THE isolation rate of anaerobic bacteria from clinical specimens increased manyfold in our laboratory when the techniques for anaerobic bacteriology were revised. We present here an account of our findings with these methods applied as a routine to clinical specimens, and make recommendations concerning the techniques of incubation, media and the use of certain identification tests. The isolation of anaerobic species from different anatomical sites is described, and special attention is given to the yield of organisms from empyema fluid, the peritoneal cavity, surface lesions and blood. MATERIALS AND METHODSWith the general exception of urines, faeces, respiratory specimens and cerebrospinal fluid (CSF), specimens of exudates received between 1 Dec. 1974 and 31 Jan. 1976 were cultured for anaerobic bacteria as described below. Transport of specimens. Samples of pus were transported to the laboratory in sterileMcCartney bottles. Swabs of purulent material were sometimes transported deep in a tube of Cary and Blair transport medium (Cary and Blair, 1964), but many swabs were received in the ordinary dry state.Macroscopic examination. Specimens were examined for colour, odour, consistency and the presence of sulphur granules. Liquid pus samples were examined for brick-red fluorescence under an ultraviolet light (Blak-Ray UVL 21, Ultraviolet Products Inc., San Gabriel, California) at a distance of 6-9 in. (15-23 cm) for presumptive evidence of the presence of Bacteriodes melaninogenicus.Microscopic examination. Samples of pus were examined by gram stain to ascertain the number and proportion of morphological forms present.Culture of clinical material. Our complete range of media used for culture in this study were as follows :1. Blood agar: Columbia Agar Base (London Analytical and Bacteriological Media) with 5 % defibrinated horse-blood; this was obtained as poured plates from London Analytical and Bacteriological Media Ltd, Salford, Lancs. (LAB M). 2. Supplemented brucella agar: Brucella Agar (Difco Laboratories Ltd, West Molesey, Surrey) with 5 % defibrinated horse blood and menadione 0.5 pg per ml (Sigma Chemical Co. Ltd, Kingston-upon-Thames, Surrey). 3. Supplemented selective brucella agar: supplemented brucella agar rendered selective by adding kanamycin 75 pg per ml (Kantrex solution, Bristol Laboratories, Langley, Bucks).
SUMMARY Paper discs containing 5 /tg of novobiocin were used as a presumptive test to differenitiate peptococci and peptostreptococci. Zone diameters were measured and minimum inhibitory concentrations (MIC) of the antibiotic for each group were performed to ascertain the activity of the antibiotic against these genera. All strains of peptococci showed no zone of inhibition in the disc test together with an MIC of 25 ,ug/ml or greater. All strains of peptostreptococci showed zones of inhibition of at least 15 mm diameter together with an MIC of 1 f6 jug/ml or less.
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