-haemolytic strains; the fifth may represent a biotype of S. anginosus. The characteristics of the latter group are described.

Classification of black-pigmented anaerobes by pyrolysis mass spectrometry (PMS) and conventional tests (CTs).

Clinical (66: dental 53; vaginal 4; wound 9) and reference (5*) strains of pigmented Gram-negative anaerobic bacilli were examined in pyrolysis mass spectrometry (PMS) and conventional tests (CTs). The strains were identified in CTs as: Prevotella intermedia (48*); Pr. melaninogenica (1); Pr. corporis (7); Porphyromonas asaccharolytica (12*); P. endodontalis (1*) and P. gingivalis (2*). Numerical classification based on CTs resolved five clusters comprising strains identified as (I) Pr. corporis, (II) Pr. melaninogenica, (III) Pr. intermedia, (IV) P. gingivalis and (V) P. asaccharolytica and P. endodontalis. Numerical classification based on PMS showed a similar division, with decreasing homogeneity in the order Pr. intermedia, Pr. corporis, P. asaccharolytica, in agreement with the ordering of homogeneity for these species in CTs. PMS clusters corresponding the Porphyromonas spp. were clearly distinct from those of Prevotella spp. PMS and CT classifications disagreed on cluster membership for only six of the strains. PMS identification from blind challenge sets agreed with conventional identification for 64 of 67 strains.

Micrococcus and Stomatococcus spp. from human infections.

Infections with Micrococcus spp. in six patients and Stomatococcus mucilaginosus in one patient are described. Two of the Micrococcus infections occurred in leukaemic patients with indwelling lines, six episodes occurred in three patients undergoing continuous ambulatory peritoneal dialysis and one occurred in a patient with a ventriculo-peritoneal shunt. Stomatococcus was isolated from fluid draining from a sub-dural haematoma. Colony morphology, oxidase reaction and resistance to nitrofuration were useful in differentiation of micrococci from staphylococci. Incidence and risk factors for micrococcal infections appear similar to those for infections with coagulase-negative staphylococci (CNS), but the literature is confused due to changes in the definitions of these genera.

A numerical taxonomic study of Streptococcus sanguis, S. mitis and similar organisms using conventional tests and pyrolysis mass spectrometry.

Numerical taxonomic methods were applied to the results of pyrolysis mass spectrometry (Py-MS) and conventional test reaction patterns (CTRPs) obtained with 46 strains of oral streptococci identified as representative of six species. Of these: Streptococcus gordonii strains formed a single homogeneous well-differentiated cluster both in the Py-MS and CTRP analyses; S. oralis strains were more diverse, forming 2 CTRP and 3 Py-MS clusters; S. crista, S. parasanguis and S. sanguis formed clear, but diverse and poorly differentiated clusters on the basis of CTRPs, and were poorly differentiated in the Py-MS analysis; and S. mitis strains were clearly diverse in both analyses. Peritrichously fimbriate ('tufted') S. mitis strains formed a tight cluster, well differentiated from the type strain in both Py-MS and CTRP analyses, and may represent a previously unrecognised centre of variation.

A pyrolysis mass spectrometry study of temperature-dependent compositional shifts in Xanthomonas (Pseudomonas) maltophilia.

Xanthomonas (Pseudomonas) maltophilia strains are frequently susceptible to aminoglycoside antibiotic and polymixin B when incubated at 37 degrees C, but resistant at 30 degrees C. Five strains showing temperature dependent resistance, and five that did not were examined by pyrolysis-mass spectrometry, a characterisation method that gives fingerprint data reflecting cell composition. Cultures grown at 30 degrees C and 37 degrees C in the absence of antibiotic were analysed. Strains showing temperature-dependent resistance exhibited a characteristic compositional difference between cells grown at the two temperatures, whereas strains not showing this type of resistance varied widely in the extent and nature of temperature-dependent differences in composition. The precise nature of the chemical differences cannot be elucidated at present, but could be determined by examining purified cell constituents.

Applications of pyrolysis mass spectrometry in studies on the mode of action of antimicrobial agents.

Changes in overall cell composition during exposure to antimicrobial agents were investigated by pyrolysis mass spectrometry (Py-MS) in parental (O+K+), K antigen deficient (O+K-), and O and K antigen deficient (O-K-) strains of Klebsiella aerogenes NCTC 5055. Changes followed distinct patterns that correlated with mode of action: penicillin binding protein (PBP) 3 blockade (ceftazidime, piperacillin); PBP 2 blockade (imipenem); membrane disruption (colistin); bacteriostatic (chloramphenicol), and bactericidal (gentamicin) protein synthesis inhibition; and DNA synthesis inhibition (ciprofloxacin). Changes were pronounced after exposure for 1 hour at 10 x MIC. In general, the O-K- strain showed the largest compositional shifts, and the parental O+K+ strain the least shifts, correlating with bactericidal kinetic studies. Py-MS may be useful in studies of the mode of action of antimicrobial agents, particularly in early screening of new compounds, and in rapid detection of the effects of antimicrobial agents on micro-organisms.

Pyrolysis mass spectrometry as a method for inter-strain discrimination of Candida albicans.

A claim that Candida albicans strains NCPF 3153 and B311 were identical was investigated. Authentic strains were shown to be distinct (P less than 0.1%) by pyrolysis mass spectrometry (PyMS). Of twelve strains, provided as clones of NCPF 3153, seven were authenticated, one yielded an equivocal result and four were distinct from both NCPF 3153 and B311. Of eight B311 clones, six were authenticated and two yielded equivocal results. Although five non-C. albicans yeast strains were identified as distinct from B311 and NCPF 3153, Torulopsis glabrata NCPF 3240 was identified as B311, and one clinical isolate of C. albicans as NCPF 3153. This could be explained by the specificity of the mathematical analysis for discrimination between the authentic strains.