Search

Determining accurate rates of benthic nitrogen (N) removal and retention pathways from diverse environments is critical to our understanding of process distribution and constructing reliable N budgets and models. The whole-core N-15 isotope pairing technique (IPT) is one of the most widely used methods to determine rates of benthic nitrate-reducing processes and has provided valuable information on processes and factors controlling N removal and retention in aquatic systems. While the whole core IPT has been employed in a range of environments, a number of methodological and environmental factors may lead to the generation of inaccurate data and are important to acknowledge for those applying the method. In this review, we summarize the current state of the whole core IPT and highlight some of the important steps and considerations when employing the technique. We discuss environmental parameters which can pose issues to the application of the IPT and may lead to experimental artifacts, several of which are of particular importance in environments heavily impacted by eutrophication. Finally, we highlight the advances in the use of the whole-core IPT in combination with other methods, discuss new potential areas of consideration and encourage careful and considered use of the whole-core IPT. With the recognition of potential issues and proper use, the whole-core IPT will undoubtedly continue to develop, improve our understanding of benthic N cycling and allow more reliable budgets and predictions to be made.

This report is related to the BONUS project “Nutrient Cocktails in COAstal zones of the Baltic Sea” alias COCOA. The aim of BONUS COCOA is to investigate physical, biogeochemical and biological processes in a combined and coordinated fashion to improve the understanding of the interaction of these processes on the removal of nutrients along the land-sea interface. The report is especially related to BONUS COCOA WP 6 in which the main objective is extrapolation of results from the BONUS COCOA learning sites to coastal sites around the Baltic Sea in general. Specific objectives of this deliverable (D6.4) were to connect observed process rates with modelling data and ecosystem characteristics. In the report we made statistical analyses of observations from BONUS COCOA study sites together with results from the Swedish Coastal zone Model (SCM). Eight structural variables (water depth, temperature, salinity, bottom water concentrations of oxygen, ammonium, nitrate and phosphate, as well as nitrogen content in sediment) were found common to both the experimentally determined and the model data sets. The observed process rate evaluated in this report was denitrification. In addition regressions were tested between observed denitrification rates and several structural variables (latitude, longitude, depth, light, temperature, salinity, grain class, porosity, loss of ignition, sediment organic carbon, total nitrogen content in the sediment, sediment carbon/nitrogen-ratio, sediment chlorphyll-a as well as bottom water concentrations of oxygen, ammonium, nitrate, and dissolved inorganic phosphorus and silicate) for pooled data from all learning sites. The statistical results showed that experimentally determined multivariate data set from the shallow, illuminated stations was mainly found to be similar to the multivariate data set produced by the SCM model. Generally, no strong correlations of simple relations between observed denitrification and available structural variables were found for data collected from all the learning sites. We found some non-significant correlation between denitrification rates and bottom water dissolved inorganic phosphorous and dissolved silica but the reason behind the correlations is not clear. We also developed and evaluated a theory to relate process rates to monitoring data and nutrient retention. The theoretical analysis included nutrient retention due to denitrification as well as burial of phosphorus and nitrogen. The theory of nutrient retention showed good correlations with model results. It was found that area-specific nitrogen and phosphorus retention capacity in a sub-basin depend much on mean water depth, water residence time, basin area and the mean nutrient concentrations in the active sediment layer and in the water column.

Evidence of aerobic methane oxidation coupled to denitrification has been provided for different freshwater environments, whereas the significance of this process for the marine realm has not been adequately investigated. The goal of this study was to investigate the methane-related reduction of nitrate/nitrite in a marine environment (salinity 8.5). A water sample was collected from the oxic-anoxic transition zone of the Gotland Deep (central Baltic Sea) and the microorganisms contained therein were cultivated in a bioreactor under hypoxic conditions (0.5 μM O2). To enrich the microorganisms involved in the coupled process the bioreactor was continuously sparged with methane as the sole energy and carbon source and simultaneously supplied with a nutrient solution rich in nitrate and nitrite. The bioreactor experiment showed a relationship between the turnover of methane and the concomitant concentration decrease of nitrite and nitrate at the early stage of the experiment. This relationship indicates the role of methanotrophs, which may support heterotrophic denitrifiers by the release of organic compounds as an energy source. Besides, a mixture of uncultured microorganisms, aerobic methanotrophic and heterotrophic denitrifying bacteria were identified in the enrichment culture. Microbial incorporation of nitrite and methane was proven on the cellular and gene levels via 15NO−22-/13CH4 incubation experiments and subsequent analyses with nano secondary ion mass spectrometry (NanoSIMS) and stable isotope probing (SIP). The NanoSIMS showed the incorporation of 15N in almost all the bacteria and in 9% of those there was a concomitant enrichment in 13C. The relatively low abundance of methane-consuming bacteria in the bioreactor was further reflected in specific fatty acids indicative for type I methanotrophic bacteria. Based on pmoA gene analyses, this bacterium is different from the one that was identified as the only key player of methane oxidation in previous studies in the Gotland Deep, indicating the existence of other subordinate methanotrophic bacteria at that site. The results provide the first indications for the predisposition of a methane-related reduction of nitrate/nitrite under hypoxic conditions in the marine realm, supporting the assumption of an interaction between methanotrophic and denitrifying bacteria which hitherto has only been described for fresh water environments.