Using common marmoset (Callithrix jacchus) feces to isolate Escherichia coli

Presenter Information

grace walterFollow

Presenter Type

UNO Undergraduate Student

Major/Field of Study

Biology

Advisor Information

Professor

Location

CEC RM #201/205/209

Presentation Type

Poster

Poster Size

48”x38”

Start Date

22-3-2024 10:30 AM

End Date

22-3-2024 11:45 AM

Abstract

Using common marmoset (Callithrix jacchus) feces to isolate Escherichia coli

Kaitlyn Oswalda,*, Grace Waltera,*, Andrew K. Huanga, Mayowa Abioduna,b, Jonathan B. Claytona,b,c,d,e

*Authors contributed equally

aDepartment of Biology, University of Nebraska at Omaha, Omaha, NE, USA

bNebraska Food for Health Center, University of Nebraska-Lincoln, Lincoln, NE, USA

cDepartment of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE, USA

dDepartment of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE, USA

ePrimate Microbiome Project, University of Nebraska-Lincoln, Lincoln, NE, USA

The mammalian gut microbiome is home to a diverse range of bacteria which collectively function as an organ capable of influencing the host organism’s metabolic outcomes. A prominent member of the gut microbiome in mammals is Escherichia coli (E. coli). E. coli is a gram-negative bacterium that is commonly found in the lower intestine. To gain access to the bacteria, fecal samples were collected from Common Marmosets (Callithrix jacchus) housed at the Callitrichid Research Center (CRC) located at the University of Nebraska at Omaha. Following a six-day protocol, we recorded details regarding growth, viability, and presence/absence of contamination for all cultures. Choice of nutrient broth, selective agar, rpm speed of the shaking incubator, and glycerol concentrations were manipulated for this purpose. A specific and detailed procedure was followed on each day of the protocol, where 18-24 hours was allowed to pass between each phase. The samples were mixed in buffered peptone water facilitated by a shaking incubator at 37°C at 125 rpm. Four-quadrant streaking on MacConkey(MAC) agar was used to grow isolated colonies. After transferring the colonies onto Eosin Methylene Blue (EMB) agar with four-quadrant streaking, allowing them to grow into isolated colonies once more, the samples are transferred and stored in culture media in a 10-15% glycerol concentration. This has consistently yielded viable E. coli cultures. Expansion of the E. coli strain library is currently ongoing. As of March 8, 2024, we have been able to isolate E. colifrom seven marmosets. We have a total of 146 unique E. coli isolates stored at -80°C in the laboratory. Once it is complete, we plan to utilize the strain library for a series of endeavors. We plan to organize a study focused on examine antibiotic resistance profiles of the marmoset derived E. coli.

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Mar 22nd, 10:30 AM Mar 22nd, 11:45 AM

Using common marmoset (Callithrix jacchus) feces to isolate Escherichia coli

CEC RM #201/205/209

Using common marmoset (Callithrix jacchus) feces to isolate Escherichia coli

Kaitlyn Oswalda,*, Grace Waltera,*, Andrew K. Huanga, Mayowa Abioduna,b, Jonathan B. Claytona,b,c,d,e

*Authors contributed equally

aDepartment of Biology, University of Nebraska at Omaha, Omaha, NE, USA

bNebraska Food for Health Center, University of Nebraska-Lincoln, Lincoln, NE, USA

cDepartment of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE, USA

dDepartment of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE, USA

ePrimate Microbiome Project, University of Nebraska-Lincoln, Lincoln, NE, USA

The mammalian gut microbiome is home to a diverse range of bacteria which collectively function as an organ capable of influencing the host organism’s metabolic outcomes. A prominent member of the gut microbiome in mammals is Escherichia coli (E. coli). E. coli is a gram-negative bacterium that is commonly found in the lower intestine. To gain access to the bacteria, fecal samples were collected from Common Marmosets (Callithrix jacchus) housed at the Callitrichid Research Center (CRC) located at the University of Nebraska at Omaha. Following a six-day protocol, we recorded details regarding growth, viability, and presence/absence of contamination for all cultures. Choice of nutrient broth, selective agar, rpm speed of the shaking incubator, and glycerol concentrations were manipulated for this purpose. A specific and detailed procedure was followed on each day of the protocol, where 18-24 hours was allowed to pass between each phase. The samples were mixed in buffered peptone water facilitated by a shaking incubator at 37°C at 125 rpm. Four-quadrant streaking on MacConkey(MAC) agar was used to grow isolated colonies. After transferring the colonies onto Eosin Methylene Blue (EMB) agar with four-quadrant streaking, allowing them to grow into isolated colonies once more, the samples are transferred and stored in culture media in a 10-15% glycerol concentration. This has consistently yielded viable E. coli cultures. Expansion of the E. coli strain library is currently ongoing. As of March 8, 2024, we have been able to isolate E. colifrom seven marmosets. We have a total of 146 unique E. coli isolates stored at -80°C in the laboratory. Once it is complete, we plan to utilize the strain library for a series of endeavors. We plan to organize a study focused on examine antibiotic resistance profiles of the marmoset derived E. coli.