Macro- and micro -symbioses involving sponges: Ecological roles in the marine benthos

Abstract

The symbiotic lifestyle represents a fundamental contribution to the diversity of marine ecosystems. Sponges are ideal models to study symbiotic relationships from evolutionary and ecological points of view since they are the most ancient metazoans on Earth, are ubiquitous in the marine benthos, and establish complex symbiosis with both prokaryotes and animals, which in turn harbour their own bacterial communities. In this thesis, we aim to go deeper into the mechanisms by which sponges establish symbiotic associations with members of the three domains of life, combining taxonomical, ecological, and molecular approaches. We study how sponges acquire their symbiotic microbes and whether these microbes contribute to shape the ecological distribution of their hosts. Moreover, we use the sponge-polychaete relationship as an example of multi-partner symbiosis and study the eukaryotic association from the microbial perspective. Finally, we focus on the less studied domain of life, the archaea, to gain insights into the composition and stability of these symbionts in sponges. To assess these goals, we characterized the sponge assemblages in two contrasting environments (well-preserved and impacted) of Nha Trang Bay (Vietnam) and selected the most abundant species for the study of their microbiomes. Additionally, four sponge species harbouring thousands of polychaetes were sampled to analyse the relationships sponge-microbes-polychaetes. Sponges and polychaetes were identified and their respective microbiones and the seawater bacterial communities were analysed by high-throughput sequencing of the 16S rRNA gene (V4 region). We first describe and illustrate the sponges collected to facilitate further taxonomic and faunistic studies in the area. Our samples belonged to 60 species (9 orders, 22 families, and 36 genera) of demosponges. A total of 24 species were added to the already known sponge fauna of Vietnam, from which, 11 species likely represent new species to science. The described species represent an increase of 8 % in the already known sponge list of Vietnam. Our results show that sponge assemblages were more diverse and rich in the well-preserved environments, being dominated by Neofibularia sp. and Aaptos suberitoides in the reefs, and by Monanchora unguiculata, Antho (Antho) sp., and Amphimedon sulcata in rocky habitats. On the other hand, impacted coral reefs were mainly dominated by two abundant species: Clathria reinwardti and Amphimedon paraviridis. Similar ecological metrics were shown by the sponge microbiomes according to the type of habitat, being more diverse in the well-preserved environments. Morever, the sponge microbiomes of the sponge assemblages from the impacted habitats showed higher intra-species dispersion and lower core size (shared ZOTUs across species replicates) than microbiomes of sponges from the well-preserved environments. In this sense, we propose that the Anna Karenina concept, which states that intraspecific variability is higher in dysbiotic than in healthy individuals, can also be applied at the community level for the study sponge assemblages. In our study sponges, bacterial communities were highly stable regardless of the environment, whereas some of their associated polychaetes varied depending on the sampling location. Environmental resilience to different habitat conditions was certainly true for bacterial communities of A. sulcata, the solely species that was found abundant in the two contrasting habitats explored. Moreover, the high overlap in bacteria composition between sponges and seawater suggest microsymbiont acquisition from the environment. In a similar manner, polychaetes were also able to specifically select and enrich some bacteria from their food sponge. Overall, most sequences were shared between biotypes, but at differential abundances, leading to highly specific and stable invertebrate microbiomes, acquired from the environment. Our results support the tenet “Everything is everywhere, but the environment selects”.

La vida en simbiosi representa una contribució fonamental a la diversitat dels ecosistemes marins. Les esponges són models ideals per l’estudi de les relacions simbiòtiques des del punt de vista evolutiu i ecològic, ja que són els metazous més antics de la Terra, són ubiqüistes al bentos marí, i estableixen simbiosis complexes amb procariotes i animals, que al seu torn, contenen les seves pròpies comunitats bacterianes. En aquesta tesis, volem aprofundir en els mecanismes pels quals les esponges estableixen associacions amb membres dels tres dominis de vida, combinant eines taxonòmiques, ecològiques i moleculars. Estudiem com les esponges adquireixen els seus microbis simbionts i com aquests microbis contribueixen a modelar la distribució ecològica de les esponges. A més, utilitzem la relació esponja-poliquet com a exemple de simbiosis multi-organisme i estudiem les associacions eucariotes des de un punt de vista microbià. Finalment, ens centrem en el domini de vida menys estudiat, les arqueas, per aprofundir en la composició i estabilitat d’aquests simbionts en esponges. Per assolir aquests objectius, vam caracteritzar els grups d’esponges de dos ambients diferenciats (impactat i ben preservat) de la badia de Nha Trang (Vietnam), i vam seleccionar les espècies més abundants per l’estudi del seu microbioma. Addicionalment, vam mostrejar quatre espècies d’esponges que contenien milers de poliquets per l’anàlisi de les relacions esponja-microbis-poliquets. Els nostres resultats mostren que les comunitats d’esponges eren més riques i diverses en els ambients ben preservats, i els seus microbiomes mostraven variables ecològiques similars, en els dos tipus d’ambients. La majoria de simbiosis estudiades mostraven una gran especificitat i estabilitat, independentment de l’ambient on vivia l’esponja. El gran solapament entre els bacteris de l’aigua i de l’esponja suggereix que hi ha adquisició microbiana de l’ambient. De forma similar, els poliquets també eren capaços d’adquirir específicament bacteris de les esponges de les quals s’alimentaven. En resum, la majoria de seqüències microbianes eren compartides entre els tres habitats estudiats (aigua/esponge/poliquet), però a diferents abundàncies, donant lloc a microbiomes específics i estables adquirits de l’ambient en els dos grups d’invertebrats estudiats .