Abstract
Theonella swinhoei is an arsenic hyper-accumulator sponge, harboring a multitude of associated bacteria. These bacteria reside in the mesohyl, the dense extracellular matrix of the sponge. Previous elemental analysis of separated cell fractions from the sponge had determined that arsenic is localized to the associated bacteria. Subsequently, sponge-associated arsenic-tolerant bacteria were isolated here and grouped into 15 operational taxonomic units (OTUs, 97 % similarity). Both culture-dependent and culture-independent work had revealed that T. swinhoei harbors a highly diverse bacterial community. It was thus hypothesized the acclimation of bacteria in the presence of a sponge skeleton, better mimicking its natural environment, would increase the yield of isolation of sponge-associated bacteria. Using seven modularly designed media, 380 bacteria isolates were grown and grouped into 22 OTUs. Inclusion of sponge skeleton in the growth medium promoted bacterial growth in all seven media, accounting for 20 of the 22 identified OTUs (the other two in a medium without skeleton). Diversity and richness indices were calculated for each treatment or combination of treatments with shared growth parameters. Integrating data inherent in the modularly designed media with the ecological indices led to the formation of new hypotheses regarding the aeration conditions and expected arsenic form in situ. Both aerobic and anoxic conditions are expected to occur in the sponge (temporally and/or spatially). Arsenate is expected to be the dominant (or even the only) arsenic form in the sponge.
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Acknowledgments
The authors would like to acknowledge the Interuniversity Institute, Eilat, and its staff for the ongoing support and use of their facilities. This work was partially supported by the Israel Science Foundation (grant number 957/14). Adi Lavy was funded by an Eshkol scholarship from the Israeli Ministry of Science, Technology and Space.
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Ray Keren and Adi Lavy contributed equally to this work.
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Fig. S1
The experimental design used to culture T. swinhoei-associated, arsenic-tolerant bacteria. A) Media used for culturing sponge-associated arsenic-tolerant bacteria from T. swinhoei. The difference between each media pair is expressed. B) Flow chart depicting the acclimation design leading to 21 growth conditions. Media abbreviation is composed of three parts: AsV or AsIII for arsenate and arsenite supplementation respectively. Aeration state is expresses as “O” for aerobic treatments and “an” for anoxic treatments. Media ending with “C” were supplemented with a carbohydrate mix while the others were supplemented with bicarbonate. S, N, and L stand for sulfide, nitrate, and light respectively follows (GIF 128 kb)
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Keren, R., Lavy, A. & Ilan, M. Increasing the Richness of Culturable Arsenic-Tolerant Bacteria from Theonella swinhoei by Addition of Sponge Skeleton to the Growth Medium. Microb Ecol 71, 873–886 (2016). https://doi.org/10.1007/s00248-015-0726-0
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DOI: https://doi.org/10.1007/s00248-015-0726-0