Background
Methods and Materials
Objective
Results
Conclusion
Acknowledgements
References
Use of an Automated Bacterial Identification System & PCR Analysis to test for the Presence of E. Coli 0157:H7 in the Rio Grande River
K. Reynozo2, N. Holguin2, M. Zeschitz2, J. Mendoza1, M. E. Alvarez1, C. Chianelli¹
El Paso Community College,1El Paso ,Texas, Transmountain Early College High School, El Paso , Texas
The aim of this project was to determine if there is contamination of the Rio Grande River by E. coli 0157:H7 strain. E. coli is a symbiotic and a gram negative bacteria that is commonly found in the intestines of humans and other animals. E. Coli 0157:H7 is a bacteria that can cause life threatening symptoms such as bloody diarrhea, severe anemia, kidney failure, and urinary tract infections (UTI). Many people become infected with E. coli by eating or drinking contaminated food that has come in contact with animal feces. The Siemens Micro Scan was used to detect E. Coli. The Norgen Biotek E. Coli 0157:H7 PCR detection kit was then used to test for the presence of E. Coli 0157:H7. None of the isolates tested positive for the E. Coli 0157:H7 strain.
E. coli is a gram negative bacteria that is commonly found in the intestines of humans and other animals.
There are some E. coli like the 0157:H7 strain that can cause life threatening symptoms such as kidney failure, bloody diarrhea, urinary tract infections, and severe anemia
Many people become infected with E. coli by eating or drinking contaminated food with feces.
Since the river is near dairy farms, the waste of these farms runoff into the river which causes it to get contaminated with E. Coli.
People are then exposed with this pathogenic bacteria(s) through the products that are irrigated with the water of the river.
To test the water of the river to determine the amount of E. Coli that resides in the rivers‘ water.
We would like to acknowledge:
We still have several isolates that could be positive for future study, none of the E. Coli was positive for the 0157:H7 from the Courchesne Bridge (site 2).
Our results came out negative for the 0157:H7 E.Coli strain
Extraction of Samples: taking water samples from the Rio Grande River at the Courschesne Bridge (site 2)
Filtering: filter membrane (0.45um) , automated pipet, Bunsen burner, striker, tweezers, distilled water, sample from site 2 (Courschesne Bridge), filtration unit
Isolation: Bunsen burner, m.Endo , TSA, mTEC, inoculation wands , Petri dishes, & light microscope.
Inoculation: Micro Scan Neg/Urine combo panel type 35, inoculation kit, mineral oil, the various chemicals that correspond to each well, Siemens Micro Scan auto SCAN 4
PCR: Norgen Biotek E. Coli 0157:H7 PCR detection kit, Fisher Scientific Vortex Mixer, Eppendorf Centrifuge5415 D, horizontal electrophoresis, Bio-Rad power supply, automated pipets, DNA stain
Other units Used: flame hood, incubator, autoclave, freezer
Organism | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli | E. Coli |
Isolate Number | 3 | 18 | 15 | 36 | 41 | 32 | 35 | 40 | 38 | 43 | 31 | 39 | 37 | 29 | 34 | 44 | 42 | 33 | 45 |
% Probability ID | 99.99% | 99.99% | 99.99% | 99.99% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% | 96.51% |
The other bacteria found was Shigella at 3.49%.