BUILDing Scholar Oral Prize Winner

1. Abstract
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Hypothesis
Acknowledgements
Antibiotic resistant bacteria is becoming a severe and imminent threat to
public health worldwide. Antibiotics are commonly used to treat a widespread
range of bacterial infections. Their accessibility and improper use allows
bacteria to build resistance over time to antibiotics. The Rio Grande River is
1,885 miles long, crossing through three states and even making up part of
the U.S-Mexico border. This makes it an important resource, and if antibiotic
resistant bacteria were to be found, it would imply that a large issue is at
hand. To determine if antibiotic resistant bacteria was present, water samples
from two sites in the Rio Grande were filtered and isolated to test for
antibiotic resistance. After incubating the filters, they were streaked for
isolation and incubated once again. A day later, the bacteria was gram stained
for identification purposes, which enables us to know what antibiotic plate to
use. Once inoculated and incubated, the panels have reagents added to them
which completes the process. The panels were then analyzed in the
MicroScan autoSCAN 4 to display the bacteria’s genus, species, minimum
inhibitory concentration (MIC), and resistances. This bacterial identification
system has a 99.99% accuracy rate. Subsequently, the results were compiled
and analysis showed that the Rio Grande River has antibiotic resistant
bacteria; nineteen of the thirty-four results had antibiotic resistance to four
classes of antibiotics, including carbapenems.
The Rio Grande River is the fifth longest river in the United States, and is part
of the United States-Mexico border. It has agricultural, recreational, and
domestic purposes, which is important to states it runs through, such as
Texas, Colorado, and New Mexico. Traces of pathogenic bacteria have been
found within the Rio Grande River in the past, some of the commonly found
bacteria is E.coli, a naturally present flora in the digestive system. When E.coli
is in water sources it can be a sign of fecal contamination. The problem with
bacteria like E.coli is that if it possesses antibiotic resistant properties, those
resistances can be shared with other bacteria. This could render commonly
used clinical antibiotics insufficient in stopping infections. The objective of this
study is to isolate and identify antibiotic resistant bacteria using the
MicroScan autoSCAN-4 system.
Antibiotic resistant bacteria will be found in the Rio Grande River.
We would like to acknowledge our mentors: Dr. Alvarez, Joe Mendoza, Jose Gomez and
Daniella Sahagun for helping us and showing us the direction to go, as well as the rest of the
laboratory members who all aided us along the way. Research reported in this poster was
supported by the National Institute of General Medical Sciences of the National Institutes of
Health under linked Award Numbers RL5GM118969, TL4GM118971, and UL1GM118970. The
content is solely the responsibility of the authors and does not necessarily represent the
official views of the National Institutes of Health.
Antibiotic resistant bacteria was found in the Rio Grande River. A high
number of the antibiotic resistance bacteria found was resistant to a
minimum of 5 antibiotics, with E.coli being one of the bacteria encountered.
The presence of E.coli could signal that there is fecal contamination in the
river, which would prompt to further research about the river. A high number
of possible ESBL’s were also detected, which could prompt another research
opportunity relating to ESBL concentration in the Rio Grande river. All
possible Extended Spectrum Beta-Lactamase results have been sent to UTEP
for further investigation and confirmation.
Isolation and Identification of Antibiotic Resistant Bacteria In The Rio Grande River
Ray Trujillo, Kaylee Wersant, Joe Mendoza, Dr. Maria Alvarez
Abstract
Introduction
Materials and Methods
Results
Results
Conclusion
El Paso Independent School District, El Paso Community College
Step 2 – Filter water through 0.45 μm Millipore filter in sets of 2ml, 5ml, and 10ml.
Step 4 – Gram Staining.
Using the filtration system, we filtered water in 2ml, 5ml, and 10ml sets
onto media. The overall number of filters for all sites after incubation
was 36 filters.
Gram staining is a vital part of the process, as the results decide
what panel (positive or negative) to inoculate the bacteria in.
Step 3 – Streaking for isolation.
To streak for isolation, a single isolated colony is
required to streak. The colony is then streaked
onto its corresponding media.
Step 5 – Inoculating, Paneling, and Results.
A colony was then inoculated and incubated. After the
incubation period (24hrs), the panels have reagents
added to them and they were prepped for the
microScan AutoSCAN 4.
Materials
Filtration System Inoculation Wands Empty RENOK Plate
Methods
Positive Antibiotic
Combo Panel
MicroScan AutoSCAN 4 w/
Computer System
E.coli 99.99% MIC Final
Amikacin >32 R
Amox/ K Clav >16/ 8 R
Amp/ Sulbactam>16/ 8 R
Ampicillin >16 R
Aztreonam >16 EBL?
Cefazolin >16 R
Cefepime >16 R
Cefotaxime >32 EBL?
Cefotetan >32 R
Cefoxitin >16 R
Ceftazidime >16 EBL?
Cefrtiaxone >32 EBL?
Cefuroxime >16 R
Cephalothin >16
Chloramphenicol>16 R
Ciprofloxacin >2 R
ESBL-a Scrn >4 EBL?
ESBL-b Scrn >1 EBL?
Gatifloxacin >4 R
Gentamicin >8 R
Imipenem >8 R
Levofloxacin >4 R
Meropenem >8 R
Moxifloxacin >4 R
Nitrofurantoin >64
Norfloxacin >8
Pip/ Tazo >64 R
Piperacillin >64 R
Tetracycline >8 R
Ticar/ K Clav >64 R
Tobramycin >8 R
K.Pneumoniae - 99.99% MIC Flags
Amikacin >32 R
Amox/ K Clav >16/ 8 R
Amp/ Sulbactam >16/ 8 R
Ampicillin >16 R
Aztreonam >16 EBL?
Cefazolin >16 R
Cefepime >16 R
Cefotaxime >32 EBL?
Cefotetan >32 R
Cefoxitin >16 R
Ceftazidime >16 EBL?
Ceftriaxone >32 EBL?
Cefuroxime >16 R
Cephalothin >16
Chloramphenicol >16 R
Ciprofloxacin >2 R
ESBL-a Scrn >4 EBL?
ESBL-b Scrn >1 EBL?
Gatifloxacin >4 R
Gentamicin >8 R
Imipenem >8 R
Levofloxacin >4 R
Meropenem >8 R
Moxifloxacin >4 R
Nitrofurantoin
Norfloxacin
Pip/ Tazo >64 R
Piperacillin >64 R
Tetracycline >8 R
Ticar/ K Clav >64 R
Tombramycin >8 R
S.xyosus - (99.55%) MIC Final
Amox/ K Clav <=4/ 2 R
Amp/ Sulbactam <=8/ 4 R
Ampicillin <=0.25 R
Azithromycin <=2 S
Cefazolin <=8 R
Cefepime <=8 R
Cefotaxime <=8 R
Ceftriaxone <=8 R
Cephalothin <=8 R
Chloramphenicol <=8 S
Ciprofloxacin <=1 S
Clindamycin 2 I
Eryhromycin <=0.5 S
Gatifloxacin <=2 S
Gentamicin <=4 S
Imipenem <=4 R
Levofloxacin <=2 S
Linezolid
Moxifloxacin <=2 S
Norfloxacin
Nitrofurantoin
Ofloxacin <=2 S
Oxacillin 1 R
Penicillin 0.12 R
Pip/ Tazo
Rifampin <=1 S
Synercid <=1 S
Tetracycline <=4 S
Trimeth/ Sulfa <=2/ 38 S
Step 1 – Gathering water samples from the Rio Grande River
Gather water samples from Anapra and Corchense, which can
be located on the map to the bottom left corner.
Bacterial
Identification No. of Isolates
Probability of
Correct
Identification
No. of Isolates with
resistance to more than 4
4 classes of antibiotics
A.xylosoxidants 1 99.99% 0
Aer hydro cplx 4 82.50%-99.99% 1
Cedecea davisae 2 99.99% 2
E.coli 5 99.99% 3
Gemella spp. 1 96.71% 0
K.oxytoca 2 99.18% 2
K.pneumoniae 10 93.94%-99.99% 9
Leminorella 1 86.26% 0
Micro/Rel spp. 2 81.43%-91.40% 0
P.aeruginosa 1 86.30% 0
P.stuartii 1 87.01% 0
S.auricularis 1 99.09% 0
S.xylosus 2 99.55% 0
V.cholerae 1 47.49% 0
V.fluvalis 1 85.43% 0
V.parahaemolyt 1 93.46% 1
Bacterial
Identification No. of Isolates Possible ESBL’s
A.xylosoxidants 1 0
Aer hydro cplx 4 0
Cedecea davisae 2 0
E.coli 5 3
Gemella spp. 1 0
K.oxytoca 2 2
K.pneumoniae 10 9
Leminorella 1 0
Micro/Rel spp. 2 0
P.aeruginosa 1 0
P.stuartii 1 0
S.auricularis 1 0
S.xylosus 2 0
V.cholerae 1 0
V.fluvalis 1 0
V.parahaemolyt 1 0
Total 36 14