Isolation and Characterization of Bacteria from Tropical Soils Alejandra Ferrer 1 Nicole S. Rivera 1 Jessica Ortiz 1 Carolina Huertas 1 Dr. Michael Rubin Date: May 15, 2015 RISE Program Biology and Chemistry Department University of Puerto Rico at Cayey 1 This author contributed equally to this project
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Isolation and Characterization of Bacteria from Tropical Soils
Alejandra Ferrer1
Nicole S. Rivera1
Jessica Ortiz1
Carolina Huertas1
Dr. Michael Rubin Date: May 15, 2015
RISE Program Biology and Chemistry DepartmentUniversity of Puerto Rico at Cayey
1 This author contributed equally to this project
Background • Bacteria are prokaryotic microorganisms that can be differentiated by
their peptidoglycan layer located in the cell wall. • This aspect classifies them as gram-positive or gram-negative.
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Antibiotic Era
• The discovery of penicillin in 1928 was by Alexander Fleming.
• Since 1940’s scientists, using Fleming’s research as a source, have been studying and developing antibiotics in order to reduce illness and death from infectious diseases (CDC 2014).
• Rene Dubos- discovered and isolated an antibacterial compound called tyrothricin.
• Recently, scientists have discovered a new antibiotic, from the soil, which name is Teixobactin (Ling et al. 2015).
• Antibiotic resistance is produced when the capability of killing or inhibiting bacterial growth is eliminated. The bacteria can grow in the presence of the antibiotic (APUA 2014).
• In a recent investigation Walsh and colleagues (2003) isolated 412 antibiotic resistant bacteria from agricultural, urban, and pristine soils.
Significance● The importance of this study is that it attempts to find new
bacteria from tropical soils with antibacterial properties.
● This investigation responds to the need for new antibiotics that can inhibit bacterial reproduction.
Problem
• Can we find bacteria in soil that have useful properties such as antibiotic production and antibiotic resistance?
Hypothesis
• It is hypothesized that different properties of the isolated soil bacteria will be characterized such as bacterial structure, antibiotic resistance and antibiotic production .
• There will be more bacteria found in tropical soils capable of antibiotic production and resistance . (Dubisnky et al. 2010)
Goals• Isolation, purification and freezing of soil bacteria
• Gram staining and colony morphology
• Test for antibiotic production and resistance • Generation of genomic DNA, PCR, gel electrophoresis
and purification
• Sequencing and Bioinformatics Analysis
Experimental Design
http://amoya.webnode.es/foros-de-debate/
1 gram of soil
mix
Label the tubes 0→ -5
water (8.5ml)
NaCl(90µl)
streak the plate NaCl
medium plates
collect the colony
II. Purification
III. Cryogenic Freezing
10 mL pipette
S15UPRC-RISEInitials30M01S :Spring15 :2015UPRC: UPR-CayeyRISE: RISE ProgramInitials: Your Initials (MRR)30: Incubation Temperature in CelsiusM: Media (T = Tryptic Soy Agar; N = Nutrient Agar; L = LB Agar; M = RDM; R = R2A; P = 1SP4)