This document summarizes the isolation of bacteria from contaminated soils in Puerto Rico. Soil samples were collected from four locations across Puerto Rico and tested to isolate bacteria. Three distinct bacteria strains (CLRV30TSA01A, CLRV30M01B, and JCR30TSA01A) were isolated based on gram staining, morphology, PCR/electrophoresis testing, and antibiotic production profiles. These three isolates were then tested for antibiotic resistance against several common antibiotics. Two additional isolates (S15UPRC-RISESCDM30T01B and S15UPRC-RISEPR30M01) were also tested for antibiotic resistance.

1. ISOLATING BACTERIA FROM TROPICAL
CONTAMINATED SOILS IN PUERTO RICO
Sara Diaz
Patricia Rodriguez
Camille Ruiz
Justin Cotto

2. SAMPLING LOCATIONS
Sample Coordinates Environmental description Location Date/Time
Ruiz C
- #1
18°6’55.1”N;
66°9’36.2”W
Moist (underneath grass), 5 cm;
72°F
Cayey, PR January/28/15
7:30 p.m.
Cotto J
- #2
18.15º N
66.18º W
Moist environment (above
grass), 76ºF
Cidra, PR January/29/15
8:00 a.m.

3. SAMPLING LOCATIONS
Sample Coordinates Environmental description Location Date/Time
Diaz S
-#1
18°15'15.7"N
66°06'06.9"W
Moist environment
80°F
Aguas
Buenas, P.R.
4:26 p.m.
Rodríguez P
-#2
18°15'15.7"N
66°06'06.9"W
Moist environment
80°F
Aguas
Buenas, P.R.
4:26 p.m.

4. Process CLRV30TSA01A CLRV30M01B JCR30TSA01A
Grand Stain Negative Positive Positive
Bacterial Morphology Coccus Bacillus Bacillus
PCR/Electrophoresis #1 Negative Negative Negative
PCR/Electrophoresis #2 Positive Negative -
Antibiotic Production Negative Negative Negative

5. Process S15UPRC-RISESCDM30T01B S15UPRC-RISEPR30M01
Gram Stain Positive Positive
Bacterial Morphology Bacillus Bacillus
PCR/Electrophoresis #1 Negative Negative
PCR/Electrophoresis #2 Negative Negative
Antibiotic Production Negative Negative

6. ANTIBIOTIC RESISTANCE TEST
RESULTS
Antibiotic CLRV30TSA01A CLRV30M01B JCR30TSA01A
Kanamycin 2.0 cm 1.0 cm 2.0 cm
Penicillin 0.0 cm 0.0 cm -
Tetracycline 1.6 cm 2.4 cm 2.2 cm
Gentamicin 1.9 cm 0.9 cm -
Chloramphenicol 0.0 cm 0.0 cm 2.5 cm
Bacitracin 0.0 cm 0.0 cm 1.5 cm
Vancomycin 0.0 cm 0.0 cm -
Streptomycin 1.6 cm 1.0 cm 1.9 cm

7. ANTIBIOTIC RESISTANCE TEST
RESULTS
Antibiotic S15UPRC-RISESCDM30T01B
Streptomycin 2.2 cm
Chloramphenicol 2.3 cm
Erythromycin 1.0 cm
Kanamycin 1.6 cm
Gentamicin 0.6 cm
Antibiotic S15UPRC-RISEPR30M01
Rifampin 0.13 cm
Kanamycin 0.16 cm
Vancomycin 0.17cm
Streptomycin 0.30 cm
Tetracyclin 0.14 cm