Microbes in Soil

Alexis Parsons, Lianna Fiore, Michael Hicks, Terry Hicks

Department of Biology and Biomedical Engineering, Rose-Hulman Institute of Technology

Objective Biochemical Testing Results Conclusions

• Extract/Isolate microorganism from Rose-
Hulman soil samples
• Determine if any of the soil isolates Michael: Pseudomonas,
produce antibiotics Moraxella, or Pasteurella
• Sequence and identify soil isolates
Alexis: Enterococcus
species
Lev EMB test was MacConkey Test 5% Blood Agar 5% Blood
Materials and Methods negative for was negative for test was Agar test was
lactose lactose negative for positive for α- Lianna: Enterococcus
fermentation hemolysis hemolysis species, Shigella species

Terry: Staphylococcus
Serial Dilutions Grid Plate, front and back species
After collecting soil samples, we
performed serial dilutions. Then we made grid
plates to begin testing each isolate. To test the
isolates for antibiotic resistance, we used Mannitol Salt Gram Stain Positive indole Motility test was
Test was was positive test
patch plates, turtle track plates, and safe negative and
positive
ESKAPE pathogens relatives. shows anerobic
growth
References
Laboratory Manual used as Experimental
PCR and Gel Electrophoresis Guide: Hernandez, Simon, et al. Tiny Earth.
In addition to biochemical testing, the XanEdu Publishing Inc, 2018.
polymer chain reaction (PCR) was run
Patch Plate Turtle Track Plate Assay Extract Plate as a means of amplifying the isolates'
Carbohydrate genetic material. Using gel
After identifying potential antibiotic producing agents, electrophoresis to gauge the success of
we extracted the isolated using lawn plates and then Fermentation Test
was positive for the reaction, this process was found to
using sterile water and ethyl acetate to extract the
antibiotics. We tested the antibiotics using an assay
Dextrose/Durham have failed in DNA amplification. Acknowledgements
sugars
extract. We would like to thank Dr. Jen
Positive Simmon's Sanger Sequencing O'Conner, the lab technicians, and Tiny
We also characterized our antibiotic producing Citrate Test Earth for setting up this experiment
indicating the ability
Had the PCR been successful, the
isolates using Gram staining, Oxidase Test, Catalase amplified DNA of the isolates would have and helping us learn new techniques for
of the testing our isolates for antibiotic
Test, Blood Agar Test, MacConkey Test, Lev EMB microorganism to been sequenced through the Sanger
test, TSI Agar Test, Simmons Citrate Test, MR-VP Sequencing procedure, allowing properties.
use citrate as a
Broth, SIM and Indole Tests, Carbohydrate primary carbon comparisons to database genomes.
Fermentation Test, and Mannitol Salt Test. source.