Overlooked for a long time, wood falls on the deep oceanic floor are now recognized as extreme, reducing ecosystems based on chemosynthesis, in the same way that hydrothermal vent, cold seeps and whale falls with which they share important physicochemical and faunistic similarities. In these ecosystems, bacterial chemosynthesis using reduced compounds (H2S, CH4) and/or digestion of refractory organic compounds (cellulose and lignin in wood, organic matrix of bone) by heterotrophic bacteria play an essential trophic role, and appear to promote establishment of mutualistic symbioses between bacteria and metazoans for the exploitation of environmental resources. Beyond the description of the taxonomic composition of fauna associated with these sites, current studies aim to understand the functioning and evolutionary links of these ecosystems. Digestive and/or chemoautotrophic associations with microorganisms that could be the key to survive in these habitats have been described in a number of molluscs and annelids but only few crustaceans. The latter distinguish from the other groups by the fact they do not realize intracellular symbioses (endosymbioses) but mainly carry ectosymbioses on their integument (epidermis and cuticle) even in the digestive tract that is partly lined by epidermis (in stomodeum and proctodeum). Moreover, up to now, nothing was known about crustaceans from deep-sea wood falls, their feeding habits and associations with microorganisms. The present work focused on the crustaceans and especially 15 species of decapods from wood falls in the South Pacific Ocean. The specimens were recovered during several French cruises (Salomon2, BOA1, SantoBOA, SalomonBOA3) organized near the Vanuatu, Solomon Island and New Caledonia by the Muséum National d’Histoire Naturelle of Paris. The investigation of the feeding biology and microbial associations of the decapod species combined three complementary approaches : 1) a morphological approach using light and electron microscopy to describe the external and gut structures as well as the gut content and microflora, 2) a trophic approach based on stable isotopes analyses (C and N) together with the nature of the gut content and 3) a molecular approach based on 16S rRNA gene analyses and FISH labeling to identify and locate the bacteria in the gut content (ingested bacteria) and on the gut lining (resident bacteria or symbionts).
Taxonomic identification and morphological observations of the species provided a first non-exhaustive inventory of the best represented decapod species in wood accumulations in the deep South Pacific and point out three interesting informations. 1) Most decapods from the sunken woods belong to Reptantia. 2) Some species (e.g. Munidopsis spp.) belong to deep sea taxa and exhibit typical characteristics while others (e.g. Xylopagurus) much more resemble shallow-water species, indicating that the colonization of wood falls by decapods may have occurred from both deep and coastal habitats. 3) Association degree with sunken wood could be important for some species that appear endemic (e.g. Pylochelidae, Xylopagurus) or which probably have their complete life cycle on wood falls (most of the gravid females having big eggs with a direct larval development).
From a trophic point of view, morphological observations of the digestive system, examinations of gut contents and stable isotope analyses carried out on 15 species, allow us to classify the decapod crustacean from deep-sea sunken woods into two major groups, detritivores and predators/scavengers, and four trophic guilds (trophic levels) depending on two primary food sources, the wood and the particulate organic matrix (“marine snow”). The four guilds were identified as bacteriovorous detritivores (M. nitida, M. pilosa, M. bispinoculata, Munidopsis sp.1), xylophagous detritivores (M. andamanica, R. amboinensis), omnivores (X. caledonicus) and predators/scavengers (pylochelid species, Munida spp, Axiidae sp.1, M. cylindrophthalma). A fifth guild could be represented by limivorous detritivores (C. acutirostella, Alpheidae sp.1). This breaks with the widespread idea that decapod crustaceans from wood falls are all scavengers or predators. In addition, these results have evidenced of some special dietary strategies and highlight the importance of xylophagous decapods in the ecosystem, by their mechanical degradation of the substrate and their production of feces which should enrich the sediment and have a significant impact on the composition of wood and sediment microbiota (bacteria and fungi).
Microscopic observations also allowed us to identify potentially symbiotic associations with resident (and transient) microorganisms (bacteria and/or trichomycetes) in the hindgut of five detritivorous species. Owing that, three of these species have a “wood-based” diet (M. andamanica, M. nitida, R. amboinensis) and considering the location of these microflora, it is strongly suggested that the resident microorganisms are directly related to the xylophagous diet of the host and most particularly involved in the digestion of wood.
More detailed in two xylophagous species, M. andamanica and R. amboinensis, not only these microflora can be distinguished on the basis of their morphology, location and probable genetic differences but also by their roles and involvement in the digestion of the “woody” substrate. The digestive bacteria of M. andamanica could act as trophic intermediates while those of R. amboinensis probably only help in its digestion. From the stable isotopes ratio, it is likely that R. amboinensis directly assimilates the wood digestion products while M. andamanica feeds on wood with a trophic intermediate.
Cloning and sequencing 16S rRNA gene from the gut bacteria of M. andamanica revealed its resident hindgut microflora is largely dominated by two phylotypes (OTUs) of possibly symbiotic bacteria that belong to Firmicutes and Alphaproteobacteria. Surprisingly, these OTUs are very close to gut bacteria isolated from a coastal thalassinid shrimp N. californiensis and from the Chinese crab E. sinensis. The results raise the question of the development and evolution of digestive symbioses in decapods and crustaceans in relation to the diet and/or adaptation to a special habitat, owing that among galatheid crabs the symbiotic relationships appear closer in certain species (M. andamanica) than in others (M. nitida, M. bispinoculata) and that similar differences appear between taxonomic groups. On the other hand, the genetic proximity of bacteria from distant crustacean species raise the question of an eventual co-evolution of linked hosts and symbionts contrasting with the permanent re-acquisition of symbionts from the medium.