Microbiology - Volume 136, Issue 6, 1990

Summary: An anaerobic enrichment from lake mud yielded a pure culture of a facultatively anaerobic bacterium able to grow on malonate under strictly anaerobic conditions. Strain 16mall was identified as a member of the family Enterobacteriaceae, and assigned to the genus Citrobacter on the basis of morphological, metabolic and biochemical characteristics. Malonate was fermented under strictly anaerobic (sulphide-reduced) conditions to acetate and CO2 concomitant with growth. A maximum growth rate of 1.88 generations h−1 (μ = 1.30 h−1) was measured. The dry weight yield of cells from malonate was estimated at 2.5 g mol−1. Yeast extract was required for growth on malonate: other additives, or a vitamin solution, could not replace this requirement. Other dicarboxylic acids were not degraded in the absence or presence of malonate. Malonate was degraded under anaerobic, but not aerobic conditions. Malonate-decarboxylating activity was inducible by malonate under both anaerobic and aerobic conditions, and was not expressed in glucose- or citrate-grown anaerobic cultures. Monensin had no effect on malonate degradation, while 2,4-dinitrophenol decreased the rate of malonate degradation. This, with the lack of a sodium requirement for anaerobic growth on malonate, suggested that ATP generation may not be mediated by a sodium-pumping mechanism.

Summary: Lucifer yellow (LY), an impermeable fluorescent dye used as a marker for fluid phase endocytosis, was internalized by Candida albicans. As observed by fluorescence microscopy, incubation of C. albicans with LY in potassium phosphate buffer (pH 6.0) and glucose (2%, w/v) resulted in localization of the dye inside vacuoles. Sodium azide and carbonyl cyanide m-chlorophenylhydrazone, which are inhibitors of energy metabolism, decreased the uptake of the dye. The optimum temperature for uptake was 30.C; no internalization was observed at 0.C. Quantification of cell-associated LY by fluorescence spectrometry showed an uptake linear with time and not saturable over a 400-fold range of concentration. Thus, C. albicans internalized LY into vacuoles by a nonsaturable and time-, temperature- and energy-dependent process consistent with fluid phase endocytosis. Both the yeast and mould phase of this dimorphic fungus endocytosed LY. Growth in complex medium appeared to be required to enable the cells to internalize LY. However, addition of peptone or yeast extract to the phosphate buffer/glucose assay medium interfered with LY uptake by causing an apparent increase of exocytosis. These studies provide the first evidence of fluid phase endocytosis in C. albicans and may explain how some large molecules, such as toxins and cationic proteins, enter C. albicans.

Summary: Hyphal extension in the filamentous actinomycete Streptomyces coelicolor A3(2) was shown to occur by addition of newly synthesized wall material in an apical extension zone. Incubation of mycelia with tritiated N-acetyl-D-glucosamine (GlcNAc), a precursor of peptidoglycan, resulted in localized incorporation of label at the apex, as indicated by light microscopic and electron microscopic autoradiography. Within the hyphal extension zone there was a sharp decrease in incorporation with increasing distance from the apex. Hyphal tip shape, examined by low-temperature scanning electron microscopy, approximated to a semi-ellipsoid of revolution and was not hemispherical. Tip shape could be represented accurately by polynomial equations of degree less than seven. The surface stress theory was successfully applied to hyphal tip growth, with tip shape related qualitatively to the inverse of surface tension within the wall of the extension zone. Surface tension was assumed to be inversely proportional to the rate of incorporation of tritiated GlcNAc. Treatment of surface-grown hyphae with β-lactam antibiotics resulted in localized swelling of hyphal tips. Lysozyme caused swelling of tips and of other regions of hyphae, frequently giving a beaded morphology associated with septa.

Summary: Four, NAD-independent, clinical isolates of Haemophilus parainfluenzae were recovered from a genital ulcer, a purulent skin lesion, a sputum specimen and a throat swab respectively. With the exception of NAD requirement, the strains exhibited the biochemical characteristics of H. parainfluenzae biotype II. The genetic relationship between these isolates and a standard strain of H. parainfluenzae was determined by testing transforming activities of two chromosomal markers, streptomycin resistance and nalidixic acid resistance. The clinical isolates were efficient donors and recipients in transformation. In addition, we demonstrated transfer of the genes conferring NAD independence to typical, NAD-requiring H. parainfluenzae and Haemophilus influenzae strains.

Summary: Total DNAs of 18 strains of Azospirillum from different sources and geographical areas were compared by restriction endonuclease pattern analysis. Fragments obtained with HindIII or BglII were separated by PAGE and stained with silver nitrate. Each strain possessed a unique and reproducible fingerprint with each enzyme, thereby facilitating strain recognition. UPGMA analysis recovered clusters of band patterns that were compared to the distribution of species within the genus Azospirillum.