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Accessing Antibiotic Resistance in Pristine vs Anthropogenic Impacted Environments Using Metagenomics

Undergraduate #112
Discipline: Biological Sciences
Subcategory: Microbiology/Immunology/Virology

Ricky Padilla-Del Valle - University of Puerto Rico at Mayaguez
Co-Author(s): Carlos Rios-Velazquez, University of Puerto Rico at Mayaguez



The outbreak of infectious diseases with Antibiotic Resistance (AR) has threat the first line of the medical defense in the world. During years, classic microbiology tried to resolve this problem by producing new antibiotics but its continuous overuse only increased the frequency of AR pathogenic infections. In other hand, functional metagenomic analysis have reveal that world’s ecosystems have a diverse collection of AR genes. The purpose of our research was to generate metagenomic libraries (ML) from two different types of conditions: from a pristine environment and a human anthropogenic impacted ecosystem (HAIE). Then, explore for AR genes against ampicillin, kanamycin and tetracycline. Two ML from compost, a vegetable waste and a human biosolid, were generated (representing HAIE) and two ML from soils of Isla de Mona, PR were also generated (representing pristine environment). The high molecular weight (40kbp) DNA of each one was end-repaired, electro eluted, and ligated into the fosmid vector pCCFOS1, then, transduced to Escherichia coli Epi300-T1R using T1 bacteriophage. The Minimal Inhibitory Concentration (MIC) of ampicillin, kanamycin and tetracycline were determined. The AR clones from the libraries were isolated by selection on culture media supplemented with 1X-10X MIC of each antibiotic separately. While many clones in the ML of composts showed AR to 1X-10X MIC of ampicillin, no AR to ampicillin was detected in the clones from the pristine environment. These results represent that HAIE are potential reservoirs of β-lactams AR in contrast to the pristine environment. Further analysis will confirm AR against the other two antibiotics.

Funder Acknowledgement(s): Puerto Rico Louis Stroke Alliance for Minority Participation (Grant number: 1400868); RISE to Best - University of Puerto Rico (NIH: R25GM088023).

Faculty Advisor: Carlos Rios-Velazquez, carlos.rios5@upr.edu

Role: All the research project was done by the undergraduate student.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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