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A W Brinkley

Publications and source records attributed to A W Brinkley.

9 recordsLinked to original sources

Isolation and characterization of new strains of cholesterol-reducing bacteria from baboons.

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 lecithin-cholesterol broth containing plasmenylethanolamine and various substrates. All strains had similar colony and cell morphology, hydrolyzed the beta-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 less than or equal to 5.5) from a wide variety of carbohydrates.

Animals↗

Comparison of the AutoMicrobic system and a conventional tube system for identification of nonfermentative and oxidase-positive gram-negative bacilli.

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. O. 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.

Bacteriological Techniques↗

Biochemical characterization of cholesterol-reducing Eubacterium.

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 beta-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-4-en-3-one, cholest-4-en-3 beta-ol (allocholesterol), and androst-5-en-3 beta-ol-17-one supported growth of Eubacterium strain 403. All four steroids were reduced to the 3 beta-ol, 5 beta-H products. The delta 5 steroids cholest-5-en-3 alpha-ol (epicholesterol) and 22,23-bisnor-5-cholenic acid-3-beta-ol were not reduced and did not support growth of the Eubacterium strain.

Carbohydrate Metabolism↗

Plasmenylethanolamine: growth factor for cholesterol-reducing Eubacterium.

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 cholesterol-reducing 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.

Cholesterol↗

Anaerobic fecal bacteria of the baboon.

The predominant bacterial genera of baboon feces were enumerated and identified by established procedures. The predominant genera isolated were Lactobacillus, Eubacterium, Streptococcus, and Bacteroides.

Anaerobiosis↗

Method for evaluating broth culture media: application to Haemophilus.

A method was devised to test the growth-promoting ability of a broth medium. The "dilute to extinction" method determines the inoculum required to develop heavy turbidity in a broth with overnight incubation. A statistical method using Poisson distribution was used to show that a single Haemophilus cell can develop heavy turbidity in an optimal broth. The dilute to extinction method was used to evaluate the shelf life of stored media, to titrate the growth factor requirements of Haemophilus, and to evaluate the use of purified hemin and nicotinamide adenine dinucleotide in a broth medium for the growth of Haemophilus. Of the media tested, the most suitable formulation was Mueller-Hinton broth supplemented with 10 microgram of hemin and 10 microgram of nicotinamide adenine dinucleotide per ml. The dilute to extinction method appears to be especially useful in the development of broth media for fastidious organisms. The method could also be used to assure the quality of other broth media which are required to support the growth of small inocula in the clinical or research laboratory.

Culture Media↗

Growth of cell wall-defective variants of Escherichia coli: comparison of aerobic and anaerobic induction frequencies.

A method for quantitating the conversion of Escherichia coli to colony-forming, cell wall-defective (CWD) bacteria has been developed. The induction frequency, i.e., the percentage of the population recovered as CWD colonies was determined for 20 randomly selected clinical isolates of E. coli under aerobic and anaerobic incubation conditions. Penicillin (1,000 U/ML) was the inducing agent. The 20 strains segregated into three groups. Group I organisms produced CWD colonies with high frequency both aerobically and anaerobically. Grout II organisms showed a much higher induction frequency anaerobically than aerobically. Group III organisms were poor inducers. Thirty percent of the strains were group I, 50% were group II, and 20% were group III organisms. These data indicate that anaerobic conditions enhance the induction and growth of CWD E. coli in the research laboratory and suggest that anaerobic incubation may be important in recovery of medically significant CWD bacteria.

Aerobiosis↗

The isolation of Vibrio parahaemolyticus and related vibrios from moribund aquarium lobsters.

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).

Animals↗