Additional file 1 of Microbiome changes through the ontogeny of the marine sponge Crambe crambe

Additional file 1: Figure S1. Number of shared ASVs between adult individuals and their own brooded larvae from the three individuals analysed. The Upset plot shows inclusive intersections between analysed pairs. Pairs originating from the same individual are marked in the same colour in the matrix. Set size corresponds to core community values (present in all the replicates from the same individual) and bars represent the size (num. ASVs) of the indicated interaction in the matrix ordered by decreasing values. Venn diagrams show specific comparisons between adults-brooded larvae pairs from the same individual

Embryonic features of <i>Mycale acerata</i>.

<p>A. General view of inner blastomeres (bl) of the embryo, containing abundant lipid droplets (li). B. Detail of nucleated (n) blastomere showing lipid droplets (li). C. Multimembrane inclusions (mm) in the blastomeres (bl) of the embryo. D. Detail of lipid droplet. E. Embryonic cell similar to nurse cell (nc) among the embryo blastomeres (bl). F. Detail of rod-shape inclusions (y) in the blastomeres.</p

SEM micrographs of embryos of <i>Mycale acerata</i> and embryos and larvae of <i>Mycale laevis</i>.

<p>A. Two mid-stage embryos (e) of <i>M</i>. <i>acerata</i> brooded within the sponge mesohyl. B. Detached late-stage embryo of <i>M</i>. <i>acerata</i> showing small micromeres (white arrows). C. Nurse cells (nc) of <i>M</i>. <i>acerata</i> lying within the collagenous follicle (white arrows). Inset showing rod-shaped bacteria (white arrow) lying on the collagen layer surrounding the embryo of <i>M</i>. <i>acerata</i>. D. Larva of <i>M</i>. <i>laevis</i> showing the anterior pole (ap), mid part (mp), and posterior pole (pp). Note the spicules (s) occurring around the larva. E. Posterior bare pole (white arrow) of the larva of <i>M</i>. <i>laevis</i> surrounded by ciliated cells (black arrow). Note the numerous spherulous cells (sc) in contact with the collagenous layer surrounding the larva. F. Ciliated cells (white arrow) of the mid part of the larva of <i>M</i>. <i>laevis</i>.</p

Morphological gross features of the embryos of <i>Mycale acerata</i> and embryos and larvae of <i>Mycale laevis</i>.

<p>A. Mid-stage embryo of <i>M</i>. <i>acerata</i>, detached from the tissue, showing macromeres (ma), micromeres (mi), and the follicle (f) surrounding the embryo. B. Late-stage embryo of <i>M</i>. <i>acerata</i>, detached from the tissue. Note the similar size of all blastomeres and the follicle (f) surrounding the embryos. C. Late-stage embryo of <i>M</i>. <i>laevis</i> within the tissue, showing macromeres (ma), micromeres (mi), and the follicle (f) surrounding the embryo. D. Larva of <i>M</i>. <i>laevis</i> within the tissue. Note the follicle (f) of the embryo. Inset showing a larva of <i>M</i>. <i>laevis</i> with large epithelial ciliated cells (ci) oriented in the anterior (a)-posterior (p) axis.</p

Live specimens of <i>Mycale</i> in the study areas.

<p><b>A.</b> Massive specimens of living <i>Mycale acerata</i> in Deception Island, Antarctica. B. Adult specimen of <i>Mycale laevis</i> in Bocas del Toro, Panama. C. Embryos (arrows) brooded in the mesohyl of <i>M</i>. <i>acerata</i>. D. Embryos (white arrows) and ciliated larvae with a bare posterior pole (black arrow) within the mesohyl of <i>M</i>. <i>laevis</i>.</p

Reproductive elements of <i>Mycale acerata</i>.

<p>A. Degenerated oocyte in the mesohyl (m) showing lipid droplets (li) and areas with degenerated cytoplasm (dg). B. Cellular (f) and collagen follicle (co) enveloping embryo with round blastomeres (bl) and large lipid inclusions (li). Note the nurse cell (nc) within the lumen of the follicle. C. Nurse cell (nc) in the vicinity of the follicle (f) that contains the embryo (e). Collagen (co) surrounding the follicle. Note the lipid droplets (li) putatively secreted by nurse cells within the embryo. D. Nucleated (n) nurse cell (nc) transporting large lipid inclusions (li) to the embryo blastomeres (bl). Note the collagen (co) layer surrounding the follicle (f). E. Pinacocyte (pi) layer limiting the aquiferous canal (ca) and collagen layer (co) laying underneath. Mesohyl containing symbiotic bacteria (b) and archaeocyte-like cells (a). F. Detail of mesohyl bacteria (b) within collagen (co) accumulations.</p

Morphological features of all larvae described in the genus <i>Mycale</i>.

<p>Abbreviations: ap = anterior pole, pp = posterior pole</p><p>* = only embryos observed</p><p>? = unknown, All = totally ciliated;- = except.</p><p>Morphological features of all larvae described in the genus <i>Mycale</i>.</p

Larval features in <i>Mycale laevis</i>.

<p>A. Larval cells filled with homogeneous (ho) and heterogeneous (he) yolk, large mitochondria (mi), round large nucleus (n), interspersed bacteria (b), and collagen (co). B. Detail of bacteria (b) among larval cells. C. Detail of morphology of round larval cells with homogeneous (ho) and heterogeneous (he) yolk. D. Detail of heterogeneous yolk platelet. E. Sclerocyte showing a cross-section of siliceous spicule (s) in the mid part of the larva. F. Detail of collagen fibers in the mid part of the larva.</p

Ultrastructure of the embryo and larva of <i>Mycale laevis</i>.

<p>A. Cellular follicle (f) and collagen layer (co) surrounding the embryo. Note the presence of a nurse cell (nc) within the lumen of the embryonic follicle and symbiotic bacteria (b) lying in the collagen layer. B. Posterior end of the larva showing ciliated cells (ci) prior to the bare area, and lipid droplets (li) in the cells. Note that nurse (nc) and spherulous cells (sc) have crossed the follicle (f) and are found within the lumen. C. Round nucleolated (nu) cells in the mid part of the larva showing homogeneous (ho) and heterogeneous (he) yolk, and Golgi apparatus (g). Note the round nucleus of the cells (n). D. Close up of nucleated (n) larval cell showing homogeneous (ho) and heterogeneous (he) yolk. E. Detail of nurse cell outside the embryo showing large rough endoplasmic reticulum (rer), non-nucleolated nucleus (n), several mitochondria (mi), and many homogenous yolk platelets (ho). Note the bacteria (b) in the mesohyl. F. Larval ciliated (ci) cells in the posterior end showing large lipid droplets (li).</p

Summary of the Antarctic and Subantarctic bones analyzed during this study.

<p>Summary of the Antarctic and Subantarctic bones analyzed during this study.</p