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Pseudomonas Lab Report

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Pseudomonas Lab Report
Pseudomonas taiwanensis – Enrichment, Isolation and Identification
Microbiology 521
2/10/12

Purpose: The purpose of this experiment was to enrich Pseudomonas bacteria, isolating a species of Pseudomonas and identifying it using phenotypic properties and DNA sequencing as an existent or completely new and undiscovered species of Pseudomonas.

Overview: Genus Pseudomonas is a chemoheterotrophic bacteria found in soil and water. They are Gram negative, motile, paired rods that are also oxidase-positive. Pseudomonas species are well known for their ability to metabolize many different organic nutrients, allowing them to be viable in multiple, diverse conditions. The Pseudomonas organisms had to initially be enriched from a soil sample
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The TSA plate was tested for α-amylase production but flooding the plate with iodine and a clear zone did not form around the streak, meaning that the organism could not use the starch and therefore no starch utilization. The Pseudomonas-F agar was viewed in the dark under UV light and fluoresced a yellow pigment, meaning production of pyoverdin and a positive on fluorescent pigmentation. The organism was capable of lipase production as the streak had a clear zone around it in the spirit blue agar, meaning the lipase reagent in the medium was degraded and utilized. The gelatinase test showed no clearing on the strip, meaning no gelatinase utilization. The denitrification was tested and no result showed positive. Reduction of nitrate did not occur as no pink substance was observed after addition of 12 drops if sulfanilic acid solution. A red color did not appear during a dropwise addition of α-napthylamine solution and a pink color formed when zinc was added, meaning the nitrate was still present in the medium. The anaerobic plate showed growth, meaning that the organism was a NO3- electron acceptor. Trehalose utilization did not occur but histidine, glucose and arginine were all utilized by the organism. Finally, visible pigmentation was observed on plates. The PCR amplification ran smoothly and 2 100% matches were found in the RDP for the sequence of the organism amplified. These were P. taiwanensis and P.

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