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Isolation and Identification of MDRO's in the Rio Grande River Poster
- 1. Abstract Abstract Abstract Abstract Abstract Abstract AbstractAbstract Hypothesis Acknowledgements Antibiotic resistant bacteriais 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