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  1. Mesophotic coral reef ecosystems (MCEs) are characterized by gradients in irradiance, temperature and trophic resources. As depth increases on Caribbean mesophotic reefs, particulate organic matter increases while dissolved organic matter decreases, and the increase in particulate organic matter is directly related to the increase in sponge abundances and growth rates on MCEs. To further understand the trophic ecology of sponges, changes in microbiome composition and function, stable isotopic composition and proximate biochemical composition of 4 Caribbean reef sponges ( Amphimedon compressa , Agelas tubulata , Plakortis angulospiculatus and Xestospongia muta) were quantified along a shallow to mesophotic depth gradient on Grand Cayman Island. Increases in δ 15 N for all sponges were observed as depth increased, indicating an increasing reliance on heterotrophic food resources. Species-specific changes in symbiotic microbial community composition were also observed as depth increased, and the predicted functional genes associated with nitrogen and carbon cycling showed species-specific changes between depths. Regardless of species-specific changes in microbiome community structure or function, or whether sponges were classified as high microbial or low microbial abundance, sponges increased their consumption of particulate organic matter with increasing depth into the lower mesophotic zone. 
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  2. Sponges in the genus Agelas produce a diversity of bromopyrrole alkaloid secondary metabolites, some of which are known to inhibit predators and pathogens. Selective pressures on sponges to produce chemical defenses vary in time and space, often resulting in differences in the production of secondary metabolites. To characterize intraspecific variation in these compounds, we generated metabolomic profiles of the Caribbean sponge A. tubulata across spatial gradients, including multiple sites in Belize and Grand Cayman, and depths ranging from 15 to 61 m in Grand Cayman. Samples were also analyzed from a reciprocal transplant experiment across shallow (22 m) to mesophotic (61 m) reefs. We found quantitative, but not qualitative, differences in metabolite profiles across sites and depths, with 9 metabolites contributing to that variation. In addition, transplanting sponges across depths resulted in significant changes in concentrations of the metabolite sceptrin. Sponge extracts exhibited antibacterial activity against a panel of marine and human pathogens. Multiple regression analyses showed that different metabolites were associated with antibacterial activity against different pathogens. The strongest compound-specific relationship was a negative effect of oroidin on the growth of Serratia marcescens, and purified oroidin was found to inhibit S. marcescens growth in a dose-dependent manner. Overall, A. tubulata exhibits intraspecific variability in the production of antibacterial secondary metabolites across sites and depths that signals selective responses to its environment. Given the current increase in sponge densities, and incidence of disease on coral reefs, these data have implications for disease resistance and resilience of sponges in the Anthropocene. 
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