Hydrogen may be an energy source for endosymbiotic bacteria of the vent mussel Bathymodiolus puteoserpentis

The ultramafic hosted Logatchev hydrothermal vent field at the slow spreading Mid-Atlantic Ridge (MAR) exhibits unusually high hydrogen concentrations due to serpentinization of ultramafic rocks. Endmember H2-concentrations here have been calculated to be as high as 12 mM which is significantly higher than at most other vent sites along the MAR. Hydrogen is a potential energy source for bacteria providing an energy yield of roughly 240 kJ/mol if oxidized with oxygen. Hence, the energy yield is even higher than for conventional aerobic respiration which liberates 220 kJ/mol. The ability to use H2 as an energy source has been shown for a variety of free-living bacteria. However, to date no other energy sources besides methane and sulfide have been identified for vent (or seep) symbionts. Here we show that H2 is consumed by endosymbiotic bacteria of the Logatchev vent mussel Bathymodiolus puteoserpentis. B. puteoserpentis is known to live in dual symbiosis with methane- and sulfide-oxidizing bacteria that occur intracellularly in specialized gill cells called bacteriocytes. The methanotrophic symbionts use methane as both an energy and carbon source whereas the thiotrophic symbionts use H2S as an energy and dissolved CO2 as a carbon source. Hydrothermal fluids carrying methane and sulfide provide the energy for the bacteria and the bacteria in turn provide the mussel with carbon compounds. The mussel on the other hand supplies its symbionts with a constant fluid flow and, by hosting them offers an ideal ecological niche. Freshly dissected gill pieces of B. puteoserpentis incubated in chilled sea water containing hydrogen gas readily consumed H2. The consumption of H2 over time was significantly higher in gill tissues than in symbiont-free mussel tissue indicating that the symbiotic bacteria are responsible for the observed activity. H2-consumption rates were similar in mussels from two different sampling sites, Irina II: 37 nmol h-1 (ml gill)-1 and Quest: 31 nmol h-1 (ml gill)-1. The hydrogen concentrations at these sites did not vary greatly either (Irina II 5.9 μM, Quest 4.2 μM). The H2-oxidation rates decreased significantly after removal of B. puteoserpentis from vent fluids for only 1 day suggesting that hydrogen uptake may be regulated by H2-availability or that bacteria were digested by the host due to starvation. The methane-oxidizing symbiont may be responsible for the observed hydrogen consumption. H2-uptake has been shown for the free-living methanotroph Methylococcus capsulatus and its genes coding for a membrane-bound H2-uptake hydrogenase (hupS and hupL) have been cloned and sequenced. We are currently trying to identify the symbiont responsible for H2-consumption by linking the phylogeny of the symbionts with their physiology using simultaneous fluorescence in situ hybridisation of rRNA and mRNA. Furthermore, we plan to analyze the stable isotope composition of hydrogen in the vent fluids and in the mussels.