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Isolation of Azotobacter
Transcript of Isolation of Azotobacter
Aerobic Gram-Negative Organism
Large and ovoid morphology
Fix atmospheric Nitrogen
Do not have symbiotic relations
Free-living in soil and water
Distinguished by pigmentation
Two samples of soil were collected
To confine and determine species from
by using soil samples, Mannitol Salts enrichment media, and multiple identification tests .
and Biofilm Project
Prepared Mannitol Salts Broth (N-free)
Niki Yazdani and Lauren McGehee
Mannitol 10.5 g
K2HPO4 .458 g
MgSO4 .14 g
CaSO4 .09 g
NaCl .14 g
CaCO3 3.5 g
dH2O 1000 mL
Motility deep for our four colonies
All four colonies tested positive
Wet mounts also positive
Used Mannitol test to see if the colonies are able to ferment Mannitol as a carbon source
All tubes changed from red to yellow indicating a positive test
Oxidase test was used to determine if ATP is generated by cytochromes
Oxidase reagent was combined with water
Colonies were placed on reagent
Positive result for all colonies
Table of Results
We narrowed down our colonies to 3 species:
Since the test results are similar, we couldn't distinguish between A. armeniacus and A. chroococcum
Biofilm grown from deionized water and soil samples
Sample sat for a number of days
LSL to EMB
Part 2: Biofilm Isolation
API 20E & 20NE
API 20E: Enterics = lactose fermenters
API 20NE: Nonenterics = non fermenters
Rapid ID Results
API 20 NE
Opaque and convex morphology
Randomly placed granules
Mannitol and caproate as carbon source
Many carbon sources
Easily cultured and grown
Part 3: Rapid Identification Systems
Bergey, David H., and John Holt. Bergey's Manual of
Determinative Bacteriology. Philadelphia: Lippincott, 2000. 552-72. Print.
Identified three possible species of
Better identification if we did more tests
More conclusive results
If we didn't have snow days
cells." 2009. JPEG file.
"Biofilm Formation." 2011. JPEG file.
in the phase microscope at 400X."
2007. JPEG file.
" Tchan & New (1984) in
Bergey's Manual of Systematic Bacteriology, vol. 1, N.R.
"Rough and smooth colony types of
hours." 2005. JPEG file.
EMB." 2006. JPEG file.