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

Presenter Information

Andrew HuangFollow

Presenter Type

UNO Undergraduate Student

Major/Field of Study

Biology

Other

Biology

Advisor Information

Jonathan B. Clayton

Location

MBSC Ballroom Poster # 805 - U

Presentation Type

Poster

Start Date

24-3-2023 10:30 AM

End Date

24-3-2023 11:45 AM

Abstract

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

Andrew K. Huang1, Emma Sorrell1, Paul Ayayee1, Aliyah Jabenis1,2, and Jonathan B. Clayton1,2,3,4

1Department of Biology, University of Nebraska at Omaha, Omaha, NE, USA

2Callitrichid Research Center, University of Nebraska at Omaha, Omaha, NE, USA

3Nebraska Food for Health Center, University of Nebraska-Lincoln, Lincoln, NE, USA

4Primate Microbiome Project, University of Nebraska-Lincoln, Lincoln, NE, USA

The mammalian gut microbiome is home to a conglomerate of diverse bacteria which collectively function as an organ capable of influencing the host organism’s metabolic outcomes. One of the most well-known members of the mammalian gut microbiome is Escherichia coli (E. coli), a gram-negative bacterium that typically resides in the lower intestine. Fecal samples collected from common marmosets housed at the Callitrichid Research Center (CRC) located at the University of Nebraska at Omaha were used to develop an effective and reproducible E. coli culture protocol. Factors such as choice of nutrient broth, selective agar, rpm speed of the shaking incubator, and glycerol concentration were manipulated for this purpose. We recorded details regarding growth, viability, and presence/absence of contamination for all cultures. Our results indicate that a protocol involving an initial growth period of 18-24 hours in buffered peptone water facilitated by a shaking incubator set at 37° C and 125 rpm, followed by rounds of four-quadrant streaking on MacConkey (MAC) agar until pure colonies are achieved, followed by four-quadrant streaking on Eosin Methylene Blue (EMB) agar, and storage of culture media in a 10-15% glycerol concentration, consistently yields viable E. coli cultures. Expansion of the E. coli strain library is currently ongoing. Once completed, we plan to utilize the strain library for a series of endeavors, including a study focused on examining antibiotic resistance profiles of the marmoset-derived E. coli.

Scheduling

10:45 a.m.-Noon, 1-2:15 p.m., 2:30 -3:45 p.m.

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Mar 24th, 10:30 AM Mar 24th, 11:45 AM

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

MBSC Ballroom Poster # 805 - U

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

Andrew K. Huang1, Emma Sorrell1, Paul Ayayee1, Aliyah Jabenis1,2, and Jonathan B. Clayton1,2,3,4

1Department of Biology, University of Nebraska at Omaha, Omaha, NE, USA

2Callitrichid Research Center, University of Nebraska at Omaha, Omaha, NE, USA

3Nebraska Food for Health Center, University of Nebraska-Lincoln, Lincoln, NE, USA

4Primate Microbiome Project, University of Nebraska-Lincoln, Lincoln, NE, USA

The mammalian gut microbiome is home to a conglomerate of diverse bacteria which collectively function as an organ capable of influencing the host organism’s metabolic outcomes. One of the most well-known members of the mammalian gut microbiome is Escherichia coli (E. coli), a gram-negative bacterium that typically resides in the lower intestine. Fecal samples collected from common marmosets housed at the Callitrichid Research Center (CRC) located at the University of Nebraska at Omaha were used to develop an effective and reproducible E. coli culture protocol. Factors such as choice of nutrient broth, selective agar, rpm speed of the shaking incubator, and glycerol concentration were manipulated for this purpose. We recorded details regarding growth, viability, and presence/absence of contamination for all cultures. Our results indicate that a protocol involving an initial growth period of 18-24 hours in buffered peptone water facilitated by a shaking incubator set at 37° C and 125 rpm, followed by rounds of four-quadrant streaking on MacConkey (MAC) agar until pure colonies are achieved, followed by four-quadrant streaking on Eosin Methylene Blue (EMB) agar, and storage of culture media in a 10-15% glycerol concentration, consistently yields viable E. coli cultures. Expansion of the E. coli strain library is currently ongoing. Once completed, we plan to utilize the strain library for a series of endeavors, including a study focused on examining antibiotic resistance profiles of the marmoset-derived E. coli.