Date of Award
Summer 7-31-2023
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Biology
First Advisor
Anuradha Ghosh
Second Advisor
Andrew George
Third Advisor
Jennifer Harris
Keywords
Metagenomic gray bats bacteria dna sequencing
Abstract
USING A CULTURABLE APPROACH AND METAGENOMIC ANALYSIS TO UNCOVER GUT BACTERIAL DIVERSITY OF GRAY BATS IN KANSAS
An Abstract of the Thesis by
Bobbi Monroe
Humans have historically had an ambivalent relationship with bats. In one hand, bats perform an important service to humans by reducing populations of many insect pests. On the other hand, they act as reservoirs of disease as highlighted by the recent Coronavirus pandemic. In the United States, many bat populations have been threatened by white nose syndrome caused by the fungus Pseudogymnoascus destructans. This study was designed to characterize the bacterial diversity associated with the gray bat (Myotis grisescens) in Southeast Kansas. In addition, this study aimed to shed light on the presence of bacterial pathogens, if any, among this bat population. A total of 32 guano samples were collected and bacterial isolates with different colony morphology were recovered on tryptic soy agar media after enrichment. The majority (21/32, 65%) of isolates were Gram positive. All isolates were tested for growth on selective and differential media. Sugar fermentation profiles showed that 78% (25 of all isolates) fermented all four sugars, 9% (3/32) fermented three sugars, another 9% (3/32) fermented two sugars, and one isolate (1/32) 3% fermented only one sugar. Urea was hydrolyzed by seven (21%) isolates while one isolate (3%) was positive for indole production. After bacterial isolation results were shortlisted to removed possible repeats, additional tests yielded that 52% (13/25) were one step nitrate reducers, 68% (17/25) were MR positive while 32% (8/25) were VP positive, 52% (13/25) were oxidase positive and only 0.04% (1/25) was positive for phenyl alanine. All isolates were tested for their susceptibility to multiple antibiotics. In addition to biochemical characterization, the isolates were identified using molecular techniques. Pooled samples were sequenced using an Illumina mini sequencer. A total of 2,909,555 reads were completed. The most common Gram positive genera being Bacillus (17.01%) and Lysinibacillus (19.93%) while the most common Gram negative genera were Serratia (26.36%) and Achromobacter (20.17%). Male and Female samples were sequenced using the same Illumina mini sequencer. For males a total of 5,408,935 reads were completed and for females 9,645,398 reads were completed. The most common species in male Gray Bats were Bacillus pakistanensis (17%) followed by Enterococcus faecalis (7.17%) and Jeotgalicoccus pinnipedialis (2.70%). The most common species in female Gray Bats were Romboustia clostridium (11%), followed by Enterrococcus faecalis (7.9%), Bacillus pakinstanensis (3.90%). The identification of both known and novel bacteria in bats is important for prevention of possible disease spread and in preserving the bat species further.
Recommended Citation
Monroe, Bobbi, "USING A CULTURABLE APPROACH AND METAGENOMIC ANALYSIS TO UNCOVER GUT BACTERIAL DIVERSITY OF GRAY BATS IN KANSAS" (2023). Electronic Theses & Dissertations. 461.
https://digitalcommons.pittstate.edu/etd/461
Comments
Revision draft