Gelatin agar medium for detecting gelatinase production by anaerobic bacteria. 1982

D N Whaley, and V R Dowell, and L M Wanderlinder, and G L Lombard

A new medium, Lombard-Dowell gelatin agar, was developed for detecting gelatinase activity by anaerobic bacteria. The medium contained: Trypticase (BBL Microbiology Systems), 5.0 g; yeast extract (Difco Laboratories), 5 g; sodium chloride, 2.5 g; sodium sulfite, 0.1 g; L-tryptophan, 0.2 g; L-cystine, 0.4 g; hemin, 10.0 mg; vitamin K1, 10.0 mg; agar, 20.0 g; D-glucose, 1.0 g; gelatin, 4.0 g; and distilled water to 1 liter. The pH was adjusted to 7.5. The medium was dispensed in 100- by 15-mm quadrant plastic dishes (5 ml per quadrant). To test for gelatinase activity, we inoculated the medium with a young enriched thioglycolate or chopped meat glucose broth culture or a turbid cell suspension in Lombard-Dowell broth, using a sterile cotton swab, and incubated it under anaerobic conditions for 48 h at 35 degrees C. The quadrants were then flooded with Frazier solution, and clear zones around the bacterial growth were recorded as positive for gelatinase activity. The new medium was tested with a variety of anaerobic bacteria, and the results were compared with data obtained with the conventional technique for detecting gelatinase activity. Overall, there was satisfactory agreement between the two tests in the detection of gelatinase activity, but the Lombard-Dowell gelatin agar tests was more rapid and somewhat more sensitive than the conventional test.

UI MeSH Term Description Entries
D010434 Pepsin A Formed from pig pepsinogen by cleavage of one peptide bond. The enzyme is a single polypeptide chain and is inhibited by methyl 2-diaazoacetamidohexanoate. It cleaves peptides preferentially at the carbonyl linkages of phenylalanine or leucine and acts as the principal digestive enzyme of gastric juice. Pepsin,Pepsin 1,Pepsin 3
D010460 Peptococcus A genus of gram-positive, anaerobic, coccoid bacteria that is part of the normal flora of the mouth, upper respiratory tract, and large intestine in humans. Its organisms cause infections of soft tissues and bacteremias.
D011424 Propionibacterium A genus of gram-positive, rod-shaped bacteria whose cells occur singly, in pairs or short chains, in V or Y configurations, or in clumps resembling letters of the Chinese alphabet. Its organisms are found in cheese and dairy products as well as on human skin and can occasionally cause soft tissue infections.
D003013 Clostridium A genus of motile or nonmotile gram-positive bacteria of the family Clostridiaceae. Many species have been identified with some being pathogenic. They occur in water, soil, and in the intestinal tract of humans and lower animals.
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
D005780 Gelatin A product formed from skin, white connective tissue, or bone COLLAGEN. It is used as a protein food adjuvant, plasma substitute, hemostatic, suspending agent in pharmaceutical preparations, and in the manufacturing of capsules and suppositories. Gelafusal
D000362 Agar A complex sulfated polymer of galactose units, extracted from Gelidium cartilagineum, Gracilaria confervoides, and related red algae. It is used as a gel in the preparation of solid culture media for microorganisms, as a bulk laxative, in making emulsions, and as a supporting medium for immunodiffusion and immunoelectrophoresis.
D000693 Anaerobiosis The complete absence, or (loosely) the paucity, of gaseous or dissolved elemental oxygen in a given place or environment. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed) Anaerobic Metabolism,Anaerobic Metabolisms,Anaerobioses,Metabolism, Anaerobic,Metabolisms, Anaerobic
D001419 Bacteria One of the three domains of life (the others being Eukarya and ARCHAEA), also called Eubacteria. They are unicellular prokaryotic microorganisms which generally possess rigid cell walls, multiply by cell division, and exhibit three principal forms: round or coccal, rodlike or bacillary, and spiral or spirochetal. Bacteria can be classified by their response to OXYGEN: aerobic, anaerobic, or facultatively anaerobic; by the mode by which they obtain their energy: chemotrophy (via chemical reaction) or PHOTOTROPHY (via light reaction); for chemotrophs by their source of chemical energy: CHEMOLITHOTROPHY (from inorganic compounds) or chemoorganotrophy (from organic compounds); and by their source for CARBON; NITROGEN; etc.; HETEROTROPHY (from organic sources) or AUTOTROPHY (from CARBON DIOXIDE). They can also be classified by whether or not they stain (based on the structure of their CELL WALLS) with CRYSTAL VIOLET dye: gram-negative or gram-positive. Eubacteria
D018093 Gelatinases A class of enzymes that catalyzes the degradation of gelatin by acting on the peptide bonds. EC 3.4.24.-. Gelatinase

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