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Microbiology Unknowns

Microbiology Unknowns Stephanie Everitt, Carly McMacken, Quiana Neilson, & Heather Peterson BIOL 201C – Spring 2008 OVERVIEW Introduction Inoculation Introduction to unknowns Identification G+ species G- species Conclusion Best Guess Future Research

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Microbiology Unknowns

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  1. Microbiology Unknowns Stephanie Everitt, Carly McMacken, Quiana Neilson, & Heather Peterson BIOL 201C – Spring 2008

  2. OVERVIEW • Introduction • Inoculation • Introduction to unknowns • Identification • G+ species • G- species • Conclusion • Best Guess • Future Research Image Source: Missouri University of Science and Technology; http://web.mst.edu/~microbio/Bio221.html

  3. INTRODUCTION

  4. Specimen Collection • Sample source: • tree bark on campus Image Source: Microshield Environmental Services; http://www.microshield-es.com/airqualitymoldtesting.html

  5. Inoculation • Both colonies: • Small • Circular Beige colonies (white) Orange colonies Figure 1. Culture of tree sample on TSA; incubated at 37° for 24 hrs. • Smooth edges • Convex elevation • Mucoid consistency

  6. Unknown Orange • G+ • Rod shape • Asporical • Random arrangement • ~2μm Figure 2. Gram Stain of unknown orange bark sample at 1000x.

  7. Unknown White • G- • Slender rod shape • Asporical • Random arrangement • ~2 μm Figure 4. Gram Stain of unknown white bark sample at 1000x.

  8. IDENTIFICATION

  9. G- : Unknown White • Tests Positive for: • Growth in high salinity of bile salts • Hydolysis of: • Casein • Gelatin (anaerobically) Figure 5. Gram Stain of unknown orange bark sample at 1000x using a zoom lens camera. The black on the right is the microscope pointer.

  10. G- : Unknown White • Tests negative for: • Fermentation of: manitol, lactose, glucose, maltose, and sucrose • Hemolysis • Citric acid as a C source • Indole production • N reduction • Mixed acid & 2,3-butanediol fermentation Figure 5. Gram Stain of unknown orange bark sample at 1000x using a zoom lens camera. The black on the right is the microscope pointer.

  11. G- : Unknown White • Most similar to Pseudomonas aeruginosa • Eliminating test • Milk Nutrient Agar Figure 6. Pseudomonas aeruginosa on Pseudomonas Isolation Agar has a similar appearance to its growth on Milk Nutrient Agar. Image Source: The Environmental Reporter; http://www.emlab.com/s/sampling/env-report-03-2007.html.

  12. G- : Unknown White • Is it actually G+? • Same test results on average: • 2.9 – G- • 3.6 – G+ • Pallisades • Corynebacterium • G+ rods • pallisades Figure 5. Gram Stain of unknown orange bark sample at 1000x using a zoom lens camera. The black on the right is the microscope pointer.

  13. G- : Unknown White • Is it actually G+? • Most conclusive cell wall test? • Antibiotic results Image Source: The Microbial World; http://www.biology.ed.ac.uk/research/groups/jdeacon/microbes/penicill.htm

  14. G-: Unknown White Table 1. Antibiotic clearing results (mm) for Unknown White, G+ known species, and G- known species. * indicates resistant strains present within the clearing.

  15. G- : Unknown White • Other Pseudomonas? • Non-fermenters • Only 15% of G-, 70% Pseudomonas • Yes • Unknown white has high salinity tolerance •  No! Figure 5. Gram Stain of unknown orange bark sample at 1000x using a zoom lens camera. The black on the right is the microscope pointer.

  16. G- : Unknown White • Pseudomonadaceae • Frateuria • if tests + for acidity tolerance Figure 5. Gram Stain of unknown orange bark sample at 1000x using a zoom lens camera. The black on the right is the microscope pointer.

  17. Unknown Orange Enrichment Cultures • Can grow in high salinity • Tests negative for: • Fermentation of: manitol, lactose, glucose, maltose, and sucrose • Hydolysis of: starch, casein, gelatin, and urea • Hemolysis Figure 3. Gram Stain of unknown orange bark sample at 1000x using a zoom lens camera.

  18. CONCLUSION

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