Nocardia, Mycobacteria, Corynebacteria
John A Luccivisibility
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6 pages
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In contrast to the complex branching of Streptomyces sp., the acid-fast Corynebacterineae genera Nocardia, Mycobacteria and Corynebacteria are all irregularly branched with varying amounts of filamentous morphology. Enrichment was partaken using soil to inoculate broths and subsequent respective plates of n-amyl alcohol, cyclohexanol, phenol and pyridine. 1 gram of soil each was used to inoculate differential media comprised of mineral salts and either 0.3 % N-amyl alcohol, 0.5 % Cyclohexanol, 0.1 % Phenol, or 0.1 % Pyridine. Media was then incubated without shaking in a 30C and then streaked onto mineral salts agar with homologous substrates N-amyl alcohol, Cyclohexanol, Phenol, and Pyridine and again incubated at 30C. The macroscopic and microscopic confliction of the observation of the cells grown on phenol, which macroscopically appeared as slow growing, largely translucent colonies. Under the microscope, however, extensive branching appeared, with duel branches formed outward at the same point longitudinally along a central stalk, subsequently fanning out into greater subdivisions, easily observed at the 1000x as fanning out in series of 6, with three larger cells of club rod morphology or possibly lophotrichously flagellated. The cells in cyclohexanol exhibited similar repeated-ray morphology, seemingly in series of fives with much greater diversity of morphology, with palisades of helical folds surrounded by a diplobacillal majority. The cells grown in media containing N-amyl alcohol showed straight to slightly curved rods with irregularly individual proximal granules. Pyridine appeared brown after two days but transformed over two weeks into compact white spores which were pigmented on cortical sections. Phenol
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In contrast to the complex branching of Streptomyces sp., the acid-fast Corynebacterineae
genera Nocardia, Mycobacteria and Corynebacteria are all irregularly branched with varying amounts of
filamentous morphology. Enrichment was partaken using soil to inoculate broths and subsequent
respective plates of n-amyl alcohol, cyclohexanol, phenol and pyridine.
1 gram of soil each was used to inoculate differential media comprised
of mineral salts and either 0.3 % N-amyl alcohol, 0.5 % Cyclohexanol, 0.1 % Phenol, or 0.1 % Pyridine.
Media was then incubated without shaking in a 30C and then streaked onto mineral salts agar with
homologous substrates N-amyl alcohol, Cyclohexanol, Phenol, and Pyridine and again incubated at 30C.
Phenol
The macroscopic and microscopic confliction of the observation of
the cells grown on phenol, which macroscopically appeared as slow growing, largely translucent
colonies. Under the microscope, however, extensive branching appeared, with duel branches formed
outward at the same point longitudinally along a central stalk, subsequently fanning out into greater
subdivisions, easily observed at the 1000x as fanning out in series of 6, with three larger cells of club rod
morphology or possibly lophotrichously flagellated. The cells in cyclohexanol exhibited similar repeatedray morphology, seemingly in series of fives with much greater diversity of morphology, with palisades
of helical folds surrounded by a diplobacillal majority. The cells grown in media containing N-amyl
alcohol showed straight to slightly curved rods with irregularly individual proximal granules. Pyridine
appeared brown after two days but transformed over two weeks into compact white spores which were
pigmented on cortical sections.
N-amyl alcohol
Cyclo hexanol
Pyridine
Soil Extract with Glycerol
Soil extract
Results at the primary point were indicative of Corynebacteria, Mycobacterium and a mixed culture of
Corynebacteria and Nocardia, cultured on phenol, cyclohexanol, and n-amyl alcohol, respectively. The
morphology of the pyridine growth was less obvious, thus, further isolation was partaken using soil extract
with and without glycerol, as well as yeast-glucose agar. Colonies formed on the yeast-glucose agar were
a vibrant shade of yellow, comparable to myxococcus. Microscopically, the cells comprised of paired
diploid rods and a strange filamentous form. Cells on the soil extract were club shaped appearing
moderately filamented at low resolution, with colonies that were mucoid, yet bumpy and rough. The soil
extract with glycerol isolated organisms quite homologous in morphology, forming chained series of rods
which appeared to aggregate to a significant amount. The colonies on the soil extract with glycerol were
strange; extremely hard and rugged, almost impossible to loop off. The colonies also almost gave off a
luminescence, appearing shiny in the light, observable in the photo above as diffraction of the flash.
Nocardia is specifically visualized as irregularly branching diptheroid cells, containing
arthrospores, with colonies wrinkled and nodular with a dull waxy surface. Organisms containing similar
morphology were observed on the culture grown on soil extract. A separate species of Nocardia,
comprised of wrinkled white colonies can be observed on the plate containing soil extract with glycerol.
The specific species of these two isolations could possibly be Nocardia pulmonalis, described as having
vegetative mycelium that is hard and yellow, with white aerial mycelium, with substrate hyphae that
divide into chains of acid-fast cocci, and Nocardia transvalensis, indicated by non-acid-fast diastatic
organisms with weakly refractive cells, respectively. The glucose-yeast extract yielded organisms that
formed series of palisades with metachromatic granules representing stored phosphate. This is clearly
indicative of Corynebacteria species, possibly Corynebacterium kutscheri listed in Bergey’s ith olonies
o ser a le as Small, thin, yellowish white, translucent, serrate. Regardless of species, it could easily be
stated that the mammalian pathogenesis caused by many members of the suborder Corynebacterineae,
has garnished a reputation of simplistic, yet deadly, but the diverse series of species observed from the
soil sample gives evidence to their presence in soil, either as pathogenic or commensal to other organisms.
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