Using common marmoset (Callithrix Jacchus) feces to isolate and characterize Acinetobacter
Presenter Type
UNO Undergraduate Student
Major/Field of Study
Physics
Advisor Information
jclayton@unomaha.edu (Jonathan B. Clayton)
Location
CEC RM #201/205/209
Presentation Type
Poster
Poster Size
24x36
Start Date
22-3-2024 9:00 AM
End Date
22-3-2024 10:15 AM
Abstract
Andrew Kiyoji Huang1, Mayowa Abiodun1,2, Jordan B. Hernandez1,2,3, Kurt Piepenbrink2,4, and Jonathan B. Clayton1,2,4,5,6
1Department of Biology, University of Nebraska at Omaha, Omaha, NE 68182, United States
2Nebraska Food for Health Center, University of Nebraska-Lincoln, Lincoln, NE 68588, United States
3Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE 68198, United States
4Department of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE 68588, United States
5Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, United States
6Primate Microbiome Project, University of Nebraska-Lincoln, Lincoln, NE 68588, United States
Acinetobacter is a gram-negative non-motile microbe infamous for the danger it poses in the clinical setting. Although it has low virulence, Acinetobacter’s wide range of drug resistances has proven deadly for immunocompromised patients. Recently, one of our lab members detected the Acinetobacter genus within the gut microbiome of several common marmosets (Callithrix Jacchus) housed at the University of Nebraska at Omaha using a tool called SPIECE-EASI. In collaboration with the Piepenbrink Lab at the University of Nebraska-Lincoln, we attempted to isolate Acinetobacter from fecal samples collected from the marmosets. We obtained isolates using a series of selective media which included an initial growth period in MacConkey (MAC) Broth and one to two rounds of streaking on CHROMagar AcinetobacterTM (CHROM) using four-quadrant streaking. A centrifuge was utilized to separate the culture into a pellet which was resuspended in MAC broth and finally frozen at -80 °C in a 1:1 mixture of 80% glycerol solution and the MAC-culture mixture. Obtained isolates were then sent to the Piepenbrink Lab, where they performed a soft agar swimming test to determine the motility of the suspect isolate. The presentation of our isolate on the CHROMagar, MAC Agar, and the soft agar motility test suggests the isolate may be a species of Acinetobacter. On CHROMagar, colonies of the suspect isolate demonstrated a light blue coloration. According to the manufacturer of CHROM, Acinetobacter baumannii presents red-colored colonies on CHROM while a blue presentation indicates another gram-negative bacterium; this still leaves the possibility for another strain of Acinetobacter. On MAC agar, the isolate showed that it fermented lactose very slowly, if not at all; this behavior is consistent among Acinetobacter. The motility test, on the other hand, was inconsistent. The suspect isolate demonstrated high motility with a spread akin to a featureless mat, while the known Acinetobacter isolate showed inconsistent motility with one of four plates featuring the featureless mat while the others showed little to no spread. DNA sequencing will be necessary to confirm the identity of the isolate. Future applications of this protocol will mix and thaw fecal samples to increase the likelihood of culturing Acinetobacter. The presence of Acinetobacter in the marmoset gut microbiome may serve as a tool for measuring the effects of antibiotic-induced dysbiosis in a marmoset model. Outbreaks of Acinetobacter and consequential infections may indicate a weakened immune system as a product of depression stemming from antibiotic-induced gut dysbiosis.
Using common marmoset (Callithrix Jacchus) feces to isolate and characterize Acinetobacter
CEC RM #201/205/209
Andrew Kiyoji Huang1, Mayowa Abiodun1,2, Jordan B. Hernandez1,2,3, Kurt Piepenbrink2,4, and Jonathan B. Clayton1,2,4,5,6
1Department of Biology, University of Nebraska at Omaha, Omaha, NE 68182, United States
2Nebraska Food for Health Center, University of Nebraska-Lincoln, Lincoln, NE 68588, United States
3Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE 68198, United States
4Department of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE 68588, United States
5Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, United States
6Primate Microbiome Project, University of Nebraska-Lincoln, Lincoln, NE 68588, United States
Acinetobacter is a gram-negative non-motile microbe infamous for the danger it poses in the clinical setting. Although it has low virulence, Acinetobacter’s wide range of drug resistances has proven deadly for immunocompromised patients. Recently, one of our lab members detected the Acinetobacter genus within the gut microbiome of several common marmosets (Callithrix Jacchus) housed at the University of Nebraska at Omaha using a tool called SPIECE-EASI. In collaboration with the Piepenbrink Lab at the University of Nebraska-Lincoln, we attempted to isolate Acinetobacter from fecal samples collected from the marmosets. We obtained isolates using a series of selective media which included an initial growth period in MacConkey (MAC) Broth and one to two rounds of streaking on CHROMagar AcinetobacterTM (CHROM) using four-quadrant streaking. A centrifuge was utilized to separate the culture into a pellet which was resuspended in MAC broth and finally frozen at -80 °C in a 1:1 mixture of 80% glycerol solution and the MAC-culture mixture. Obtained isolates were then sent to the Piepenbrink Lab, where they performed a soft agar swimming test to determine the motility of the suspect isolate. The presentation of our isolate on the CHROMagar, MAC Agar, and the soft agar motility test suggests the isolate may be a species of Acinetobacter. On CHROMagar, colonies of the suspect isolate demonstrated a light blue coloration. According to the manufacturer of CHROM, Acinetobacter baumannii presents red-colored colonies on CHROM while a blue presentation indicates another gram-negative bacterium; this still leaves the possibility for another strain of Acinetobacter. On MAC agar, the isolate showed that it fermented lactose very slowly, if not at all; this behavior is consistent among Acinetobacter. The motility test, on the other hand, was inconsistent. The suspect isolate demonstrated high motility with a spread akin to a featureless mat, while the known Acinetobacter isolate showed inconsistent motility with one of four plates featuring the featureless mat while the others showed little to no spread. DNA sequencing will be necessary to confirm the identity of the isolate. Future applications of this protocol will mix and thaw fecal samples to increase the likelihood of culturing Acinetobacter. The presence of Acinetobacter in the marmoset gut microbiome may serve as a tool for measuring the effects of antibiotic-induced dysbiosis in a marmoset model. Outbreaks of Acinetobacter and consequential infections may indicate a weakened immune system as a product of depression stemming from antibiotic-induced gut dysbiosis.