Isolation and Characterization of Bacteria Purified From Tropical Puerto Rican SoilsFRANCISCO J. RODRÍGUEZ JIMÉNEZ

JARIEL Y. RAMÍREZ VIRELLABiol 3955 – Biomedical Techniques

Dr. Díaz, Dr. Rubin & Dr. GonzálezMay 15, 2015

Introduction

• Bacteria in worldwide commerce.

•Antibiotics have boosted health worldwide (Davies 2010)

• Sepsis = worsening (Uppu 2015)

•Isolating and culturing = maybe new substance/strain

Materials and MethodsSoil Extraction

Soil A: Taken February 29, 2015; Barrio Frank Rodriguez, Cayey, Puerto Rico.

Soil B: Taken February 25, 2015; Laguna de Tortuguero, at Vega Baja, Puerto

Rico; Stored at 4⁰C

Materials and MethodsSoil A: Taken February 29, 2015 Soil B: Taken February 25, 2015

Materials and Methods1. R2A & TSA

2. Two colonies were chosen by qualitative factors.

3. Selected colonies were isolated and purified

4. Gram Stain & Microscopy

5. PCR/Electrophoresis

6. Antibiotic production

ResultsSOIL A: S15UPRC-RISEFRJ30R01A

oYellow-toned

oGelatinous and sticky

oSeemed to absorb nutrients at a fast rate

oPh 6.5

oGram positive

SOIL B: S15UPRC-RISEJYRV30R01A

oWhite

oExtensions to exterior (like Fimbriae)

oPh 5.5

oGram positive

Results Antibiotic Porduction: PCR/Electrophoresis

Conclusion•R2A-plate nutrients are best

•Cell wall, rod-shaped

•No antibiotic production… Changed plates?

•Negative results in PCR… human error?

Cited Literature Davies J & Davies D. 2010. Origins and Evolution of Antibiotic Resistance. Microbiol Mol Biol Rev. Volume XX. 74(3): 417–433.

Available from: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2937522/ doi: 10.1128/MMBR.00016-10

Ling LL, Schneider T, Peoples AJ, Spoering AL, Engels I, Conlon BP, Schaberle TF, Hughes DE, Epstein S, Jones M, Lazarides L, et. al. 2015. A new antibiotic kills pathogens without detectable resistance. Nature, Volume 517. Jan; 455-459. Available from: http://www.nature.com/nature/journal/v517/n7535/full/nature14098.html doi: doi:10.1038/nature14098

Nichols D, Cahoon N, Trakhtenberg NE, Pham L, Mehta A, Belanger A, Kanigan T, Lewis K, Epstein SS. 2010 Apr. Use of Ichip for High-Throughput In Situ Cultivation of “Uncultivable” Microbial Species. Appl Environ Microbiol. 76(8): 2445–2450. Available from: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2849220/ doi: 10.1128/AEM.01754-09

Penhale S. 2011. Using Scientific Literature in Biology Courses. Earlham College. Available from: http://www.earlham.edu/media/170978/biocite.pdf

Rodriguez JC, Pino C, Tauch A, Murcia MI. 2015. Complete Genome Sequence of the Clinical Beijing-Like Strain Mycobacterium tuberculosis 323 Using the PacBio Real-Time Sequencing Platform. Genome Announc, 3(2):e00371-15. doi:10.1128/genomeA.00371-15

Suzuki S, Horinouchi T, Furusawa C. 2015. Suppression of antibiotic resistance acquisition by combined use of antibiotics. Journal of Bioscience and Bioengineering, Volume XX. 1-3. Available from: http://www.researchgate.net/publication/273326671_Suppression_of_antibiotic_resistance_acquisition_by_combined_use_of_antibiotics doi: 10.1016/j.jbiosc.2015.02.003

Uppu DSSM, Ghosh C, Haldar J. 2015. Surviving sepsis in the era of antibiotic resistance: Are there any alternative approaches to antibiotic therapy?. Microbial Pathogenesis, Volume 80. 7-13. Available from: http://www.sciencedirect.com/science/article/pii/S0882401015000121 doi:10.1016/j.micpath.2015.02.001