Isolation and Enrichment of soil bacteria Regina Gohl

Isolation and Enrichment of soil bacteria Regina Gohl, K. Noreen Boyle 02/11/2011 University of Oldenburg - ICBM - Practical course Physiology of bacteria (Group B 1) – final presentation

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 2 Introduction • 1 soil probe 2 experiments Isolation Dilution series Introduction Enrichment ? Aims & Expectations Medium variation Course results 37 °C Pasteurisation Nitrogen + yeast Salt+yeast Acidic Alkaline Summary

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 3 Aims and expectations • • Cultivation of soil bacteria Determination of abundance Comparison: fast vs abundant Influence of different media Expectations Direct isolation Enrichment Types of bacteria several one - few Colony size small large Growth slow fast Conclusion abundant fast growing Introduction Aims & Expectations Course results Summary

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 4 Direct Isolation Cells/g soil ~ 10^9 bacteria/g soil 3. 1 x 10^6 5. 4 x 10^6 3. 3 x 10^6 2. 8 x 10^6 cells/g soil 1. 9 x 10^6 4. 7 x 10^6 1. 9 x 10^6 ~1 % of bacteria can be cultured in the lab 1. 0 x 10^6 0. 8 x 10^6 Less bacteria than expected Introduction Aims & Expectations Course results Summary

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 5 Direct Isolation Introduction Aims & Expectations Course results Summary

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 6 Enrichment Treatment Oxidase test Gram test Catalase test Increased temperature + - - - Pasteurisation + - - - + + - nitrogen source - + + -nitrogen source + yeast extract + + + + salt + yeast extract + + + acidic + + + alkaline + + + http: //www. laborwissen. de/wiki/ tiki-index. php? page=Katalase-Test Introduction Aims & Expectations Course results Summary

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 7 Enrichment Treatment Motility Growth Size Shape mobile fast large round, beige Pasteurisation immobile slow small round, beige - nitrogen source immobile, rods slow small white, smooth - nitrogen source + yeast extract mobile fast large yellow, smooth + salt - slow small, round, white + salt + yeast extract - fast large, round, white acidic cocci, immobile slow small beige, bright alkaline cocci, immobile slow small beige, bright Aims & Expectations Increased temperature Introduction Course results Summary

Regina Gohl, K. Noreen Boyle ICBM Uni Oldenburg 8 Summary • Results as expected • Variety of bacteria in soil • Different treatment growth / no growth - nitrogen source + yeast extract Introduction Aims & Expectations - nitrogen source Course results Summary

Thank you for your attention