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J S Brazier

Publications and source records attributed to J S Brazier.

At least 55 records · Page 3Linked to original sources

A rapid test for the presumptive identification of Clostridium perfringens.

A novel rapid method for the identification of colonies of Clostridium perfringens (key iD Lab M Ltd. Bury, UK) was evaluated. The method consists of a test strip containing substrates for pre-formed enzymes selected for optimum differentiation of C. perfringens from other clostridia. One hundred and forty-six strains of clostridia were tested using the key iD strip. The strip successfully confirmed the identity of all 73 strains of C. perfringens tested, and differentiated these from 73 strains of 20 other clostridial species. C. absonum and C. baratii, spedes which are very similar to C. perfringens, could also be differentiated by this method. The key iD strip is recommended for laboratories as a rapid alternative to more conventional tests for presumptive identification of C. perfringens.

Journal Article↗

Identification and antimicrobial resistance patterns of clinical isolates of Clostridium clostridioforme, Clostridium innocuum, and Clostridium ramosum compared with those of clinical isolates of Clostridium perfringens.

Clostridium ramosum, C. innocuum, and C. clostridioforme are frequently isolated from clinical specimens including blood. Because of Gram stain variability, a lack of spores, and atypical colonial morphology, identification of these species is often difficult. Three anaerobe identification kits were evaluated for their abilities to identify these species. For comparison, 11 strains of C. perfringens were evaluated in parallel. By using profile numbers and codebooks, the correct genus and species were identified, as follows: with the RapID ANA II kit, 100% (20 of 20) of C. ramosum isolates, 24% (5 of 21) of C. innocuum isolates, and 50% (10 of 20) of C. clostridioforme isolates; with the AnIDent kit, 60% (12 of 20) of C. ramosum isolates, 28% (6 of 21) of C. innocuum isolates, and 90% (18 of 20) of C. clostridioforme isolates; with the ATB32A kit, 70% (14 of 20) of C. ramosum isolates, 0% (0 of 21) of C. innocuum isolates, and 40% (8 of 20) of C. clostridioforme isolates. Profile numbers that overlapped several species were obtained as follows: with the RapID ANA II kit, 0% of C. ramosum isolates, 76% of C. innocuum isolates, and 40% of C. clostridioforme isolates; with the AnIDent kit 40% of C. ramosum isolates, 62% of C. innocuum isolates, and 5% of C. clostridioforme isolates; with the ATB32A kit, 15% of C. ramosum isolates, 52% of C. innocuum isolates, and 25% of C. clostridioforme isolates. One strain of C. innocuum was misidentified by the AnIDent kit, and the remainder yielded profile numbers that were not listed in the codebooks. The MICs of 11 antimicrobial agents including penicillin G, metronidazole, clindamycin, cefoxitin, cefotetan, imipenem, meropenem, amoxicillin-clavulanate, ampicillin-sulbactam, piperacillin-tazobactam, and vancomycin were determined by the agar dilution method. All C. perfringens strains were susceptible to all antimicrobial agents tested. Various levels of resistance to cefoxitin, cefotetan, and penicillin G were noted with C. ramosum, C. clostridioforme, and C. innocuum. In addition, resistance to clindamycin was noted with C. ramosum (5%) and C. innocuum (10%). Most strains of C. innocuum were only moderately susceptible to vancomycin (MIC at which 90% of strains are inhibited, 4 micrograms/ml).

Bacteriological Techniques↗

Role of the laboratory in investigations of Clostridium difficile diarrhea.

The laboratory diagnosis of acute diarrhea due to Clostridium difficile can be based solely on the demonstration of either cytotoxin or enterotoxin in the stool. This and other noncultural methods, however, fail to provide isolates from which useful data can be obtained. Outbreaks can be recognized only by comparison of isolated strains. A dual approach of toxin detection and isolation is therefore recommended.

Bacterial Proteins↗

Optimal methods for identifying Clostridium difficile infections.

The major controversy in the diagnosis of symptomatic gastrointestinal infection due to Clostridium difficile is whether laboratory evidence of the C. difficile organism in culture is sufficient or if evidence of one of the C. difficile toxins in stool should be required. Cultures performed properly on selective media currently are the most sensitive method for detection of C. difficile, whereas the cell cytotoxin assay for detection of toxin B is the most specific. Stool specimens from patients with clinical diarrhea are sometimes found to be culture-positive for C. difficile but assay-negative for cytotoxin. Samples from these patients can be viewed as false-positive by culture or false-negative by cytotoxin test. Evidence from endoscopy indicates that some patients whose stool is culture-positive for the organism but assay-negative for toxin do have pseudomembranous colitis, but its incidence among such patients (11%) is lower than that among patients whose stool is culture- and assay-positive (51%). Response to treatment with vancomycin or metronidazole is similar in the two groups of patients, and withholding treatment from patients whose stool contains C. difficile but not cytotoxin may result in increased morbidity and mortality. Up to one-third of C. difficile organisms from stool specimens that are culture-positive but assay-negative are incapable of producing cytotoxin in vitro, a finding that suggests these organisms may not be the cause of diarrhea.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins↗

Propionibacterium propionicum and infections of the lacrimal apparatus.

Analysis of the clinical details associated with 26 referred isolates of Propionibacterium propionicum revealed the marked propensity of this anaerobe to be involved with infections of the lacrimal apparatus; 18 (69%) of the 26 referred isolates were from cases of canaliculitis in which this organism was regarded as the significant pathogen. Further analysis of data showed a significant correlation with the sex of the patient. Of the 18 cases of canaliculitis, the sex of the patient was reported in 16; 14 (88%) of these 16 patients were females (P < .005). The age of 13 of the 18 patients with canaliculitis was known (mean, 61.6 years; range, < 1-80 years); it appears from these cases that P. propionicum lacrimal infections are more common in later life. None of the referred isolates were identified as Actinomyces species. Despite the fact that infection of the lacrimal apparatus is known as ocular actinomycosis, it appears that P. propionicum is more frequently associated with the condition than are true Actinomyces species.

Actinomyces↗

An investigation of a nosocomial outbreak of Clostridium difficile by pyrolysis mass spectrometry.

Isolates from a presumptive nosocomial outbreak of Clostridium difficile infection at a large teaching hospital were typed by pyrolysis mass spectrometry (PMS) and antibiograms. One isolate, from the putative index case, was dissimilar from the outbreak strain, but 24 isolates from 16 patients were indistinguishable by both methods. The outbreak centred on two wards for the acute care of the elderly, with a few cases elsewhere. Transfer of patients appeared to be the route of transmission between wards. There was a significant fall in the incidence of cases following intervention by the Infection Control Unit. This included ward inspection, advice on antibiotic usage and advice on prevention of faecal-oral transfer, particularly by proper handwashing. Subsequent monitoring of C. difficile infection showed a background of sporadic, dissimilar isolates with occasional apparent cross-infection incidents limited to a few patients. In suspected outbreaks, patterns of antibiotic susceptibility may be useful in initial screening, before referral for more sophisticated typing. There was excellent correlation between PMS results, antibiograms and epidemiological information.

Aged↗

Evaluation of a commercial enzyme immunoassay kit for the detection of Clostridium difficile toxin A.

A new enzyme immunoassay (EIA) kit developed for the rapid detection of Clostridium difficile toxin A in faecal specimens, Premier (Meridian Diagnostics), was evaluated using 101 faecal specimens. Sixty-nine specimens were positive for Clostridium difficile by isolation of the organism and by cytotoxicity in tissue culture. The EIA for toxin A was positive in 49 of these 69 cases. No specimen that was negative for cytotoxicity was positive by EIA. Eight of the 32 specimens negative by both EIA and cytotoxicity assay yielded Clostridium difficile by culture. In five of these cases the cytotoxigenic status of the isolate was determined, and four were positive. There was no direct relationship between cytotoxin titre and EIA reading.

Animals↗

A selective medium for Fusobacterium spp.

A new selective medium (JVN) for the isolation of Fusobacterium spp. from clinical material is described. The medium incorporates josamycin, vancomycin and norfloxacin (at 3, 4 and 1 microgram/ml, respectively) as the selective agents, plus 5% defibrinated horse blood in Fastidious Anaerobe Agar Base (Lab M). This formula allowed luxuriant growth of all 82 strains (eight recognized species) of fusobacteria tested, while significantly inhibiting 51/51 (100%) strains of facultative anaerobes and 45/51 (88%) strains of other obligate anaerobes. JVN medium allowed the successful isolation of strains of Fusobacterium naviforme, F. nucleatum and F. necrophorum from the gingivae of 9/16 healthy volunteers, and strains of F. varium and F. mortiferum from faecal suspensions seeded with these organisms.

Culture Media↗

Analysis of the porphyrin content of fluorescent pus by absorption spectrophotometry and high performance liquid chromatography.

Extracts of 19 samples of pus which showed red fluorescence with ultraviolet light were screened for the presence of porphyrins by absorption spectrophotometry. All those which showed spectra typical of metal-free porphyrins were analysed by high performance liquid chromatography to identify the porphyrins present. These were predominantly the di-carboxylic porphyrins, deuteroporphyrin and mesoporphyrin, and another which was thought to be pemptoporphyrin. This combination matched those reported previously in normal stools. Protoporphyrin IX was shown not to be the most common fluorescent pigment in pus and was never present alone. However, the di-carboxylic porphyrins may be produced by bacterial metabolism of its labile vinyl side-chains. Black-pigmented bacteroides (the melaninogenicus group of Bacteroides spp. and Porphyromonas spp.) were isolated from 12 (63%) of the 19 pus samples; these may produce protoporphyrin IX by the demetallation of haem.

Bacterial Infections↗

Fastidious anaerobe agar compared with Wilkins-Chalgren agar, brain heart infusion agar, and brucella agar for susceptibility testing of Fusobacterium species.

Fastidious anaerobe agar supported the growth of 82 strains of fusobacteria better than brain heart infusion agar, brucella agar, and Wilkins-Chalgren agar. Fastidious anaerobe agar showed less hazing and fewer tailing endpoints with beta-lactam antibiotics. Whole-blood supplementation improved the performance of all media. Wilkins-Chalgren agar without blood failed to support the growth of 17% of the strains. All Fusobacterium ulcerans strains were resistant to clindamycin.

Agar↗

Evaluation of the Anoxomat: a new technique for anaerobic and microaerophilic clinical bacteriology.

A system of automatic jar evacuation-replacement (Anoxomat) for the culture of anaerobes, capnophiles, and microaerophiles was compared with existing methods of anaerobic cabinets, carbon dioxide incubators, and manual evacuation-replacement. Of the 50 species of anaerobes, 29 strains of capnophiles, and 11 strains of microaerophiles tested, equivalent growth was obtained in all but two instances. The Anoxomat system yielded slightly larger colonies in 26 (52%) of anaerobes tested with superior growth in the anerobic cabinets in three (6%) of cases and equal in both in 21 (42%). Of the microaerophiles and capnophiles tested, there was no significant difference between the Anoxomat and the conventional system. The Anoxomat system seems to be a suitable alternative to anaerobic and carbon dioxide incubators.

Bacteria↗

UV red fluorescence of Veillonella spp.

A total of 34 clinical isolates and 7 type strains of Veillonella spp. were tested for their ability to fluoresce on various culture media. Fluorescence was medium dependent and varied among the species. Scanning absorption spectrophotometry of culture extracts showed that the absorption spectrum of the fluorescent pigment is typical of a metal-free porphyrin.

Culture Media↗

Comparison of alcohol shock enrichment and selective enrichment for the isolation of Clostridium difficile.

Two enrichment methods were compared for their ability to recover Clostridium difficile from stool samples. One method used selective enrichment in an antibiotic-containing broth followed by detection with a latex particle agglutination (LPA) reagent. The other used enrichment in a non-selective broth following treatment of the specimen with alcohol. With clinical specimens enrichment culture was significantly more successful at detecting C. difficile than direct plating. Alcohol shock enrichment was twice as effective as direct culture, while selective broth enrichment was three times more effective. The use of LPA for screening selective enrichment broths for C. difficile should prove a cost-effective measure as only positive broths (about 20%) require subculture for confirmation.

Bacteriological Techniques↗

A note on ultra-violet red fluorescence of anaerobic bacteria in vitro.

Anaerobes other than the Bacteroides melaninogenicus group isolated from clinical material produce an ultra-violet red fluorescence when grown under certain conditions in vitro. These organisms include other members of the genus Bacteroides as well as strains of some species of Clostridium, Bifidobacterium and Actinomyces. The major fluorescent pigment was identified as protoporphyrin IX. Factors necessary for the production of fluorescence are the presence of blood or haem and a fermentable carbohydrate during growth on a solid medium. Fluorescence intensity was related to the concentration of blood and fermentable carbohydrate present but was independent of inoculum size. Certain commercially available blood agar bases designed specifically for the isolation of fastidious anaerobes from clinical material which contain added carbohydrate were shown to induce fluorescence in certain organisms. This may lead to the misidentification of some anaerobes as B. melaninogenicus.

Actinomyces↗