Histology and Cell Culture of Octocoral, Antillogorgia americana, to Enhance their Potential for Genomics and Conservation
Defense Date
8-9-2024
Document Type
Thesis
Degree Type
Master of Science
Degree Name
Biological Sciences
First Advisor
Dr. Jose V. Lopez
Second Advisor
Dr. Geeta Ravindran
Third Advisor
Dr. Timothy Swain
Keywords
Cnidarians, cell culture, histology, microbial community, Next Generation Sequencing (NGS)
Abstract
Cnidarians have been used as model organisms in developmental and molecular biology for decades and are an excellent system to study cellular biology of organisms through cell culture. However, establishing cnidarian cell cultures has been a major challenge due to the difficulty in determining the most beneficial media, antibiotics, and dissociation methods that yield viable cell types, stave off contamination, and maximize cell growth. Few studies address interactions of multiple cells or how antibiotics affect the cnidarian microbiome in cell culture. Additionally, there are few studies characterizing the dominant taxa in cnidarian cultures. Using an octocoral species common to the Florida Reef Tract, Antillogorgia americana (Gmelin,1791), I ran cnidarian primary cell cultures for seven days to observe interactions with natural or contaminating microbes. Specifically, various cell culture methods were investigated and manipulated to enhance cnidarian cell growth. A. americana cells appear to be present up to day 7 of culturing via PCR assays. I characterized dominant microbial taxa, as well as other microbes found within cnidarian microbiome using 16S rRNA High Throughput Sequencing (HTS). A. Americana samples were collected at local reefs near Dania Beach, FL from September 2023 to May 2024. A total of 24 culture samples and 10 tissue samples were processed for 16S rRNA HTS, generating an average of 219,603 reads per sample. The dominant phyla found in tissue samples were Proteobacteria, Bacteriodota, Firmicutes, Desulfobacterota, and Verrucomicrobiota whereas, the culture samples included Proteobacteria, Bacteriodota, Planctomycota, and Desulfobacterota. Microbial beta diversity analyses showed noticeable changes over the culture period. A shift in bacterial abundances between A.americana culture and tissue samples was observed at the family level. This study provides evidence of a baseline relationship between the cnidarian microbiome and its potential impact on establishing successful cnidarian cell cultures.
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NSUWorks Citation
Rachel H. Bacaner. 2024. Histology and Cell Culture of Octocoral, Antillogorgia americana, to Enhance their Potential for Genomics and Conservation. Master's thesis. Nova Southeastern University. Retrieved from NSUWorks, . (204)
https://nsuworks.nova.edu/hcas_etd_all/204.