Date of Award
Master of Science (MS)
A microorganism resembling a fungus, as revealed by light microscopy, was collected from a preserved cat dissection specimen in the Fort Hays State University anatomy laboratory. The organism subsequently was isolated and grown in pure culture on tryptic soy agar. Fatty acid methyl ester (FAME) analysis, scanning electron microscopy, partial 16S rRNA sequencing, and full gene 16S rRNA sequencing were used to partially characterize the organism. MIDI Labs (Newark, Delaware) sequenced and analyzed the first 500 base pairs of the 1,422 base pair 16S rRNA gene. The 16S rRNA gene was then sequenced at Fort Hays State University (FHSU) and was re-submitted to MIDI Labs for the full 16S rRNA gene sequence. The sequence obtained at FHSU and the full 16S rRNA gene sequence obtained from MIDl Labs was compared to all sequences in the GenBank database. The nucleotide sequence data collected by MIDI Labs, from both the partial and the full gene sequence analysis, and the first 500 base pairs of the gene sequence data obtained at FHSU were identical. Initially, the partial 16S rRNA sequence data indicated that this organism was most likely a new species within the genus Streptomyces. However, the full gene sequence more accurately revealed that this organism might be a new species within the genus Saccharopolyspora. The FAME analysis performed by MIDI Labs indicated that this organism might belong to the genus Corynebacterium. Light microscopy revealed that this organism is a Gram-positive filamentous bacterium. Scanning electron microscopy allowed the determination of the fine detail of cellular morphology of this organism including the ability to sporulate. FAME analysis and 165 rRNA sequence analysis demonstrated that this micro-organism could be a previously undescribed species within known genera. This organism was shown to be a close relative of Saccharopolyspora hirsute strain ATCC 27875. Saccharopolyspora species are Gram-positive, spore forming filamentous bacteria. The Gram reaction for this organism is Gram-positive, the scanning electron micrographs indicate filamentous hyphal growth and the potential to sporulate, the cellular fatty acids are consistent with those expected for members of the genus Saccharopolyspora, along with the sequencing data all of the results obtained indicate that the organism can be accurately identified as belonging to the genus Saccharopolyspora.
Copyright 2005 Sherrie Stawinski
Stawinski, Sherrie, "Characterization of a Saccharopolyspora by 16S Rrna Sequencing, Fatty Acid Methyl Ester Analysis, and Scanning Electron Microscopy" (2005). Master's Theses. 2962.