Biomaterial Database

Evaluation of selective media for the isolation of Brachyspira aalborgi from human faeces. 2003

C Josephine Brooke, and Thomas V Riley, and David J Hampson

Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, Western Australia 6150, Australia 2,3Division of Microbiology and Infectious Diseases, The Western Australian Centre of Pathology and Medical Research2 and Department of Microbiology, The University of Western Australia3, Nedlands, Western Australia 6009, Australia#dReceived 5 November 2002 Accepted 2 March 2003.

The purposes of this study were to identify a solid medium that supports improved growth of the anaerobic intestinal spirochaete Brachyspira aalborgi, to modify this for use as a selective isolation medium and then to test the medium for its effectiveness in isolating B. aalborgi from patients' faeces. Of the media evaluated, brain heart infusion agar (BHIA) with 10 % bovine blood (BB) was the most effective base-supplement combination for growth, with colonies attaining 1.2 mm in diameter by 21 days. Incubation in an anaerobic jar (94 % H(2), 6 % CO(2)) permitted growth of larger colonies than incubation in an anaerobic chamber (80 % N(2), 10 % H(2), 10 % CO(2)). Growth was improved only slightly at 38.5 degrees C compared with 37 degrees C. Selection of B. aalborgi from artificially seeded faeces was achieved equally well on eight different solid media containing spectinomycin (400 micro g ml(-1)) alone or in combinations with polymyxin B (5 micro g ml(-1)), colistin (25 micro g ml(-1)) and rifampicin (12.5 micro g ml(-1)). By using BHIA 10 % BB with spectinomycin plus polymyxin B, B. aalborgi was isolated from one of five human faecal samples that were positive for B. aalborgi by PCR amplification. This is the first report of the isolation of B. aalborgi from human faeces.

UI	MeSH Term	Description	Entries
D008854	Microscopy, Electron	Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.	Electron Microscopy
D008858	Microscopy, Phase-Contrast	A form of interference microscopy in which variations of the refracting index in the object are converted into variations of intensity in the image. This is achieved by the action of a phase plate.	Phase-Contrast Microscopy,Microscopies, Phase-Contrast,Microscopy, Phase Contrast,Phase Contrast Microscopy,Phase-Contrast Microscopies
D003470	Culture Media	Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.	Media, Culture
D005243	Feces	Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013144	Spirochaetales	An order of slender, flexuous, helically coiled bacteria, with one or more complete turns in the helix.	Spirochetes,Spirochete
D013145	Spirochaetales Infections	Infections with bacteria of the order SPIROCHAETALES.	Spirochete Infections,Infections, Spirochaetales,Infections, Spirochete,Infection, Spirochaetales,Infection, Spirochete,Spirochaetales Infection,Spirochete Infection
D016133	Polymerase Chain Reaction	In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.	Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain