Oligo-heterotrophic activity of Marinobacter subterrani creates an indirect Fe(II)-oxidation phenotype in gradient tubes

Autotrophic bacteria utilizing Fe(II) as their energy and electron sources for growth affect multiple biogeochemical cycles. Some chemoheterotrophic bacteria have also been considered to exhibit an Fe(II) oxidation phenotype. For example, several Marinobacter strains have been reported to oxidize Fe(II) based on formation of oxidized iron bands in semi-solid gradient tubes that produce opposing concentration gradients of Fe(II) and oxygen. While gradient tubes are a simple and visually compelling method to test for Fe(II) oxidation, this method alone cannot confirm if, and to what extent, Fe(II) oxidation is linked to metabolism in chemoheterotrophic bacteria. Here we probe the possibility of protein-mediated and metabolic byproduct-mediated Fe(II) oxidation in Marinobacter subterrani JG233, a chemoheterotroph previously proposed to oxidize Fe(II). Results from conditional and mutant studies, along with measurements of Fe(II) oxidation rates suggest M. subterrani is unlikely to facilitate Fe(II) oxidation under microaerobic conditions. We conclude that the Fe(II) oxidation phenotype observed in gradient tubes inoculated with M. subterrani JG233 is a result of oligo-heterotrophic activity, shifting the location where oxygen dependent chemical Fe(II) oxidation occurs, rather than a biologically-mediated process. Importance Gradient tubes are the most commonly used method to isolate and identify neutrophilic Fe(II)-oxidizing bacteria. The formation of oxidized iron bands in gradient tubes provides a compelling assay to ascribe the ability to oxidize Fe(II) to autotrophic bacteria whose growth is dependent on Fe(II) oxidation. However, the physiological significance of Fe(II) oxidation in chemoheterotrophic bacteria is less well understood. Our work suggests that oligo-heterotrophic activity of certain bacteria may create a false-positive phenotype in gradient tubes by altering the location of the abiotic, oxygen-mediated oxidized iron band. Based on the results and analysis presented here, we caution against utilizing gradient tubes as the sole evidence for the capability of a strain to oxidize Fe(II) and that additional experiments are necessary to ascribe this phenotype to new isolates.

Hydrology and riparian forests drive carbon and nitrogen supply and DOC:NO_3⁻ stoichiometry along a headwater Mediterranean stream

In forest headwater streams, metabolic processes are predominately heterotrophic and depend on both the availability of carbon (C) and nitrogen (N) and a favourable C:N stoichiometry. In this context, hydrological conditions and the presence of riparian forests adjacent to streams can play an important, yet understudied role determining dissolved organic carbon (DOC) and nitrate (NO3−) concentrations and DOC:NO3− molar ratios. Here, we aimed to investigate how the interplay between hydrological conditions and riparian forest coverage drives DOC and NO3− supply and DOC:NO3− stoichiometry in an oligotrophic headwater Mediterranean stream. We analysed DOC and NO3− concentrations, and DOC:NO3− molar ratios during both base flow and storm flow conditions at three stream locations along a longitudinal gradient of increased riparian forest coverage. Further, we performed an event analysis to examine the hydroclimatic conditions that favour the transfer of DOC and NO3− from riparian soils to the stream during large storms. Stream DOC and NO3− concentrations were generally low (overall average ± SD was 1.0 ± 0.6 mg C L−1 and 0.20 ± 0.09 mg N L−1), although significantly higher during storm flow compared to base flow conditions in all three stream sites. Optimal DOC:NO3− stoichiometry for stream heterotrophic microorganisms (corresponding to DOC:NO3− molar ratios between 4.8 and 11.7) was prevalent at the midstream and downstream sites under both flow conditions, whereas C-limited conditions were prevalent at the upstream site, which had no surrounding riparian forest. The hydroclimatic analysis of large storm events highlighted different patterns of DOC and NO3− mobilization depending on antecedent soil moisture conditions: drier antecedent conditions promoted rapid elevations of riparian groundwater tables, hydrologically activating a wider and shallower soil layer, and leading to relatively higher increases in stream DOC and NO3− concentrations compared to events preceded by wet conditions. These results suggest that (i) increased supply of limited resources during storms can promote in-stream heterotrophic activity during high flows, especially during large storm events preceded by dry conditions, and (ii) C-limited conditions upstream were gradually overcome downstream, likely due to higher C inputs from riparian forests present at lower elevations. The contrasting spatiotemporal patterns in DOC and NO3− availability and DOC:NO3− stoichiometry observed at the study stream suggests that groundwater inputs from riparian forests are essential for maintaining in-stream heterotrophic activity in oligotrophic, forest headwater catchments.

IMPROVEMENT OF THE METHOD FOR ASSESSMENT OF THE HETEROTROPHIC ACTIVITY OF FRESHWATER BACTERIA BY STUDYING HOW BACTERIAL COMMUNITY CONSUMES THE ORGANIC MATTER EXCRETED BY ALGAE: ISSUES OF WATER QUALITY

The article describes an improved and approved methodology for assessing the heterotrophic activity of freshwater bacteria using a specific example. Namely, the example of studying the bacterial consumption of organic matter excreted by algae. Utilization of organic substances in water bodies by microorganisms and their oxidation are an important part of the functioning of aquatic ecosystems and water self-purification. This article details innovative modifications to the method based on the use of 14C-labeled organic matter by aquatic organisms. All these methods and techniques have been tested in the study of production and destruction processes in freshwater ecosystems of different trophic levels including mesotrophic, eutrophic and hypertrophic surface ecosystems.

Peroxynitrite Generation and Increased Heterotrophic Capacity Are Linked to the Disruption of the Coral–Dinoflagellate Symbiosis in a Scleractinian and Hydrocoral Species

Ocean warming is one of the greatest global threats to coral reef ecosystems; it leads to the disruption of the coral–dinoflagellate symbiosis (bleaching) and to nutrient starvation, because corals mostly rely on autotrophy (i.e., the supply of photosynthates from the dinoflagellate symbionts) for their energy requirements. Although coral bleaching has been well studied, the early warning signs of bleaching, as well as the capacity of corals to shift from autotrophy to heterotrophy, are still under investigation. In this study, we evaluated the bleaching occurrence of the scleractinian coral Mussismillia harttii and the hydrocoral Millepora alcicornis during a natural thermal stress event, under the 2015–2016 El Niño influence in three reef sites of the South Atlantic. We focused on the link between peroxynitrite (ONOO−) generation and coral bleaching, as ONOO− has been very poorly investigated in corals and never during a natural bleaching event. We also investigated the natural trophic plasticity of the two corals through the use of new lipid biomarkers. The results obtained first demonstrate that ONOO− is linked to the onset and intensity of bleaching in both scleractinian corals and hydrocorals. Indeed, ONOO− concentrations were correlated with bleaching intensity, with the highest levels preceding the highest bleaching intensity. The time lag between bleaching and ONOO− peak was, however, species-specific. In addition, we observed that elevated temperatures forced heterotrophy in scleractinian corals, as Mu. harttii presented high heterotrophic activity 15 to 30 days prior bleaching occurrence. On the contrary, a lower heterotrophic activity was monitored for the hydrocoral Mi. alicornis, which also experienced higher bleaching levels compared to Mu. hartii. Overall, we showed that the levels of ONOO− in coral tissue, combined to the heterotrophic capacity, are two good proxies explaining the intensity of coral bleaching.