Microbial Diversity Associated with the Oral and Fecal Samples of White-Nose Syndrome (WNS) Infected Tricolored Bats

Graduate #10
Discipline: Biological Sciences
Subcategory: Genetics
Session: 1
Room: Virginia B

Kristina Burns - Alabama A&M University
Co-Author(s): William Stone, Department of Biological & Environmental Sciences, Alabama A&M University, Normal, AL, USA; Jeanette Jones, Department of Biological & Environmental Sciences, Alabama A&M University, Normal, AL, USA; David Laurencio, Museum of Natural History, Auburn University, AL, USA; Govind C. Sharma, Department of Biological & Environmental Sciences, Alabama A&M University, Normal, AL, USA; Venkateswara R. Sripathi, Department of Biological & Environmental Sciences, Alabama A&M University, Normal, AL, USA

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White-nose syndrome (WNS) has spread rapidly across the eastern United States and Canada since it was first documented in New York over a decade ago. The causative organism for WNS in bats is Pseudogymnoascus destructans. Pd is accountable for the 6.7 million deaths of bats in the United States. Molecular mechanisms associated with White-nose syndrome and the resultant shift in the microbial diversity are underexplored. Species identification was achieved by using both field-based methods and molecular techniques. This study included one bat species (Tricolored, Perimyotis subflavus); one condition (WNS-infected); one population (males); two sample types (oral and fecal swabs) with eighteen replicates (R1 – R18). In total, 36 microbiome samples (1 x 1 x 1 x 2 x 18) were selected for microbial DNA extraction (ZymoBIOMICS). The best practices in preserving bat microbiome samples for molecular analyses were tested by comparing DNA isolated from two different sample types (oral and fecal) using two different methods (spectrophotometer-based and quantitative PCR) and three different preservation media (ethanol, NaCl-saturated dimethyl sulfoxide (DMSO), and silica desiccant) and suggested that the DNA isolated from fecal samples and silica yielded more DNA. Microbial diversity was assessed by sequencing single-end libraries of amplified 16S rDNA genes from respective microbiomes and analyzed using Qiime 2. The reads were clustered into operational taxonomic units (OTUs), and phylogenetic trees were constructed with closely related bacterial species to compare microbiomes. Preliminary results indicated that the majority of OTUs identified belonged to the phylum Spirochaetes and Leptospira being the dominant genus. Further, bats being a mammalian system, the knowledge generated here can be applied to humans.

Funder Acknowledgement(s): Funding was provided by Title III Strengthening Grants Programs.

Faculty Advisor: Venkateswara R. Sripathi, v.sripathi@aamu.edu

Role: Field collection in local caves was completed under supervision by Dr. William Stone. All extractions and data analysis was performed by myself with multiple reviews by my advisor, Dr. Sripathi.