A plasmalogen, plasmenylethanolamine, is required for in vitro growth of strains of Eubacterium which convert cholesterol to coprostanol. Plasmenylethanolamine was isolated from calf brain by selective saponification of lipid fractions separated by thin-layer or column chromatography. Cholesterol-containing thioglycolate broth plus purified plasmenylethanolamine or its 2-lyso derivative supported growth of Eubacterium ATCC 21408 and a cholesterolreducing Eubacterium isolated from baboon feces. Plasmenylethanolamine obtained from commercial sources also supported growth of these organisms, but none of a number of other pure lipids would support growth. Metabolism of the alkenyl ether group of plasmenylethanolamine occurred during growth.

We characterized two isolates of cholesterol-reducing Eubacterium by conducting conventional biochemical tests and by testing various sterols and glycerolipids as potential growth factors. In media containing cholesterol and plasmenylethanolamine, the tests for nitrate reduction, indole production, and gelatin and starch hydrolyses were negative, and no acid was produced from any of 22 carbohydrates. Both isolates hydrolyzed esculin to esculetin, indicating ,B-glycosidase activity. In addition to plasmenylethanolamine, five other lipids which contain an alkenyl ether residue supported growth of Eubacterium strain 403 in a lecithin-cholesterol base medium. Of six steroids tested, cholesterol, cholest-4en-3-one, cholest-4-en-3,8-ol (allocholesterol), and androst-5-en-3,B-ol-17-one supported growth of Eubacterium strain 403. All four steroids were reduced to the cultures in base medium without carbohydrate. Nitrate reduction, indole production, and starch, gelatin, and esculin hydrolyses were tested as indicated by Holdeman and Moore (8).

The AutoMicrobic system (AMS) Enterobacteriaceae-plus Biochemical Card was developed to identify a select group of 10 species of glucose-nonfermentative and oxidase-positive fermentative gram-negative bacilli. In this study, 159 nonenteric clinical isolates were identified by the AMS and conventional tube biochemicals based on E. 0. King's (Centers for Disease Control) identification schema. The AMS properly identified 96.7% (117 of 121) of isolates whose taxa were included in the AMS data base. Of 38 isolates (94.7%) in which taxa were not included in the data base, 36 were correctly called unidentified organisms. A principal advantage of the AMS is the automated identification of frequently isolated nonenterics in a period of only 8 to 13 h. The AMS, with the use of the Enterobacteriaceae-plus Biochemical Card appears to be a rapid and accurate system for the identification of the most commonly isolated nonfermentative and oxidase-positive fermentative gram-negative bacilli.

We isolated and characterized nine new strains of cholesterol-reducing bacteria from feces and intestinal contents of baboons. Cholesterol-brain agar was used for the primary isolation, and subsequent biochemical tests were done in a lecithincholesterol broth containing plasmenylethanolamine and various substrates. All strains had similar colony and cell morphology, hydrolyzed the ,-glucosides esculin and amygdalin, metabolized pyruvate, and produced acetate and acetoin. Unlike previously reported strains, the nine new strains did not require cholesterol and an alkenyl ether lipid (e.g., plasmalogen) for growth; however, only two strains reduced cholesterol in the absence of the plasmalogen. These two strains also produced succinate as an end product. Carbohydrate fermentation was variable; some strains produced weak acid (pH 5.5 to 6.0) from only a few carbohydrates, whereas other strains produced strong acid reactions (pH c 5.5) from a wide variety of carbohydrates.

Vibrios were isolated in pure culture from the hemolymph of 7 out of 28 dead or dying aquarium lobsters which had been acclimated to 20-22 degrees C. One isolate was identified as Vibrio parahaemolyticus, one as a related marine Vibrio (probably V. marinus), and five as Vibrio alginolyticus. No isolates of halophilic Vibrio species were made from healthy lobsters using thiosulfate citrate bile salts sucrose agar (TCBS).