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Biomedical subjects

D S Kellogg

Publications and source records attributed to D S Kellogg.

At least 19 recordsLinked to original sources

Cultivation of Neisseria gonorrhoeae under low-oxygen conditions.

Sixteen Neisseria gonorrhoeae strains were examined for their abilities to grow under reduced oxygen tension. Low oxygen tensions were developed by evacuation-replacement procedures in which anaerobes and oxidation-reduction indicators were used as controls. All strains survived 96 h in a medium reduced to below -125 mV without visible growth. Detectable growth occurred at 0.05% oxygen, and 33% of normal colony size under air (21% oxygen) was obtained at 0.15% oxygen. Population selection did not determine survival and growth, but carbon dioxide was required. Characteristic colony morphologies were not evident at the lower oxygen concentrations. Colonial variation was not influenced during survival under anaerobic conditions or growth under low oxygen levels (0.15%). Medium differences were not significant affectors. We concluded that N. gonorrhoeae will grow under tensions suitable for anaerobes, and will demonstrate certain modifications of behavior under these conditions.

Anaerobiosis

Anaerobic survival of clinical isolates and laboratory strains of Neisseria gonorrhoea: use in transfer and storage.

Eleven laboratory strains and 67 clinical isolates of Neisseria gonorrhoeae were tested for the ability to survive during anaerobic incubation. The survival of the laboratory strains was dependent on auxotype, temperature, and cell density on agar plates. For both the laboratory strains and the clinical isolates, anaerobic survival was better at lower temperatures. We concluded that anaerobic incubation, for as long as 7 days, is useful when transporting or storing N. gonorrhoeae.

Anaerobiosis

Metabolic analysis of serologically defined Neisseria meningitidis isolates by frequency-pulsed electron capture gas-liquid chromatography.

Eight serogroups [A, B, C, D, X, Y, 29E [Z'], and W135] along with nongroupable and nontypable strains of Neisseria meningitidis were cultured in a defined liquid medium. The whole-culture spent medium was extracted, derivatized, and analyzed by frequency-pulsed electron capture gas-liquid chromatography. The frequency-pulsed electron capture gas-liquid chromatography profiles were then used to group the organisms on the basis of their metabolic profiles. First, two basic groups were formed which consisted of elevated spent-medium compounds and reduced spent-medium compounds. Then, these two basic groups were further subdivided on the basis of metabolites and cellular fatty acids detected in the spent medium to form a total of seven metabolic groups. The nongroupable strains (with the exception of two strains) fell within one of these metabolic groups. The elevated spent-medium compounds group contained all of the A, B, Y, and Z' strains, and the reduced spent-medium compounds group contained most of the C and all of the W135 and D strains. The potential usefulness of the metabolic group scheme is discussed.

Chromatography, Gas

Rapid differentiation of the major causative agents of bacterial meningitis by use of frequency-pulsed electron capture gas-liquid chromatograph: analysis of acids.

The major causative agents of bacterial meningitis, Haemophilus influenzae serogroup B, Neisseria meningitidis serogroups B and C, Klebsiella pneumoniae, Streptococcus pneumoniae, and two types of Escherichia coli, were cultured in a modified chemically defined Catlin medium and in a commercial version of the unmodified Catlin medium. The spent media were extracted under acidic conditions, and electron-capturing derivatives were prepared by derivatization with trichloroethanol or haptafluorobutyric anhydride. The derivatives were analyzed on a gas chromatograph equipped with a frequency-pulsed electron capture detector and a PEP-2 computer. The data obtained from the study show that these organisms can be easily distinguished from each other on the basis of metabolic products detected in either type of medium. Three different metabolic groups were detected within two serogroups of N. meningitidis. The methods are practical, and the new technique should offer clinical laboratories and hospitals a better method for rapid identification of this important group of pathogens.

Acids

Rapid differentiation of the major causative agents of bacterial meningitis by use of frequency-pulsed electron capture gas-liquid chromatography: analysis of amines.

The major causative agents of bacterial meningitis (Haemophilus influenzae serogroup B, Neisseria meningitidis serogroups B and C, Klebsiella pneumoniae, Steptococcus pneumoniae, and two types of Escherichia coli) were cultured in a chemically defined medium, and selected strains were further studied in Todd-Hewitt medium. After acidic extraction of the spent media with chloroform, a basic extraction was made with chloroform to obtain amines. A third extraction was performed on re-acidified Todd-Hewitt medium with ethyl ether to obtain hydroxyacids. The extracts were derivatized with heptafluorobutyric anhydride-ethanol to form electron-capturing derivatives, and the derivatives were analyzed on a frequency-pulsed electron capture gas-liquid chromatograph (FPEC-GLC) equipped with a PEP-2 computer. The data obtained from the study showed that amines were produced by these organisms that formed characteristic patterns. Different serotypes of K. pneumoniae and the two serogroups of N. meningitidis produced different types of FPEC-GLC profiles within serotypes. E. coli produced several hydroxy acids on Todd-Hewitt medium that made it unique among the organisms studied. The methods used are practical and the techniques have potential for use in clinical laboratories and hospitals as a valuable aid for the rapid identification of the major causative agents of bacterial meningitis.

Amines

[Growth patterns of Neisseria gonorrhoeae: occurrence and significance].

The introduction of transparent mediums for the growth of the gonococcus in the laboratory led to a definitive examination of colonial morphology. Characteristics, such as size, elevation, colour, highlights, opacity,. consistency and evaluation of edges make possible to distinguish five colonial types: T1-T5. The most significant characteristic associated with colony morphology has been virulence; T1/T2-colonies are virulent for humans and T3/T4-colonies are avirulent. Interactions with host cells, metabolic, antigenic and genetic characteristics of N. gonorrhoease of different colonial morphology are discussed.

Animals

Electron capture gas-liquid chromatographic-mass spectral identification of acids produced by Neisseria meningitidis in a defined medium.

The acid metabolites and the cellular fatty acids of three strains of Neisseria meningitidis grown in a chemically defined liquid medium were determined with computerized frequency-pulsed electron capture gas-liquid chromatography. Five acids not previously reported were subsequently identified: isobutyric, octanoic, decenoic (C10:1), dodecenoic (C12:1), and tetradecenoic (C14:1). These acids were produced during active metabolism and were not detected as cellular constituents. The frequency-pulsed electron capture gas-liquid chromatography methods which we used provide a rapid, reliable, sensitive means of detecting both these and other metabolic and cellular acids in spent culture medium.

Chromatography, Gas

Identification of some basic extractable compounds produced by Neisseria gonorrhoeae and Neisseria meningitidis in a defined medium.

Electron capture gas-liquid chromatography and mass spectrometry have been used to identify some of the basic extractable heptafluorobutyric anhydride reactive compounds found in a defined medium after 20 h of growth by Neisseria gonorrhoeae and N. meningitidis. Acetoin, 2,3-butanediol, pyrroline, and 1,3-diaminopropane were identified by both gas chromatography and mass spectrometry; 2-hydroxy-pyrrolidine and 3-aminomethyl-pyrrolidine were tentatively identified by mass spectrometry. A possible origin of the amines is through enzymatic oxidation of polyamines.

Acetoin

Treatment of gonorrhea with trimethoprim-sulfamethoxazole.

The following regimens were randomly administered to 271 men with gonococcal urethritis: 4.8 X 10(6) units of aqueous procaine penicillin G intramuscularly plus 1 g of probenecid orally (APPG); nine tablets of trimethoprim-sulfamethoxazole (TMP-SMZ; 720 mg of TMP and 3,600 mg of SMZ), orally as a single dose (TMP-SMZ-9); and 12 tablets of TMP-SMZ (960 mg of TMP and 4,800 mg of SMZ) orally as two doses of six tablets taken at a 6-hr interval (TMP-SMZ-12). The failure rates of the APPG, TMP-SMZ-9, and TMP-SMZ-12 regimens were 4%, 23%, and 19%, respectively. APPG was significantly more effective (P less than 0.05) than TMP-SMZ-9 or TMP-SMZ-12. Isolates of Neisseria gonorrhoeae from treatment failures as compared to those from treatment successes were significantly more resistant to SMZ (P less than 0.01) and to the TMP-SMZ combination in a ratio of 19 parts SMZ to one part TMP (P less than 0.05). Minimal inhibitory concentrations of SMZ, TMP, TMP-SMZ, and penicillin G showed positive correlation coefficients.

Administration, Oral

Identification of Neisseria by electron capture gas-liquid chromatography of metabolites in a chemically defined growth medium.

A dual-purpose study was carried out in an attempt to develop a rapid, sensitive method to identify Neisseria species by gas chromatography and to learn more about the metabolism of these organisms. Sixty-nine isolates of Neisseria were grown in a chemically defined fluid medium; the spent medium was extracted sequentially at pH 2 with diethyl ether and at pH 10 with chloroform. The pH 10 extracts were derivatized with heptafluorobutyric anhydride and analyzed by electron capture gas-liquid chromatography. The resulting spent culture medium electron capture gas-liquid chromatography profiles showed several qualitative and significant quantitative differences among the Neisseria species potentially useful in separating and identifying these organisms. Putrescine and cadaverine which were present in the spent culture medium of some Neisseria, including N. gonorrhoeae, were tentatively identified. Substituting carbohydrates for the chemically defined medium containing glucose in the base medium produced altered profiles with increased quantitative and qualitative differences.

Amines

Electron capture gas chromatographic detection of acethylmethylcarbinol produced by neisseria gonorrhoeae.

Acetylmethylcarbinol (acetoin) production by Neisseria gonorrhoeae and other Neisseria species was established by gas-liquid chromatography and by mass spectrometric data. Sixty-nine isolates of Neisseria were tested by incubating them in a chemically defined fluid medium. The medium was extracted with organic solvents and derivatized with heptafluorobutryic anhydride for gas chromatography and mass spectrometry. Cultures of 58 of the same strains were tested with the conventional Voges-Proskauer reagents, and results were compared with those of gas-liquid chromatography. When glucose was used as an energy source, N. gonorrhoeae, some N. meningitidis, and N. lactamica produced enough acetoin in 16 h to be detectable by either method, whereas other Neisseria species produce amounts detectable only by gas chromatography. The conventional acetylmethylcarbinol test with the chemically defined medium and maltose as an energy source might be used to develop methods that would differentiate certain members of the genus, including the pathogenic species.

Butanones

New developments in the laboratory diagnosis of gonorrhea.

Various laboratory technologies have been developed to aid in diagnosing gonorrhea. Transport of viable specimens to the laboratory and the cultivation of N. gonorrhoeae on selective media are among the most effective diagnostic procedures in clinical microbiology. Several sensitive serologic procedures have been developed for detecting antibodies to N. gonorrhoeae.

Antibodies, Bacterial

Rapid fermentation confirmation of Neisseria gonorrhoeae.

A rapid sugar fermentation procedure has been developed for the confirmation of Neisseria gonorrhoeae from either primary isolation media or purification media. A lightly buffered salt solution with pH indicator and sugars was heavily inoculated with presumptively positive growth. Proper fermentation patterns are obtained in 1 to 4 h with no interference from inhibited contaminants or variation in results due to differing growth requirements of gonococcal strains.

Bacteriological Techniques