Microbial community functioning at hypoxic sediments revealed by targeted metagenomics and RNA stable isotope probing

Abstract

Microorganisms are instrumental to the structure and functioning of marine ecosystems and to the chemistry of the ocean due to their essential part in the cycling of the elements and in the recycling of the organic matter. Two of the most critical ocean biogeochemical cycles are those of nitrogen and sulfur, since they can influence the synthesis of nucleic acids and proteins, primary productivity and microbial community structure. Oxygen concentration in marine environments is one of the environmental variables that have been largely affected by anthropogenic activities; its decline induces hypoxic events which affect benthic organisms and fisheries. Hypoxia has been traditionally defined based on the level of oxygen below which most animal life cannot be sustained. Hypoxic conditions impact microbial composition and activity since anaerobic reactions and pathways are favoured, at the expense of the aerobic ones. Naturally occurring hypoxia can be found in areas where water circulation is restricted, such as coastal lagoons, and in areas where oxygen-depleted water is driven into the continental shelf, i.e. coastal upwelling regions. Coastal lagoons are highly dynamic aquatic systems, particularly vulnerable to human activities and susceptible to changes induced by natural events. For the purpose of this PhD project, the lagoonal complex of Amvrakikos Gulf, one of the largest semi-enclosed gulfs in the Mediterranean Sea, was chosen as a study site. Coastal upwelling regions are another type of environment limited in oxygen, where also formation of oxygen minimum zones (OMZs) has been reported. Sediment in upwelling regions is rich in organic matter and bottom water is often depleted of oxygen because of intense heterotrophic respiration. For the purpose of this PhD project, the chosen coastal upwelling system was the Benguela system off Namibia, situated along the coast of south western Africa. The aim of this PhD project was to study the microbial community assemblages of hypoxic ecosystems and to identify a potential link between their identity and function, with a particular emphasis on the microorganisms involved in the nitrogen and sulfur cycles. The methodology that was applied included targeted metagenomics and RNA stable isotope probing (SIP). It has been shown that the microbial community diversity pattern can be differentiated based on habitat type, i.e. between riverine, lagoonal and marine environments. Moreover, the studied habitats were functionally distinctive. Apart from salinity, which was the abiotic variable best correlated with the microbial community pattern, oxygen concentration was highly correlated with the predicted metabolic pattern of the microbial communities. In addition, when the total number of Operational Taxonomic Units (OTUs) was taken into consideration, a negative linear relationship with salinity was identified (see Chapter 2). Microbial community diversity patterns can also be differentiated based on the lagoon under study since each lagoon hosts a different sulfate-reducing microbial (SRM) community, again highly correlated with salinity. Moreover, the majority of environmental terms that characterized the SRM communities were classified to the marine biome, but terms belonging to the freshwater or brackish biomes were also found in stations were a freshwater effect was more evident (see Chapter 3). Taxonomic groups that were expected to be thriving in the sediments of the Benguela coastal upwelling system were absent or present but in very low abundances. Epsilonproteobacteria dominated the anaerobic assimilation of acetate as confirmed by their isotopic enrichment in the SIP experiments. Enhancement of known sulfate-reducers was not achieved under sulfate addition, possibly due to competition for electron donors among nitrate-reducers and sulfate-reducers, to the inability of certain sulfate-reducing bacteria to use acetate as electron donor or to the short duration of the incubations (see Chapter 4). Future research should focus more on the community functioning of such habitats; an increased understanding of the biogeochemical cycles that characterize these hypoxic ecosystems will perhaps allow for predictions regarding the intensity and direction of the cycling of elements, especially of nitrogen and sulfur given their biological importance. Regulation of hypoxic episodes will aid the end-users of these ecosystems to possibly achieve higher productivity, in terms of fish catches, which otherwise is largely compromised by the elevated hydrogen sulfide concentrations.