Primary co-culture as a complementary approach to explore the diversity of bacterial associations in marine invertebrates: the example of Nautilus macromphalus (Cephalopoda: Nautiloidea)
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The recent application of molecular tools to address associations between bacteria and marine invertebrates has provided access to an immense diversity of unidentified microbes resistant to cultivation. However, the role of bacteria as partners in animal physiology remains unclear and in most cases difficult to investigate in the absence of adequate condition of cell growth and proliferation. In this work, we studied the reservoir of microbes associated with the excretory organs of Nautilus macromphalus as a model. Using the bacterial 16S RNA gene as a marker, we compared three complementary approaches for bacterial detection: bacterial DNA extraction from N. macromphalus tissues (“molecular approach”), strain isolation to provide a bacterial culture collection (“microbiological approach”) and finally, maintenance of N. macromphalus excretory organ cells with associated bacteria (“cellular approach”). Our results stress the potential of the “cellular approach” as a promising new tool as it promotes the detection of as yet uncultured β-proteobacteria and spirochaetes associated with N. macromphalus, and serves as a foundation for future studies describing potential roles that these bacteria may play in Nautilus.
KeywordsMarine Agar Pseudomonadales Cellular Approach Microbiological Approach Alteromonadales
We thank P. Joannot, S. Loueckote, A. Gerbault and C. Goiran for their help in providing N. macromphalus specimens from New-Caledonia. The help of C. Courties for flow cytometry analysis was highly appreciated. We thank A. Andouche and F. Ponton for helpful comments on this manuscript. This work was supported by a Bonus Quality Research grant from the Muséum National d’Histoire Naturelle (PARIS). The French Ministry for National Education and Research is also acknowledged for providing M. Pernice with a Ph.D. grant. The experiments complied with the current French laws.
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