The gills and skin microbiota of five pelagic fish species from the Atlantic Ocean.

The gills and skin microbiota and microbiome of wild fish remain far more under-investigated compared to that of farmed fish species, despite that these animal-microbe interactions hold the same ecophysiological roles in both cases. In this study, the gills and skin bacterial microbiota profiles and their presumptive bacterial metabolisms were investigated in five open-sea fishes: bullet tuna (Auxis sp.), common dolphinfish (Coryphaena hippurus), Atlantic little tunny (Euthynnus alletteratus), Atlantic bonito (Sarda sarda) and Atlantic white marlin (Kajikia albida). Gills and skin tissues were collected from two to three individuals per species, from specimens caught by recreational trolling during summer of 2019, and their bacterial 16S rRNA gene diversity was analysed by high-throughput sequencing. The gills bacterial communities among the five species were clearly different but not the skin bacterial microbiota. The dominant operational taxonomic units belonged to the Moraxellaceae, Pseudomonadaceae, Rhodobacteraceae, Staphylococcaceae and Vibrionaceae families. Despite the differences in taxonomic composition, the presumptive bacterial metabolisms between the gills and skin of the five fishes investigated here were ≥ 94% similar and were dominated by basic metabolism, most likely reflecting the continuous exposure of these tissues in the surrounding seawater.

New Eremophilane-Type Sesquiterpenes from the Marine Sediment-Derived Fungus Emericellopsis maritima BC17 and Their Cytotoxic and Antimicrobial Activities.

The fungal strain BC17 was isolated from sediments collected in the intertidal zone of the inner Bay of Cadiz and characterized as Emericellopsis maritima. On the basis of the one strain-many compounds (OSMAC) approach, four new eremophilane-type sesquiterpenes (1-4), together with thirteen known derivatives (5-17) and two reported diketopiperazines (18, 19), were isolated from this strain. The chemical structures and absolute configurations of the new compounds were determined through extensive NMR and HRESIMS spectroscopic studies and ECD calculation. Thirteen of the isolated eremophilanes were examined for cytotoxic and antimicrobial activities. PR toxin (16) exhibited cytotoxic activity against HepG2, MCF-7, A549, A2058, and Mia PaCa-2 human cancer cell lines with IC50 values ranging from 3.75 to 33.44 µM. (+)-Aristolochene (10) exhibited selective activity against the fungal strains Aspergillus fumigatus ATCC46645 and Candida albicans ATCC64124 at 471 µM.

Trophic status of a coastal lagoon - marine harbor system: Potential outwelling rates to the Mesoamerican Barrier Reef southern region.

Eutrophication is still a serious problem in many coastal areas, including the tropics, where river discharges of nutrients is usually high. The ecological stability and ecosystem services of the Mesoamerican Barrier Reef System (MBRS), the world's second-largest coral reef system, suffer a generalized impact by riverine discharge of sediment and organic and inorganic nutrients, which may lead to coastal eutrophication and a coral-macroalgal phase shift. However, few data exist on the MRBS coastal zone status, particularly in Honduras. Here, two in situ sampling campaigns were carried out (May 2017 and January 2018) in the Alvarado Lagoon and Puerto Cortés Bay (Honduras). Measurements included water column nutrients, chlorophyll-a (Chla), particulate organic and inorganic matter and net community metabolism, completed with satellite images analysis. The lagoon and bay environments are ecologically different systems and present different sensitivities to seasonal changes in precipitation as shown by the multivariate analysis. Nonetheless, net community production and respiration rates were neither different spatially, nor seasonally. In addition, both environments were highly eutrophic as shown by the TRIX index. Thus, the Puerto Cortés system represents an important source of dissolved nutrients and particulate matter to the coastal zone. Even though offshore, water quality, based on estimated outwelling rates from the Puerto Cortés system to the coastal waters of the southern MRBS region, improved considerably, concentrations of Chla and nutrients remained higher than those typically measured in non-polluted coral reefs in the Caribbean region and the suggested threshold values. In situ monitoring and assessment of these aspects are crucial to evaluate the ecological functioning of and threats on the MBRS, and elaborate and implement adequate policies for integrated management given its regional and global importance.

Deletion of the Bcnrps1 Gene Increases the Pathogenicity of Botrytis cinerea and Reduces Its Tolerance to the Exogenous Toxic Substances Spermidine and Pyrimethanil.

During the infection of grapevine (Vitis vinifera) by the fungus Botrytis cinerea, the concentration of polyamines, which are toxic substances for the phytopathogen, increases in the grape. Nine NRPS genes have been identified in the genome of B. cinerea, yet the function of five of them remains unknown. For this reason, we have studied the expression of the 9 NRPS genes by RT-qPCR in a medium supplemented with sublethal concentrations of three polyamines (1,3-diaminopropane (1,3-DAP), spermidine (SPD), and spermine (SPM)). Our results show that the presence of polyamines in the culture medium triggered the overexpression of the Bcnrps1 gene in the pathogen. Deleting Bcnrps1 did not affect mycelial growth or adaptation to osmotic stress, and we show that its expression is not essential for the cycle of infection of the B. cinerea. However, mutating the Bcnrps1 gene resulted in overexpression of the Bcnrps6 gene, which encodes for the excretion of siderophores of the coprogen family. Moreover, gene deletion has reduced the tolerance of B. cinerea B05.10 to toxic substances such as the polyamine SPD and the fungicide pyrimethanil, and its virulence has increased. Our findings provide new insights into the function of the Bcnrps1 gene and its involvement in the tolerance of B. cinerea against exogenous toxic compounds.

Seasonal cycles of phytoplankton biomass and primary production in a tropical temporarily open-closed estuarine lagoon - The effect of an extreme climatic event.

Temporarily open-closed estuaries and estuarine lagoons are among the most complex aquatic ecosystems, prone to undergo rapid changes in response to global change and other anthropogenic impacts. Nonetheless, studies on the factors that control annual cycles of phytoplanktonic biomass and primary production in such systems, especially tropical ones, are still scarce. Even less information exists on the effect increasingly frequent extreme climatic events (ECE) might have on their dynamics. For this purpose, we monitored the changes in ecological conditions in the Los Micos estuarine lagoon (Honduras) by sampling monthly during an annual cycle that included several changes in the lagoon's mouth phase and attempted to understand which environmental factors affect phytoplanktonic biomass and primary production. We also evaluated the impact of, and recovery from, a tropical storm ECE. Annual mean net production (Pn), integrated for the euphotic zone, (4.3 ± 2.8 gC m-2 d-1) and Chlorophyll a (27.1 ± 19.1 mg m-3) values in Los Micos place it as one of the more productive estuaries worldwide. The physico-chemical characteristics of the lagoon clearly depended on mouth phase; however, the values of Chla and Pn did not show significant differences between the open and closed phases. The application of distance-based multivariate linear models did not show any clear dominant model being able to explain the observed Chla and Pn patterns. The most parsimonious models included among others, salinity, particulate organic carbon and PO43-, which suggests that primary production is controlled by multiple factors. During the ECE, about 19% of DIN, 91% of DSi and PO43-, 60% of particulate organic carbon and nitrogen, and 86% of Chla were exported to the sea, greatly reducing Pn. However, Chla and Pn values recovered to pre-storm levels within 30 days, indicating that these biological variables are highly resilient in Los Micos Lagoon.

Tidal elevation is the key factor modulating burial rates and composition of organic matter in a coastal wetland with multiple habitats.

This study examines long-term burial rates of organic carbon (OC), organic nitrogen (ON), and total sulphur (TS) in a tidal-dominated coastal wetland with a high spatial heterogeneity and habitat diversity, and long history of human impacts, Cádiz Bay (SW Spain). Using replicate sediment cores, we quantified fluxes of these elements over a transect, extending from the lower saltmarsh (Spartina maritima, ~0.3 m mean sea level, MSL) to the lower intertidal region (Zostera noltei, ~ - 0.7 m MSL). Potential organic matter (OM) sources to the sediment were examined using an extensive dataset on carbon and nitrogen stable isotopes, and C:N molar ratios of primary producers in the region. OC burial rates decreased from the sites below MSL (~80 gC·m-2·y-1) to the lower saltmarsh (~50 gC·m-2·y-1), whereas ON burial rates showed an opposite pattern (~3 gN·m-2·y-1 and ~4 gN·m-2·y-1 observed below and above MSL, respectively). TS burial rates (0.5-46 gS·m-2·y-1) did not show any trend along the sea-land gradient. Hence, (tidal) elevation appeared to be an important determinant of sediment biogeochemical properties, and predictor of OM burial rates. The Bayesian mixing model suggested a well-mixed combination of subtidal and terrestrial/high-marsh OM sources to the surface sediments, with no clear indication of an increased contribution from the particular vegetation species inhabiting the sediments. The indication that there is substantial transport, remineralization and cycling of OM between habitats, suggests diversity may play an important role in maintaining this function, reinforcing the idea that a holistic, catchment-scale view is appropriate for understanding and preserving the long-term burial of OM in coastal wetlands.

Water column dissolved silica concentration limits microphytobenthic primary production in intertidal sediments.

Primary production of microphytobenthos (MPB) contributes significantly to the total production in shallow coastal environments. MPB is a diverse community in which diatoms are usually the main microalgal group. Diatoms require N, P, and other nutrients as with other autotrophs, but in addition require silicate to create their outer cell wall. Therefore, dissolved silica (DSi) might be a potential limiting factor for benthic primary production in areas with reduced freshwater input. To test this hypothesis, a microcosm experiment was conducted using intact sediment cores collected from an intertidal mudflat in the Bay of Cádiz and supplied with increasing concentrations of DSi (0, 5, 10, 25, and 45 µmol · L-1 ). After 7 d of enrichment, we determined chlorophyll a and c (Chl a, c) contents, metabolic rates (Net [Pn ] and Areal Gross [PgA ] Production and Light [RL ] and Dark [RD ] Respiration), as well as fluxes of inorganic nutrients across the sediment-water interface. Chl a and c contents increased significantly with respect to the initial conditions but no differences between treatments were found. Both Pn and PgA showed a saturating-like pattern with silicate concentration, reaching maximum rates at a DSi concentration of 45 µmol · L-1 . The addition of DSi also resulted in an increase of DSi and ammonium uptake by the sediment, which was significantly higher in light than in darkness. Our results clearly show that water column DSi concentrations have a direct impact on benthic primary production, also controlling other related processes such as inorganic nutrient fluxes.

Different Types of Diatom-Derived Extracellular Polymeric Substances Drive Changes in Heterotrophic Bacterial Communities from Intertidal Sediments.

Intertidal areas support extensive diatom-rich biofilms. Such microphytobenthic (MPB) diatoms exude large quantities of extracellular polymeric substances (EPS) comprising polysaccharides, glycoproteins and other biopolymers, which represent a substantial carbon pool. However, degradation rates of different EPS components, and how they shape heterotrophic communities in sediments, are not well understood. An aerobic mudflat-sediment slurry experiment was performed in the dark with two different EPS carbon sources from a diatom-dominated biofilm: colloidal EPS (cEPS) and the more complex hot-bicarbonate-extracted EPS. Degradation rate constants determined over 9 days for three sediment fractions [dissolved organic carbon (DOC), total carbohydrates (TCHO), and (cEPS)] were generally higher in the colloidal-EPS slurries (0.105-0.123 d-1) compared with the hot-bicarbonate-extracted-EPS slurries (0.060-0.096 d-1). Addition of hot-bicarbonate-EPS resulted in large increases in dissolved nitrogen and phosphorous by the end of the experiment, indicating that the more complex EPS is an important source of regenerated inorganic nutrients. Microbial biomass increased ~4-6-fold over 9 days, and pyrosequencing of bacterial 16S rRNA genes revealed that the addition of both types of EPS greatly altered the bacterial community composition (from 0 to 9 days) compared to a control with no added EPS. Bacteroidetes (especially Tenacibaculum) and Verrucomicrobia increased significantly in relative abundance in both the hot-bicarbonate-EPS and colloidal-EPS treatments. These differential effects of EPS fractions on carbon-loss rates, nutrient regeneration and microbial community assembly improve our understanding of coastal-sediment carbon cycling and demonstrate the importance of diverse microbiota in processing this abundant pool of organic carbon.

Dynamics of Inorganic Nutrients in Intertidal Sediments: Porewater, Exchangeable, and Intracellular Pools.

The study of inorganic nutrients dynamics in shallow sediments usually focuses on two main pools: porewater (PW) nutrients and exchangeable (EX) ammonium and phosphate. Recently, it has been found that microphytobenthos (MPB) and other microorganisms can accumulate large amounts of nutrients intracellularly (IC), highlighting the biogeochemical importance of this nutrient pool. Storing nutrients could support the growth of autotrophs when nutrients are not available, and could also provide alternative electron acceptors for dissimilatory processes such as nitrate reduction. Here, we studied the magnitude and relative importance of these three nutrient pools (PW, IC, and EX) and their relation to chlorophylls (used as a proxy for MPB abundance) and organic matter (OM) contents in an intertidal mudflat of Cadiz Bay (Spain). MPB was localized in the first 4 mm of the sediment and showed a clear seasonal pattern; highest chlorophylls content was found during autumn and lowest during spring-summer. The temporal and spatial distribution of nutrients pools and MPB were largely correlated. Ammonium was higher in the IC and EX fractions, representing on average 59 and 37% of the total ammonium pool, respectively. Similarly, phosphate in the IC and EX fractions accounted on average for 40 and 31% of the total phosphate pool, respectively. Nitrate in the PW was low, suggesting low nitrification activity and rapid consumption. Nitrate accumulated in the IC pool during periods of moderate MPB abundance, being up to 66% of the total nitrate pool, whereas it decreased when chlorophyll concentration peaked likely due to a high nitrogen demand. EX-Nitrate accounted for the largest fraction of total sediment nitrate, 66% on average. The distribution of EX-Nitrate was significantly correlated with chlorophyll and OM, which probably indicates a relation of this pool to an increased availability of sites for ionic adsorption. This EX-Nitrate pool could represent an alternative nitrate source with significant concentrations available to the microbial community, deeper in the sediment below the oxic layer.

Oxycline formation induced by Fe(II) oxidation in a water reservoir affected by acid mine drainage modeled using a 2D hydrodynamic and water quality model - CE-QUAL-W2.

The Sancho reservoir is an acid mine drainage (AMD)-contaminated reservoir located in the Huelva province (SW Spain) with a pH close to 3.5. The water is only used for a refrigeration system of a paper mill. The Sancho reservoir is holomictic with one mixing period per year in the winter. During this mixing period, oxygenated water reaches the sediment, while under stratified conditions (the rest of the year) hypoxic conditions develop at the hypolimnion. A CE-QUAL-W2 model was calibrated for the Sancho Reservoir to predict the thermocline and oxycline formation, as well as the salinity, ammonium, nitrate, phosphorous, algal, chlorophyll-a, and iron concentrations. The version 3.7 of the model does not allow simulating the oxidation of Fe(II) in the water column, which limits the oxygen consumption of the organic matter oxidation. However, to evaluate the impact of Fe(II) oxidation on the oxycline formation, Fe(II) has been introduced into the model based on its relationship with labile dissolved organic matter (LDOM). The results show that Fe oxidation is the main factor responsible for the oxygen depletion in the hypolimnion of the Sancho Reservoir. The limiting factors for green algal growth have also been studied. The model predicted that ammonium, nitrate, and phosphate were not limiting factors for green algal growth. Light appeared to be one of the limiting factors for algal growth, while chlorophyll-a and dissolved oxygen concentrations could not be fully described. We hypothesize that dissolved CO2 is one of the limiting nutrients due to losses by the high acidity of the water column. The sensitivity tests carried out support this hypothesis. Two different remediation scenarios have been tested with the calibrated model: 1) an AMD passive treatment plant installed at the river, which removes completely Fe, and 2) different depth water extractions. If no Fe was introduced into the reservoir, water quality would significantly improve in only two years. Deeper extractions (3m above the bottom) would also improve the water quality by decreasing the hypoxic zone. However, extractions at the epilimnion would increase the amount of hypoxic water in the reservoir.

Microbes as Engines of Ecosystem Function: When Does Community Structure Enhance Predictions of Ecosystem Processes?

Microorganisms are vital in mediating the earth's biogeochemical cycles; yet, despite our rapidly increasing ability to explore complex environmental microbial communities, the relationship between microbial community structure and ecosystem processes remains poorly understood. Here, we address a fundamental and unanswered question in microbial ecology: 'When do we need to understand microbial community structure to accurately predict function?' We present a statistical analysis investigating the value of environmental data and microbial community structure independently and in combination for explaining rates of carbon and nitrogen cycling processes within 82 global datasets. Environmental variables were the strongest predictors of process rates but left 44% of variation unexplained on average, suggesting the potential for microbial data to increase model accuracy. Although only 29% of our datasets were significantly improved by adding information on microbial community structure, we observed improvement in models of processes mediated by narrow phylogenetic guilds via functional gene data, and conversely, improvement in models of facultative microbial processes via community diversity metrics. Our results also suggest that microbial diversity can strengthen predictions of respiration rates beyond microbial biomass parameters, as 53% of models were improved by incorporating both sets of predictors compared to 35% by microbial biomass alone. Our analysis represents the first comprehensive analysis of research examining links between microbial community structure and ecosystem function. Taken together, our results indicate that a greater understanding of microbial communities informed by ecological principles may enhance our ability to predict ecosystem process rates relative to assessments based on environmental variables and microbial physiology.

Kinetics of Indigenous Nitrate Reducing Sulfide Oxidizing Activity in Microaerophilic Wastewater Biofilms.

Nitrate decreases sulfide release in wastewater treatment plants (WWTP), but little is known on how it affects the microzonation and kinetics of related microbial processes within the biofilm. The effect of nitrate addition on these properties for sulfate reduction, sulfide oxidation, and oxygen respiration were studied with the use of microelectrodes in microaerophilic wastewater biofilms. Mass balance calaculations and community composition analysis were also performed. At basal WWTP conditions, the biofilm presented a double-layer system. The upper microaerophilic layer (~300 µm) showed low sulfide production (0.31 µmol cm-3 h-1) and oxygen consumption rates (0.01 µmol cm-3 h-1). The anoxic lower layer showed high sulfide production (2.7 µmol cm-3 h-1). Nitrate addition decreased net sulfide production rates, caused by an increase in sulfide oxidation rates (SOR) in the upper layer, rather than an inhibition of sulfate reducing bacteria (SRB). This suggests that the indigenous nitrate reducing-sulfide oxidizing bacteria (NR-SOB) were immediately activated by nitrate. The functional vertical structure of the biofilm changed to a triple-layer system, where the previously upper sulfide-producing layer in the absence of nitrate split into two new layers: 1) an upper sulfide-consuming layer, whose thickness is probably determined by the nitrate penetration depth within the biofilm, and 2) a middle layer producing sulfide at an even higher rate than in the absence of nitrate in some cases. Below these layers, the lower net sulfide-producing layer remained unaffected. Net SOR varied from 0.05 to 0.72 µmol cm-3 h-1 depending on nitrate and sulfate availability. Addition of low nitrate concentrations likely increased sulfate availability within the biofilm and resulted in an increase of both net sulfate reduction and net sulfide oxidation by overcoming sulfate diffusional limitation from the water phase and the strong coupling between SRB and NR-SOB syntrophic relationship.

Nitrate reduction functional genes and nitrate reduction potentials persist in deeper estuarine sediments. Why?

Denitrification and dissimilatory nitrate reduction to ammonium (DNRA) are processes occurring simultaneously under oxygen-limited or anaerobic conditions, where both compete for nitrate and organic carbon. Despite their ecological importance, there has been little investigation of how denitrification and DNRA potentials and related functional genes vary vertically with sediment depth. Nitrate reduction potentials measured in sediment depth profiles along the Colne estuary were in the upper range of nitrate reduction rates reported from other sediments and showed the existence of strong decreasing trends both with increasing depth and along the estuary. Denitrification potential decreased along the estuary, decreasing more rapidly with depth towards the estuary mouth. In contrast, DNRA potential increased along the estuary. Significant decreases in copy numbers of 16S rRNA and nitrate reducing genes were observed along the estuary and from surface to deeper sediments. Both metabolic potentials and functional genes persisted at sediment depths where porewater nitrate was absent. Transport of nitrate by bioturbation, based on macrofauna distributions, could only account for the upper 10 cm depth of sediment. A several fold higher combined freeze-lysable KCl-extractable nitrate pool compared to porewater nitrate was detected. We hypothesised that his could be attributed to intracellular nitrate pools from nitrate accumulating microorganisms like Thioploca or Beggiatoa. However, pyrosequencing analysis did not detect any such organisms, leaving other bacteria, microbenthic algae, or foraminiferans which have also been shown to accumulate nitrate, as possible candidates. The importance and bioavailability of a KCl-extractable nitrate sediment pool remains to be tested. The significant variation in the vertical pattern and abundance of the various nitrate reducing genes phylotypes reasonably suggests differences in their activity throughout the sediment column. This raises interesting questions as to what the alternative metabolic roles for the various nitrate reductases could be, analogous to the alternative metabolic roles found for nitrite reductases.

Photosynthetic activity and community shifts of microphytobenthos covered by green macroalgae.

Macroalgae blooms, a frequent consequence of eutrophication in coastal areas, affect the photosynthetic activity of sediments dominated by microphytobenthos (MPB). Light spectra, steady-state (after 1 h) microprofiles of O2 , gross photosynthesis (Pg ), community respiration in light (RL ) and net community photosynthesis (Pn ) were measured in diatom- and cyanobacteria-dominated communities below increasing layers of Ulva. Photosynthetic photon flux (PPF) decreased exponentially with increasing layers of algae and the light spectrum was increasingly enriched in the green and deprived in blue and red regions. Sediment Pg , Pn and RL decreased as the number of Ulva layers increased; however, 1.6 times higher macroalgal density was necessary to fully inhibit cyanobacteria Pg compared with diatoms, indicating that cyanobacteria were better adapted to this light environment. Long-term (3 weeks) incubations of diatom-dominated sediments below increasing layers of Ulva resulted in a shift in the taxonomic composition of the MPB towards cyanobacteria. Hence, changes in the light climate below macroalgal accumulations can negatively affect the photosynthetic activity of sediments. However, spectral niche differentiation of MPB taxonomic groups and concurrent changes in the MPB community may provide sediments with increased resilience to the detrimental effects of eutrophication.

Changes in benthic denitrification, nitrate ammonification, and anammox process rates and nitrate and nitrite reductase gene abundances along an estuarine nutrient gradient (the Colne estuary, United Kingdom).

Estuarine sediments are the location for significant bacterial removal of anthropogenically derived inorganic nitrogen, in particular nitrate, from the aquatic environment. In this study, rates of benthic denitrification (DN), dissimilatory nitrate reduction to ammonium (DNRA), and anammox (AN) at three sites along a nitrate concentration gradient in the Colne estuary, United Kingdom, were determined, and the numbers of functional genes (narG, napA, nirS, and nrfA) and corresponding transcripts encoding enzymes mediating nitrate reduction were determined by reverse transcription-quantitative PCR. In situ rates of DN and DNRA decreased toward the estuary mouth, with the findings from slurry experiments suggesting that the potential for DNRA increased while the DN potential decreased as nitrate concentrations declined. AN was detected only at the estuary head, accounting for approximately 30% of N2 formation, with 16S rRNA genes from anammox-related bacteria also detected only at this site. Numbers of narG genes declined along the estuary, while napA gene numbers were stable, suggesting that NAP-mediated nitrate reduction remained important at low nitrate concentrations. nirS gene numbers (as indicators of DN) also decreased along the estuary, whereas nrfA (an indicator for DNRA) was detected only at the two uppermost sites. Similarly, nitrate and nitrite reductase gene transcripts were detected only at the top two sites. A regression analysis of log(n + 1) process rate data and log(n + 1) mean gene abundances showed significant relationships between DN and nirS and between DNRA and nrfA. Although these log-log relationships indicate an underlying relationship between the genetic potential for nitrate reduction and the corresponding process activity, fine-scale environmentally induced changes in rates of nitrate reduction are likely to be controlled at cellular and protein levels.