Application of Electron Microscopy and Energy Dispersive X-Ray Spectroscopy in the Characterization of Rhodomicrobium vannielii
Abstract:Bacterial species Rhodomicrobium vannielii (UKMP4-5A), isolated from a hot spring in Malaysia was grown in Malate Yeast Extract (MYE) broth at 47 °C with 2000 lux of continuous lighting and optimal pH of 7.3. In low light of less than 2 kilo-lux, the organism drifted towards light by attaching to the walls of culture bottles while older cells formed debris of interwoven brownish cell deposit at the base of broth media. Transmission electron microscopy (TEM) of negatively stained cells revealed lemon-ovoid shaped multiple peritrichously flagellated young cells with vegetative polar proliferation by budding. Flagella were missing in most full-grown cells with size-range of 1.0–1.9 m by 1.8–3.1 m. Young daughter-cells emerged from budding ends of vegetative hyphae of adult cells. Formation of hyphae was observed after few hours of incubation of sub-cultured broth with highly motile multi-flagellated young bacterial cells. The formation of dense septa preceded the outbreak of young swarmer cells. The dense septa were an indication of maturity of grown cells, observed in both negatively stained cells and in ultrathin sections. Irrespective of denseseptal formation patterns, multi-tubular and twin-nodular growths with exospores were observed in some of the cells. This was the first ever reported finding of the twin-nodular growth. Ultrathin microtomy revealed the presence of lamellae and other intracellular inclusions. Anatomic inclusions in filamented cells were less diversified than those in single non-filamentous. Bacterial identity of the organism was reaffirmed with energy dispersive X-ray spectroscopy (EDS) with min-max ratio of weight percentage of normalised analysis detected at 68 to 75% of carbon along with other elements.
Document Type: Research Article
Publication date: February 1, 2011
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