Dr. Joel Chandlee
Cell and Molecular Biology
Tedania ignis; Bermuda; Clone library; High Microbial Abundance
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Sponges are filter-feeding organisms that contain a dense and diverse microbial community. These bacteria and archaea can comprise about 40% of the sponges’ total biomass often exceeding the microbial biomass of seawater by two to three orders of magnitude. The presence of bacteria during the reproductive stages of the sponge is an indicator of symbiosis. This study used culture-independent techniques to investigate the microbial community of Tedania ignis; an abundant marine sponge in the inshore coral reef environments around Bermuda. Sponge and water samples were collected from Ferry Reach, Helena’s Bay and Bailey’s Bay in Bermuda. Epifluorescent microscopy was used to quantify the microbial abundance from within the sponge tissue and compare it to that of the surrounding water. T. ignis was categorized as an high microbial abundant sponge. Two extraction methods were used to isolate bacterial and archaeal DNA for clone library analysis of the microbial community composition. The Mo Bio UltraClean Soil DNA Isolation Kit was more successful at amplifying bacterial DNA while the CTAB method had higher DNA yields and purity but residual phenol may have led to PCR inhibition. Clone sequence analysis showed inconclusive results when the 27F primer sequence did not align well with the sample sequences. T. ignis was determined to harbor Synechococcus sp., Actinosynnema mirum, Collimonas fungivorans, Rhodococcus opacus and Candidatus Puniceispirillum marinum based on the BLAST sequence alignment. Water samples contained Actinosynnema mirum, and Paenibacillus graminis based on the BLAST sequence alignment.