Heterotrophic Bacterial Production Research Articles

Constant and fluctuating high temperatures interact with Saharan dust leading to contrasting effects on aquatic microbes over time

Mediterranean lakes are facing heightened exposure to multiple stressors, such as intensified Saharan dust deposition, temperature increases and fluctuations linked to heatwaves. However, the combined impact of dust and water temperature on the microbial community in freshwater ecosystems remains underexplored. To assess the interactive effect of dust deposition and temperature on aquatic microbes (heterotrophic bacteria and phytoplankton), a combination of field mesocosm experiments covering a dust gradient (five levels, 0–320 mg L−1), and paired laboratory microcosms with increased temperature at two levels (constant and fluctuating high temperature) were conducted in a high mountain lake in the Spanish Sierra Nevada, at three points in time throughout the ice-free period. Heterotrophic bacterial production (HBP) increased with dust load regardless of the temperature regime. However, temperature regime affected the magnitude and nature of the interactive Dust×T effect on HBP. Specifically, constant and fluctuating high temperature showed opposing interactive effects in the short term that became additive over time. The relationships between HBP and predictor variables (soluble reactive phosphorus (SRP), excreted organic carbon (EOC), and heterotrophic bacterial abundance (HBA)), coupled with an evaluation of the mechanistic variable photosynthetic carbon use efficiency by bacteria (%CUEb), revealed that bacteria depended on primary production in nearly all treatments when dust was added. The %CUEb increased with dust load in the control temperature treatment, but it was highest at intermediate dust loads under both constant and fluctuating high temperatures. Overall, our results suggest that while dust addition alone strengthens algae-bacteria coupling, high temperatures lead to decoupling in the long term at intermediate dust loads, potentially impacting ecosystem function.

Uneven response of phytoplankton-bacteria coupling under Saharan dust pulse and ultraviolet radiation in the south-western Mediterranean Sea

The microbial carbon (C) flux in the ocean is a key functional process governed by the excretion of organic carbon by phytoplankton (EOC) and heterotrophic bacterial carbon demand (BCD). Ultraviolet radiation (UVR) levels in upper mixed layers and increasing atmospheric dust deposition from arid regions may alter the degree of coupling in the phytoplankton-bacteria relationship (measured as BCD:EOC ratio) with consequences for the C-flux through these compartments in marine oligotrophic ecosystem. Firstly, we performed a field study across the south-western (SW) Mediterranean Sea to assess the degree of coupling (BCD:EOC) and how it may be related to metabolic balance (total primary production: community respiration; PPT:CR). Secondly, we conducted a microcosm experiment in two contrasting areas (heterotrophic nearshore and autotrophic open sea) to test the impact of UVR and dust interaction on microbial C flux. In the field study, we found that BCD was not satisfied by EOC (i.e., BCD:EOC >1; uncoupled phytoplankton-bacteria relationship). BCD:EOC ratio was negatively related to PPT:CR ratio across the SW Mediterranean Sea. A spatial pattern emerged, i.e. in autotrophic open sea stations uncoupling was less severe (BCD:EOC ranged 1–2), whereas heterotrophic nearshore stations uncoupling was more severe (BCD:EOC > 2). In the experimental study, in the seawater both enriched with dust and under UVR, BCD:EOC ratio decreased by stimulating autotrophic processes (particulate primary production (PPP) and EOC) in the heterotrophic nearshore area, whereas BCD:EOC increased by stimulating heterotrophic processes [heterotrophic bacterial production (HBP), bacterial growth efficiency (BGE), bacterial respiration (BR)] in the autotrophic open sea. Our results show that this spatial pattern could be reversed under future UVR × Dust scenario. Overall, the impact of greater dust deposition and higher UVR levels will alter the phytoplankton-bacteria C-flux with consequences for the productivity of both communities, their standing stocks, and ultimately, the ecosystem's metabolic balance at the sea surface.

Brownification in the Eastern Mediterranean Sea: effect of simulated terrestrial input on the planktonic microbial food web in an oligotrophic sea

Terrestrial input to marine and freshwater ecosystems colors the water yellow-brown, causing a phenomenon called “brownification”. The effect of brownification on the marine pelagic microbial food web was studied in the oligotrophic eastern Mediterranean in June 2021 by adding HuminFeed in a 15-day mesocosm experiment with 2 treatments: Control (C, no addition) and HuminFeed (HF, single dose of HuminFeed, 2 mg L-1); and 3 replicates per treatment. HuminFeed caused shading, leading to a decrease in the abundance of photo-autotrophic organisms (cyanobacteria Synechococcus and diatoms). Bacteria were positively affected by the HF addition (mainly in terms of production rather than abundance), benefiting either directly from the dissolved organic carbon (DOC) contained in HuminFeed or indirectly from the trophic cascade through the food web. Despite the decrease in HF bacterial abundance during the experiment, an increase in both the high nucleic acid containing bacteria% and heterotrophic bacterial production were observed, suggesting higher activity at the single cell level. In the HF treatment, the increased abundance of dinoflagellates observed could be due to either a dominance of mixotrophic species or a release from predation by copepods. Both ciliates and copepods were severely impacted by HuminFeed, showing lower abundance and distorted forms (ciliates) and reduced reproductive potential (copepods). In conclusion, in the ultraoligotrophic eastern Mediterranean, the simulated brownification negatively affected autotrophs and top predators while benefiting bacteria, thus indicating a shift in the structure of the plankton food web.

Effects on the food-web structure and bioaccumulation patterns of organic contaminants in a climate-altered Bothnian Sea mesocosms

Climate change is expected to alter global temperature and precipitation patterns resulting in complex environmental impacts. The proposed higher precipitation in northern Scandinavia would increase runoff from land, hence increase the inflow of terrestrial dissolved organic matter (tDOM) in coastal regions. This could promote heterotrophic bacterial production and shift the food web structure, by favoring the microbial food web. The altered climate is also expected to affect transport and availability of organic micropollutants (MPs), with downstream effects on exposure and accumulation in biota. This study aimed to assess climate-induced changes in a Bothnian Sea food web structure as well as bioaccumulation patterns of MPs. We performed a mesocosms-study, focusing on aquatic food webs with fish as top predator. Alongside increased temperature, mesocosm treatments included tDOM and MP addition. The tDOM addition affected nutrient availability and boosted both phytoplankton and heterotrophic bacteria in our fairly shallow mesocosms. The increased tDOM further benefitted flagellates, ciliates and mesozooplankton, while the temperature increase and MP addition had minor effect on those organism groups. Temperature, on the other hand, had a negative impact on fish growth and survival, whereas tDOM and MP addition only had minor impact on fish. Moreover, there were indications that bioaccumulation of MPs in fish either increased with tDOM addition or decreased at higher temperatures. If there was an impact on bioaccumulation, moderately lipophilic MPs (log Kow 3.6 – 4.6) were generally affected by tDOM addition and more lipophilic MPs (log Kow 3.8 to 6.4) were generally affected by increased temperature. This study suggest that both increased temperatures and addition of tDOM likely will affect bioaccumulation patterns of MPs in shallow coastal regions, albeit with counteracting effects.

Microbial food web changes induced by terrestrial organic matter and elevated temperature in the coastal northern Baltic Sea

Climate change has been projected to cause increased temperature and amplified inflows of terrestrial organic matter to coastal areas in northern Europe. Consequently, changes at the base of the food web favoring heterotrophic bacteria over phytoplankton are expected, affecting the food web structure. We tested this hypothesis using an outdoor shallow mesocosm system in the northern Baltic Sea in early summer, where the effects of increased temperature (+ 3°C) and terrestrial matter inputs were studied following the system dynamics and conducting grazing experiments. Juvenile perch constituted the highest trophic level in the system, which exerted strong predation on the zooplankton community. Perch subsequently released the microbial food web from heavy grazing by mesozooplankton. Addition of terrestrial matter had a stronger effect on the microbial food web than the temperature increase, because terrestrial organic matter and accompanying nutrients promoted both heterotrophic bacterial production and phytoplankton primary production. Moreover, due to the shallow water column in the experiment, terrestrial matter addition did not reduce the light below the photosynthesis saturation level, and in these conditions, the net-autotrophy was strengthened by terrestrial matter enrichment. In combination with elevated temperature, the terrestrial matter addition effects were intensified, further shifting the size distribution of the microbial food web base from picoplankton to microphytoplankton. These changes up the food web led to increase in the biomass and proportion of large-sized ciliates (>60 µm) and rotifers. Despite the shifts in the microbial food web size structure, grazing experiments suggested that the pathway from picoplankton to nano- and microzooplankton constituted the major energy flow in all treatments. The study implies that the microbial food web compartments in shallow coastal waters will adjust to climate induced increased inputs of terrestrial matter and elevated temperature, and that the major energy path will flow from picoplankton to large-sized ciliates during the summer period.

Distinct Non-conservative Behavior of Dissolved Organic Matter after Mixing Solimões/Negro and Amazon/Tapajós River Waters.

Positive and negative electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry and 1H NMR revealed major compositional and structural changes of dissolved organic matter (DOM) after mixing two sets of river waters in Amazon confluences: the Solimões and Negro Rivers (S + N) and the Amazon and Tapajós Rivers (A + T). We also studied the effects of water mixing ratios and incubation time on the composition and structure of DOM molecules. NMR spectra demonstrated large-scale structural transformations in the case of S + N mixing, with gain of pure and functionalized aliphatic units and loss of all other structures after 1d incubation. A + T mixing resulted in comparatively minor structural alterations, with a major gain of small aliphatic biomolecular binding motifs. Remarkably, structural alterations from mixing to 1d incubation were in essence reversed from 1d to 5d incubation for both S + N and A + T mixing experiments. Heterotrophic bacterial production (HBP) in endmembers S, N, and S + N mixtures remained near 0.03 μgC L-1 h-1, whereas HBP in A, T, and A + T were about five times higher. High rates of dark carbon fixation took place at S + N mixing in particular. In-depth biogeochemical characterization revealed major distinctions between DOM biogeochemical changes and temporal evolution at these key confluence sites within the Amazon basin.

Bacterial transcriptional response to labile exometabolites from photosynthetic picoeukaryote Micromonas commoda

Dissolved primary production released into seawater by marine phytoplankton is a major source of carbon fueling heterotrophic bacterial production in the ocean. The composition of the organic compounds released by healthy phytoplankton is poorly known and difficult to assess with existing chemical methods. Here, expression of transporter and catabolic genes by three model marine bacteria (Ruegeria pomeroyi DSS-3, Stenotrophomonas sp. SKA14, and Polaribacter dokdonensis MED152) was used as a biological sensor of metabolites released from the picoeukaryote Micromonas commoda RCC299. Bacterial expression responses indicated that the three species together recognized 38 picoeukaryote metabolites. This was consistent with the Micromonas expression of genes for starch metabolism and synthesis of peptidoglycan-like intermediates. A comparison of the hypothesized Micromonas exometabolite pool with that of the diatom Thalassiosira pseudonana CCMP1335, analyzed previously with the same biological sensor method, indicated that both phytoplankton released organic acids, nucleosides, and amino acids, but differed in polysaccharide and organic nitrogen release. Future ocean conditions are expected to favor picoeukaryotic phytoplankton over larger-celled microphytoplankton. Results from this study suggest that such a shift could alter the substrate pool available to heterotrophic bacterioplankton.

Compound specific δ15N analysis of amino acids reveals unique sources and differential cycling of high and low molecular weight marine dissolved organic nitrogen

Marine dissolved organic nitrogen is the largest pool of reduced nitrogen (N) in the ocean. Yet, despite indications of a bacterial origin, few N-specific approaches have been developed to trace individual microbial N production and degradation mechanisms. Stable nitrogen isotope ratios (δ15N) are one of the most powerful tools to understand sources and cycling of organic N. However, δ15N measurements of total DON can only be made in the surface ocean and interpretations are complicated by the many factors influencing bulk δ15N values. Compound-specific isotope analysis of amino acids (CSI-AA) on isolated DON fractions represents a new approach for DON study throughout the water column with significantly greater information potential. Here, we compare for the first time δ15N amino acid (δ15N-AA) data on baseline inorganic N source and microbial loop processing in coupled ultrafiltered high molecular weight (HMW) and solid-phase extracted low molecular weight (LMW SPE-) DON from the surface to 2500 m deep in the Atlantic and Pacific Subtropical Gyres.δ15N-AA patterns were very different in HMW versus LMW SPE-DON, indicating unique formation and bacterial alteration mechanisms. HMW DON data supported a prior hypothesis that AA in surface HMW DON originate from heterotrophic bacterial production near the nitracline, but also suggests protozoan heterotrophy as an additional major process. In the mesopelagic, distinct HMW DON δ15N-AA patterns suggest heterotrophic bacterial resynthesis of suspended PON. In contrast, the first LMW DON δ15N-AA patterns were surprisingly similar to those of autotrophs, indicating limited resynthesis at any depth. Taken together with prior biomarker data, we hypothesize a direct bacterial surface source of LMW AA-containing molecules which are inherently refractory. Finally, within each size fraction, δ15N-AA patterns were remarkably similar between Station ALOHA and the BATS site, suggesting our findings can be extended to marine DON at least throughout global subtropical gyres.These findings demonstrate CSI-AA as a novel δ15N tool to understand sources and cycling of marine DON at a new level of detail and suggest a significant shift in our understanding of DON source and cycling. Specifically, we hypothesize that the production and degradation mechanisms of LMW AA-containing DON is fundamentally distinct from HMW DON, and that the majority of LMW refractory DON in the sea derives not from degradation of HMW material, but rather from surface production of intrinsically more refractory LMW molecules.

New Insights for the Renewed Phytoplankton-Bacteria Coupling Concept: the Role of the Trophic Web.

It is widely accepted that in many aquatic ecosystems bacterioplankton is dependent on and regulated by organic carbon supplied by phytoplankton, leading to coupled algae-bacteria relationship. In this study, an in-depth analysis of this relationship has been carried out by combining two approaches: (i) a correlation analyses between heterotrophic bacterial production (BP) vs. primary production (PP) or algal excretion of organic carbon (EOC), (ii) the balance between bacterial carbon demands (BCD) and the supply of C as EOC, measured as BCD:EOC ratio. During the study period (2013-2016), the algae-bacteria relationship was constantly changing from a coupling in 2013, uncoupling in 2014 and 2015, and an incipient return to coupling (in 2016). Our results show that top-down control (bacterivory) by algal mixotrophy acts as a decoupling force since it provides a fresh C source different to algal EOC to satisfy bacterial carbon demands. Notably, a relationship between the BCD:EOC ratio and the ecosystem metabolic balance (Primary production (PP): respiration (R)) was found, suggesting that PP:R may be a good predictor of the algae-bacteria coupling. This analysis, including the comparison between basal and potential ecosystem metabolic balance, can be a tool to improve knowledge on the interaction between both biotics compartments, which the traditional analyses on coupling may not capture.

Glacial Ice Melting Stimulates Heterotrophic Prokaryotes Production on the Getz Ice Shelf in the Amundsen Sea, Antarctica

AbstractExtensive oceanographic data sets were combined with microbiological parameters to elucidate the tight coupling between glacial meltwater and heterotrophic bacterial production (BP) on the Getz Ice Shelf (GtzIS) in the Amundsen Sea. BP in the eastern GtzIS (EG; 85.8 pM Leu. h−1), where basal glacier meltwater upwells, was significantly higher than BP measured in the western GtzIS (WG; 50.6 pM Leu. h−1) and the Amundsen Sea Polynya (ASP; 27.8 pM Leu. h−1). BP in the EG accounted for 49% of primary production, which was greater than that of the WG (10%) and ASP (9.2%). Enhanced BP in the eastern GtzIS was not coupled with phytoplankton biomass, but correlated significantly with the freshwater fraction containing meltwater‐derived dissolved organic carbon (MW‐DOC). These results suggest that warming‐induced glacier melting weakens carbon sequestration efficiency in Antarctic coastal waters by stimulating heterotrophic metabolism that converts MW‐DOC to CO2.

Growth-stage-related shifts in diatom endometabolome composition set the stage for bacterial heterotrophy

Phytoplankton-derived metabolites fuel a large fraction of heterotrophic bacterial production in the global ocean, yet methodological challenges have limited our understanding of the organic molecules transferred between these microbial groups. In an experimental bloom study consisting of three heterotrophic marine bacteria growing together with the diatom Thalassiosira pseudonana, we concurrently measured diatom endometabolites (i.e., potential exometabolite supply) by nuclear magnetic resonance (NMR) spectroscopy and bacterial gene expression (i.e., potential exometabolite uptake) by metatranscriptomic sequencing. Twenty-two diatom endometabolites were annotated, with nine increasing in internal concentration in the late stage of the bloom, eight decreasing, and five showing no variation through the bloom progression. Some metabolite changes could be linked to shifts in diatom gene expression, as well as to shifts in bacterial community composition and their expression of substrate uptake and catabolism genes. Yet an overall low match indicated that endometabolome concentration was not a good predictor of exometabolite availability, and that complex physiological and ecological interactions underlie metabolite exchange. Six diatom endometabolites accumulated to higher concentrations in the bacterial co-cultures compared to axenic cultures, suggesting a bacterial influence on rates of synthesis or release of glutamate, arginine, leucine, 2,3-dihydroxypropane-1-sulfonate, glucose, and glycerol-3-phosphate. Better understanding of phytoplankton metabolite production, release, and transfer to assembled bacterial communities is key to untangling this nearly invisible yet pivotal step in ocean carbon cycling.

Impact of dust addition on the metabolism of Mediterranean plankton communities and carbon export under present and future conditions of pH and temperature

Abstract. Although atmospheric dust fluxes from arid as well as human-impacted areas represent a significant source of nutrients to surface waters of the Mediterranean Sea, studies focusing on the evolution of the metabolic balance of the plankton community following a dust deposition event are scarce, and none were conducted in the context of projected future levels of temperature and pH. Moreover, most of the experiments took place in coastal areas. In the framework of the PEACETIME project, three dust-addition perturbation experiments were conducted in 300 L tanks filled with surface seawater collected in the Tyrrhenian Sea (TYR), Ionian Sea (ION) and Algerian basin (FAST) on board the R/V Pourquoi Pas? in late spring 2017. For each experiment, six tanks were used to follow the evolution of chemical and biological stocks, biological activity and particle export. The impacts of a dust deposition event simulated at their surface were followed under present environmental conditions and under a realistic climate change scenario for 2100 (ca. +3 ∘C and −0.3 pH units). The tested waters were all typical of stratified oligotrophic conditions encountered in the open Mediterranean Sea at this period of the year, with low rates of primary production and a metabolic balance towards net heterotrophy. The release of nutrients after dust seeding had very contrasting impacts on the metabolism of the communities, depending on the station investigated. At TYR, the release of new nutrients was followed by a negative impact on both particulate and dissolved 14C-based production rates, while heterotrophic bacterial production strongly increased, driving the community to an even more heterotrophic state. At ION and FAST, the efficiency of organic matter export due to mineral/organic aggregation processes was lower than at TYR and likely related to a lower quantity/age of dissolved organic matter present at the time of the seeding and a smaller production of DOM following dust addition. This was also reflected by lower initial concentrations in transparent exopolymer particles (TEPs) and a lower increase in TEP concentrations following the dust addition, as compared to TYR. At ION and FAST, both the autotrophic and heterotrophic community benefited from dust addition, with a stronger relative increase in autotrophic processes observed at FAST. Our study showed that the potential positive impact of dust deposition on primary production depends on the initial composition and metabolic state of the investigated community. This impact is constrained by the quantity of nutrients added in order to sustain both the fast response of heterotrophic prokaryotes and the delayed one of primary producers. Finally, under future environmental conditions, heterotrophic metabolism was overall more impacted than primary production, with the consequence that all integrated net community production rates decreased with no detectable impact on carbon export, therefore reducing the capacity of surface waters to sequester anthropogenic CO2.

Spatial and vertical distribution of aerobic and anaerobic dark inorganic carbon fixation in coastal tropical lake sediments

The process of chemosynthesis can represent an important source of organic matter and energy across the benthic zone. However, its detailed vertical distribution and importance in relation to other microbial processes are still poorly investigated in inland aquatic ecosystems. Here, we determined the rates of dark inorganic carbon fixation (DCF) compared to heterotrophic bacterial production (BP) and oxygen consumption (SOC) across a depth profile of oxic to anoxic sediments in the littoral zone of nine tropical lakes located in one of the most important coastal conservation areas, the Jurubatiba Sandbank National Park (Rio de Janeiro, Brazil). Rates of DCF were highly variable (means of 0.1–1.2 mmol C m−2 d−1) among the studied lakes, with mostly production occurring below 10 mm of sediment in the anoxic zone. Although presenting a relatively low contribution in relation to BP and SOC, our study showed extensive microbial metabolism in the anaerobic sediment below the top centimeter, highlighting the importance of studying microbial processes across depth at fine scales. To our knowledge, it is the first study to show detailed vertical DCF rates in coastal tropical lake sediments, thus contributing to a better comprehension of the microbial role in the biogeochemical cycles of aquatic systems.

Response of oligotrophic coastal microbial populations in the SE Mediterranean Sea to crude oil pollution; lessons from mesocosm studies

Anthropogenically-induced oil spills release large amounts of organic pollutants into the marine environment. To date, little is known about the response of microbial populations (biomass, activity and diversity) to crude oil pollution in Low Nutrients Low Chlorophyll and warm systems. Here, we investigated the daily dynamics of phytoplankton and heterotrophic bacteria in response to an oil spill (500 μm thick layer) in the coastal waters of the SE Mediterranean Sea (SEMS), using mesocosms during winter and summer. Crude oil addition caused a marked decrease in phytoplankton biomass (40–76%) and production rates (22–96%), whereas heterotrophic bacterial abundance and production increased (4–68% and 17–165%, respectively). Concurrently, amplicon sequencing of the 16S rRNA gene revealed that oil-degrading bacteria became abundant 48–96 h post-oil addition, while the cosmopolitan Synechococcus and SAR11 lineages were significantly reduced (by 78–98% and 59–98%, respectively). Fertilization with inorganic nutrients (NO3 and PO4) reduced the deleterious effects of the oil, resulting in a less distinct reduction in phytoplankton biomass/abundance. Our results highlight the potential of intrinsic microbial communities to degrade oil-derived pollutants in oligotrophic coastal waters.

Seasonal microbial food web dynamics in contrasting Southern Ocean productivity regimes

AbstractSpatial and seasonal dynamics of microbial loop fluxes were investigated in contrasting productivity regimes in the Indian sector of the Southern Ocean. Observations carried out in late summer (February–March 2018; project MOBYDICK) revealed higher microbial biomasses and fluxes in the naturally iron‐fertilized surface waters of Kerguelen island in comparison to surrounding off‐plateau waters. Differences were most pronounced for bacterial heterotrophic production (2.3‐fold), the abundance of heterotrophic nanoflagellates (HNF; 2.7‐fold). Independent of site, grazing by HNF was the main loss process of bacterial production (80–100%), while virus‐induced mortality was low (< 9%). Combining these results with observations from previous investigations during early spring and summer allowed us to describe seasonal patterns in microbial food web fluxes and to compare these to carbon export in the iron‐fertilized and high‐nutrient, low‐chlorophyll (HNLC) Southern Ocean. Our data suggest an overall less efficient microbial food web during spring and summer, when respiration and viral lysis, respectively, represent important loss terms of bacterially‐mediated carbon. In late summer, primary production is more efficiently transferred to bacterial biomass and HNF and thus available for higher trophic levels. These results provide a new insight into the seasonal variability and the quantitative importance of microbial food web processes for the fate of primary production in the Southern Ocean.

Interaction among bacterioplankton and macrophytes in shallow lakes with high macrophyte cover

Growth of submerged and emergent macrophytes was studied together with heterotrophic bacterioplankton abundance and production in two Hungarian shallow lakes with dominant macrophyte covers. It was expected that bacterioplankton numbers and activity would have an effect on macrophyte biomass accumulation. Bacterial production and abundance showed a strong seasonal pattern with maximum in the warmest months (July, August). It was found that macrophyte biomass increased with heterotrophic bacterial production and abundance up to 5.6 µg C l− 1 h− 1 and 5.30*106 cells, respectively, while over that value was negatively associated with macrophyte growth. It was also shown that the relationship between heterotrophic bacteria and macrophytes also varied seasonally, showing a multifaceted relationship. It was demonstrated that macrophytes are not only the most significant carbon and energy source for the bacteria in shallow, macrophyte-dominated lakes, but are also competing organisms that could be supressed by excessive bacterial activity. These findings could help better understand the interaction between macrophytes and bacterioplankton, and assist wetland managers in quantifying what may be a primary cause of reed die-back.

Bacterial production prevails over photo- and chemosynthesis in a eutrophic tropical lagoon

Dark carbon fixation is a combination of processes performed by microorganisms to fix inorganic carbon driven the energy generated by chemical reactions. Our study quantified microbial primary and secondary production rates to evaluate the relative importance of dark carbon fixation in relation to photosynthetic rates and heterotrophic bacterial production and their contribution to the carbon cycle in a tropical eutrophic and human-impacted lagoon (Rodrigo de Freitas Lagoon, Brazil). Samples were collected daily at a fixed point to assess short-term temporal variations and at six different stations to assess spatial variability. Water samples were incubated using 14C-bicarbonate at 100% of light and in the dark to quantify photosynthesis and dark carbon fixation, respectively; and 3H-leucine incubations were performed to measure bacterial production. Environmental parameters (temperature, salinity, dissolved oxygen, dissolved organic carbon and inorganic nutrients) were measured to characterize the hydrology of the studied area, and to evaluate their influence on the microbial production rates. There were no significant changes in production rates over time or within different depths. Bacterial production rates varied significantly over space, and were correlated with nitrogen compounds, temperature, salinity and pH. However, a short rainfall event, although not strong or extreme, promoted rapid changes in the shallow water column, such as a decrease in temperature and nitrogen compounds, inducing responses to microbial production rates, such as higher rates of dark carbon fixation and bacterial production at the bottom and lower rates of photosynthesis. Heterotrophic bacterial production was significantly higher than dark and light carbon fixation, being the most important carbon-cycling process in Rodrigo de Freitas Lagoon, possibly favored by the high allochthonous organic matter and nutrient inputs. This study showed that, on average, dark carbon fixation accounted for 30% at the surface and 43% near the bottom of the total primary production and up to 10% of heterotrophic production in the water column, which may be related to the availability of ammonia and methane. We concluded that, even for this short-term temporal and spatial assessment, dark carbon fixation is relevant, although usually disregarded, and this process should be taken into account when evaluating carbon budgets, metabolic balance and ecosystem functioning in tropical coastal lagoons.

Dissolved Organic Carbon Source Influences Tropical Coastal Heterotrophic Bacterioplankton Response to Experimental Warming.

Global change impacts on marine biogeochemistry will be partly mediated by heterotrophic bacteria. Besides ocean warming, future environmental changes have been suggested to affect the quantity and quality of organic matter available for bacterial growth. However, it is yet to be determined in what way warming and changing substrate conditions will impact marine heterotrophic bacteria activity. Using short-term (4 days) experiments conducted at three temperatures (−3°C, in situ, +3°C) we assessed the temperature dependence of bacterial cycling of marine surface water used as a control and three different dissolved organic carbon (DOC) substrates (glucose, seagrass, and mangrove) in tropical coastal waters of the Great Barrier Reef, Australia. Our study shows that DOC source had the largest effect on the measured bacterial response, but this response was amplified by increasing temperature. We specifically demonstrate that (1) extracellular enzymatic activity and DOC consumption increased with warming, (2) this enhanced DOC consumption did not result in increased biomass production, since the increases in respiration were larger than for bacterial growth with warming, and (3) different DOC bioavailability affected the magnitude of the microbial community response to warming. We suggest that in coastal tropical waters, the magnitude of heterotrophic bacterial productivity and enzyme activity response to warming will depend partly on the DOC source bioavailability.

Seasonal freshening of NW Mediterranean surface water impacts microbial heterotrophic activity and dissolved organic matter

The Rhone river represents the most important source of freshwater, nutrients and organic matter to the northwestern (NW) Mediterranean Sea and riverine input markedly affects biogeochemistry and ecosystem functioning in the estuarine and coastal zone. Structures of low salinity waters (LSW) originating near the river plume can also be transported along the continental shelf and offshore. The objective of the present study was to investigate the influence of LSW distant from their source, focusing on dissolved organic matter (DOM) and related microbial processes during two annual cycles (2007 and 2008) at a time series site characterized by the regular occurrence of LSW in spring (Microbial Observatory Laboratoire Arago). We observed enhanced bacterial heterotrophic production and community respiration and specific DOM features within these LSW, concurrently with low net community production. Our results suggest that LSW represent a mechanism of labile DOM supply, thereby sustaining enhanced heterotrophic microbial metabolism.

Relevance of Nutrient-Limited Phytoplankton Production and Its Bacterial Remineralization for Carbon and Oxygen Fluxes in the Baltic Sea

The Baltic Sea is prone to oxygen deficiency due to the restricted water exchange with the North Sea in coincidence with a high biological oxygen demand. The partitioning of organic carbon between respiration, accumulation and export is co-determined by phytoplankton primary production and its subsequent bacterial remineralization. Here, we investigated net phytoplankton primary production, heterotrophic bacterial biomass production and dark CO2 fixation by on-board incubations with radiolabeled tracers in the Baltic Proper and in the Gulf of Riga after the main spring bloom. Results show that low phytoplankton standing stocks of ≤1.6 µg chlorphyll a L-1 sustained net primary production of 161-724 mg C m-2 d-1 under nitrogen limitation. Estimates of bacterial carbon remineralization suggest that freshly produced organic carbon was supplied to the aphotic zone at all stations. In the southern Baltic Proper, net primary production exceeded the bacterial carbon demand in the upper mixed layer, suggesting that organic matter derived from nutrient-limited primary production was available for export to bacterial communities below the oxycline. On average, 46% of heterotrophic bacterial production was mediated in oxygen minimum zones, revealing the high importance of organic matter recycling under hypoxic and anoxic conditions for the carbon budget. Dark CO2 fixation of up to 4.33 µg C L-1 d-1 in sulfide-free waters equaled 9-54% of the co-inciding heterotrophic bacterial carbon demand and may have provided another organic carbon source for heterotrophic activity. Substantially higher dark CO2 fixation up to 25.46 µg C L-1 d-1 was determined in sulfidic waters. Since our study was conducted five months after the major Baltic inflow event in winter 2014/2015, potential effects of deep water ventilation could be investigated. In the Gotland Basin, heterotrophic bacterial production in renewed oxygen-rich bottom water was similar to that in the uplifted oxygen-deficient former bottom water, while it was significantly reduced in sulfidic waters. Hence, our results suggest that the removal of hydrogen sufide by inflow events has a high potential to increase bacterial carbon remineralization.