Intermediate Levels of Predation and Nutrient Enrichment Enhance the Activity of Ibuprofen-Degrading Bacteria.

Water is the most indispensable natural resource; yet, organic pollution of freshwater sources is widespread. In recent years, there has been increasing concern over the vast array of emerging organic contaminants (EOCs) in the effluent of wastewater treatment plants (WWTPs). Several of these EOCs are degraded within the pore space of riverbeds by active microbial consortia. However, the mechanisms behind this ecosystem service are largely unknown. Here, we report how phosphate concentration and predator-prey interactions drive the capacity of bacteria to process a model EOC (ibuprofen). The presence of phosphate had a significant positive effect on the population growth rate of an ibuprofen-degrading strain. Thus, when phosphate was present, ibuprofen removal efficiency increased. Moreover, low and medium levels of predation, by a ciliated protozoan, stimulated bacterial population growth. This unimodal effect of predation was lost under high phosphate concentration, resulting in the flattening of the relationships between predator density and population growth of ibuprofen degraders. Our results suggest that moderate nutrient and predation levels promote the growth rate of bacterial degraders and, consequently, the self-purifying capability of the system. These findings enhance our understanding of the mechanisms by which riverbed communities drive the processing of EOCs.

The riverine bioreactor: An integrative perspective on biological decomposition of organic matter across riverine habitats.

Riverine ecosystems can be conceptualized as 'bioreactors' (the riverine bioreactor) which retain and decompose a wide range of organic substrates. The metabolic performance of the riverine bioreactor is linked to their community structure, the efficiency of energy transfer along food chains, and complex interactions among biotic and abiotic environmental factors. However, our understanding of the mechanistic functioning and capacity of the riverine bioreactor remains limited. We review the state of knowledge and outline major gaps in the understanding of biotic drivers of organic matter decomposition processes that occur in riverine ecosystems, across habitats, temporal dimensions, and latitudes influenced by climate change. We propose a novel, integrative analytical perspective to assess and predict decomposition processes in riverine ecosystems. We then use this model to analyse data to demonstrate that the size-spectra of a community can be used to predict decomposition rates by analysing an illustrative dataset. This modelling methodology allows comparison of the riverine bioreactor's performance across habitats and at a global scale. Our integrative analytical approach can be applied to advance understanding of the functioning and efficiency of the riverine bioreactor as hotspots of metabolic activity. Application of insights gained from such analyses could inform the development of strategies that promote the functioning of the riverine bioreactor across global ecosystems.

Promoting Individual and Collective Creativity in Science Students.

Creativity is a scientific skill necessary to develop a successful research career. We expose the importance of a growth mindset, divergent, lateral, and associative thinking, serendipity, and being part of a nonhierarchical and diverse research team to improve both individual and collective creativity.

Major imprint of surface plankton on deep ocean prokaryotic structure and activity.

Deep ocean microbial communities rely on the organic carbon produced in the sunlit ocean, yet it remains unknown whether surface processes determine the assembly and function of bathypelagic prokaryotes to a larger extent than deep-sea physicochemical conditions. Here, we explored whether variations in surface phytoplankton assemblages across Atlantic, Pacific and Indian ocean stations can explain structural changes in bathypelagic (ca. 4,000 m) free-living and particle-attached prokaryotic communities (characterized through 16S rRNA gene sequencing), as well as changes in prokaryotic activity and dissolved organic matter (DOM) quality. We show that the spatial structuring of prokaryotic communities in the bathypelagic strongly followed variations in the abundances of surface dinoflagellates and ciliates, as well as gradients in surface primary productivity, but were less influenced by bathypelagic physicochemical conditions. Amino acid-like DOM components in the bathypelagic reflected variations of those components in surface waters, and seemed to control bathypelagic prokaryotic activity. The imprint of surface conditions was more evident in bathypelagic than in shallower mesopelagic (200-1,000 m) communities, suggesting a direct connectivity through fast-sinking particles that escape mesopelagic transformations. Finally, we identified a pool of endemic deep-sea prokaryotic taxa (including potentially chemoautotrophic groups) that appear less connected to surface processes than those bathypelagic taxa with a widespread vertical distribution. Our results suggest that surface planktonic communities shape the spatial structure of the bathypelagic microbiome to a larger extent than the local physicochemical environment, likely through determining the nature of the sinking particles and the associated prokaryotes reaching bathypelagic waters.

Detachment Procedure of Bacteria from Atmospheric Particles for Flow-cytometry Counting.

The protocol separates bacteria from atmospheric particles, obtaining with greater precision their abundance in the atmospheric deposition. This procedure is similar to the one used to separate bacteria in streambed sediments. The detachment procedure consists of a chemical treatment with sodium pyrophosphate and Tween 20 and a physical treatment with agitation and ultrasonic bath to disperse the bacteria in the liquid sample. We recover the total (free and attached) bacteria by generating a density gradient with Nycodenz by centrifugation. The techniques prior to this procedure do not include the microorganisms that are attached to the aerosol particles and, therefore, considerably underestimate the total load and deposition of airborne microorganisms.

Diversity and antimicrobial potential in sea anemone and holothurian microbiomes.

Marine invertebrates, as holobionts, contain symbiotic bacteria that coevolve and develop antimicrobial substances. These symbiotic bacteria are an underexplored source of new bioactive molecules to face the emerging antibiotic resistance in pathogens. Here, we explored the antimicrobial activity of bacteria retrieved from the microbiota of two sea anemones (Anemonia sulcata, Actinia equina) and two holothurians (Holothuria tubulosa, Holothuria forskali). We tested the antimicrobial activity of the isolated bacteria against pathogens with interest for human health, agriculture and aquaculture. We isolated 27 strains with antibacterial activity and 12 of these isolates also showed antifungal activity. We taxonomically identified these strains being Bacillus and Vibrio species the most representative producers of antimicrobial substances. Microbiome species composition of the two sea anemones was similar between them but differed substantially of seawater bacteria. In contrast, microbiome species composition of the two holothurian species was different between them and in comparison with the bacteria in holothurian feces and seawater. In all the holobiont microbiomes Bacteroidetes was the predominant phylum. For each microbiome, we determined diversity and the rank-abundance dominance using five fitted models (null, pre-emption, log-Normal, Zipf and Zipf-Mandelbrot). The models with less evenness (i.e. Zipf and Zipf-Mandelblot) showed the best fits in all the microbiomes. Finally, we tracked (using the V4 hypervariable region of 16S rRNA gene) the relative abundance of these 27 isolates with antibacterial activity in the total pool of sequences obtained for the microbiome of each holobiont. Coincidences, although with extremely low frequencies, were detected only in the microbiome of H. forskali. This fact suggests that these isolated bacteria belong to the long tail of rare symbiotic bacteria. Therefore, more and more sophisticated culture techniques are necessary to explore this apparently vast pool of rare symbiontic bacteria and to determine their biotechnological potentiality.

Sea cucumbers reduce chromophoric dissolved organic matter in aquaculture tanks.

BACKGROUND: Mono-specific aquaculture effluents contain high concentrations of nutrients and organic matter, which affect negatively the water quality of the recipient ecosystems. A fundamental feature of water quality is its transparency. The fraction of dissolved organic matter that absorbs light is named chromophoric dissolved organic matter (CDOM). A sustainable alternative to mono-specific aquaculture is the multitrophic aquaculture that includes species trophically complementary named "extractive" species that uptake the waste byproducts. Sea cucumbers are recognized as efficient extractive species due to the consumption of particulate organic matter (POM). However, the effects of sea cucumbers on CDOM are still unknown. METHODS: During more than one year, we monitored CDOM in two big-volume tanks with different trophic structure. One of the tanks (-holothurian) only contained around 810 individuals of Anemonia sulcata, whereas the other tank (+holothurian) also included 90 individuals of Holothuria tubulosa and Holothuria forskali. We routinely analyzed CDOM absorption spectra and determined quantitative (absorption coefficients at 325 nm) and qualitative (spectral slopes) optical parameters in the inlet waters, within the tanks, and in their corresponding effluents. To confirm the time-series results, we also performed three experiments. Each experiment consisted of two treatments: +holothurians (+H) and -holothurians (-H). We set up three +H tanks with 80 individuals of A. sulcata and 10 individuals of H. tubulosa in each tank and four -H tanks that contained only 80 individuals of A. sulcata. RESULTS: In the time-series, absorption coefficients at 325 nm (a325) and spectral slopes from 275 to 295 nm (S275-295) were significantly lower in the effluent of the +holothurian tank (average: 0.33 m-1 and 16 µm-1, respectively) than in the effluent of the -holothurian tank (average: 0.69 m-1 and 34 µm-1, respectively), the former being similar to those found in the inlet waters (average: 0.32 m-1 and 22 µm-1, respectively). This reduction in the absorption of the dissolved organic matter appears to be mediated by the POM consumption by holothurians. The experiments confirmed the results observed in the time-series. The a325 and S275-295 values were significantly lower in the treatment with holothurians than in the treatment without holothurians indicating a reduction in the concentration of chromophoric organic compounds, particularly of low molecular weight. DISCUSSION: Consequently, sea cucumbers appear to improve water transparency in aquaculture tanks. The underlying mechanism of this improvement might be related to the POM consumption by holothurians, which reduces the concentration of CDOM derived from POM disaggregation or to the direct assimilation of dissolved compounds of low molecular weight as chromophoric amino acids.

Deposition rates of viruses and bacteria above the atmospheric boundary layer.

Aerosolization of soil-dust and organic aggregates in sea spray facilitates the long-range transport of bacteria, and likely viruses across the free atmosphere. Although long-distance transport occurs, there are many uncertainties associated with their deposition rates. Here, we demonstrate that even in pristine environments, above the atmospheric boundary layer, the downward flux of viruses ranged from 0.26 × 109 to >7 × 109 m-2 per day. These deposition rates were 9-461 times greater than the rates for bacteria, which ranged from 0.3 × 107 to >8 × 107 m-2 per day. The highest relative deposition rates for viruses were associated with atmospheric transport from marine rather than terrestrial sources. Deposition rates of bacteria were significantly higher during rain events and Saharan dust intrusions, whereas, rainfall did not significantly influence virus deposition. Virus deposition rates were positively correlated with organic aerosols <0.7 µm, whereas, bacteria were primarily associated with organic aerosols >0.7 µm, implying that viruses could have longer residence times in the atmosphere and, consequently, will be dispersed further. These results provide an explanation for enigmatic observations that viruses with very high genetic identity can be found in very distant and different environments.

Flamingos and drought as drivers of nutrients and microbial dynamics in a saline lake.

Waterbird aggregations and droughts affect nutrient and microbial dynamics in wetlands. We analysed the effects of high densities of flamingos on nutrients and microbial dynamics in a saline lake during a wet and a dry hydrological year, and explored the effects of guano on prokaryotic growth. Concentrations of dissolved organic carbon, total phosphorus and total nitrogen in the surface waters were 2-3 fold higher during the drought and were correlated with salinity. Flamingos stimulated prokaryotic heterotrophic production and triggered cascading effects on prokaryotic abundance, viruses and dissolved nitrogen. This stimulus of heterotrophic prokaryotes was associated with soluble phosphorus inputs from guano, and also from sediments. In the experiments, the specific growth rate and the carrying capacity were almost twice as high after guano addition than in the control treatments, and were coupled with soluble phosphorus assimilation. Flamingo guano was also rich in nitrogen. Dissolved N in lake water lagged behind the abundance of flamingos, but the causes of this lag are unclear. This study demonstrates that intense droughts could lead to increases in total nutrients in wetlands; however, microbial activity is likely constrained by the availability of soluble phosphorus, which appears to be more dependent on the abundance of waterbirds.

Linking optical and molecular signatures of dissolved organic matter in the Mediterranean Sea.

Dissolved organic matter (DOM) plays a key role in global biogeochemical cycles and experiences changes in molecular composition as it undergoes processing. In the semi-closed basins of the oligotrophic Mediterranean Sea, these gradual molecular modifications can be observed in close proximity. In order to extend the spatial resolution of information on DOM molecular composition available from ultrahigh resolution mass spectrometry in this area, we relate this data to optical (fluorescence and absorption spectroscopy) measurements. Covariance between molecular formulae signal intensities and carbon-specific fluorescence intensities was examined by means of Spearman's rank correlations. Fifty two per cent of the assigned molecular formulae were associated with at least one optical parameter, accounting for 70% of the total mass spectrum signal intensity. Furthermore, we obtained significant multiple linear regressions between optical and intensity-weighted molecular indices. The resulting regression equations were used to estimate molecular parameters such as the double bond equivalent, degradation state and occurrence of unsaturated aliphatic compounds from optical measurements. The statistical linkages between DOM molecular and optical properties illustrate that the simple, rapid and cost-efficient optical spectroscopic measurements provide valuable proxy information on the molecular composition of open ocean marine DOM.

Prochlorococcus as a Possible Source for Transparent Exopolymer Particles (TEP).

Transparent exopolymer particles (TEP), usually associated with phytoplankton blooms, promote the formation of marine aggregates. Their exportation to deep waters is considered a key component of the biological carbon pump. Here, we explored the role of solar radiation and picocyanobacteria in the formation of TEP in oligotrophic surface waters of the Atlantic and Pacific Oceans in ten on-deck incubation experiments during the Malaspina 2010 Expedition. TEP concentrations were low on the ocean's surface although these concentrations were significantly higher on the surface of the Pacific (24.45 ± 2.3 µg XG Eq. L-1) than on the surface of the Atlantic Ocean (8.18 ± 4.56 µg XG Eq. L-1). Solar radiation induced a significant production of TEP in the on-deck experiments from the surface water of the Pacific Ocean, reaching values up to 187.3 µg XG Eq. L-1 compared with the low production observed in the dark controls. By contrast, TEP production in the Atlantic Ocean experiments was lower, and its formation was not related to the light treatments. Prochlorococcus sp. from the surface ocean was very sensitive to solar radiation and experienced a high cell decay in the Pacific Ocean experiments. TEP production in the on-deck incubation experiments was closely related to the observed cell decay rates of Prochlorococcus sp., suggesting that this picocyanobacteria genus is a potential source of TEP. The evidence to propose such potential role was derived experimentally, using natural communities including the presence of several species and a variety of processes. Laboratory experiments with cultures of a non-axenic strain of Prochlorococcus marinus were then used to test TEP production by this genus. TEP concentrations in the culture increased with increasing cell abundance during the exponential phase, reaching the highest TEP concentration at the beginning of the stationary phase. The average TEP concentration of 1474 ± 226 µg XG Eq. L-1 (mean ± SE) observed at the stationary phase of P. marinus cultures is comparable with the values reported in the literature for diatom cultures, also growing in non-axenic as well as axenic cultures. Our results identify Prochlorococcus sp. as a possible relevant source of TEP in the oligotrophic ocean.

Turnover time of fluorescent dissolved organic matter in the dark global ocean.

Marine dissolved organic matter (DOM) is one of the largest reservoirs of reduced carbon on Earth. In the dark ocean (>200 m), most of this carbon is refractory DOM. This refractory DOM, largely produced during microbial mineralization of organic matter, includes humic-like substances generated in situ and detectable by fluorescence spectroscopy. Here we show two ubiquitous humic-like fluorophores with turnover times of 435±41 and 610±55 years, which persist significantly longer than the ~350 years that the dark global ocean takes to renew. In parallel, decay of a tyrosine-like fluorophore with a turnover time of 379±103 years is also detected. We propose the use of DOM fluorescence to study the cycling of resistant DOM that is preserved at centennial timescales and could represent a mechanism of carbon sequestration (humic-like fraction) and the decaying DOM injected into the dark global ocean, where it decreases at centennial timescales (tyrosine-like fraction).

Krill excretion boosts microbial activity in the Southern Ocean.

Antarctic krill are known to release large amounts of inorganic and organic nutrients to the water column. Here we test the role of krill excretion of dissolved products in stimulating heterotrophic bacteria on the basis of three experiments where ammonium and organic excretory products released by krill were added to bacterial assemblages, free of grazers. Our results demonstrate that the addition of krill excretion products (but not of ammonium alone), at levels expected in krill swarms, greatly stimulates bacteria resulting in an order-of-magnitude increase in growth and production. Furthermore, they suggest that bacterial growth rate in the Southern Ocean is suppressed well below their potential by resource limitation. Enhanced bacterial activity in the presence of krill, which are major sources of DOC in the Southern Ocean, would further increase recycling processes associated with krill activity, resulting in highly efficient krill-bacterial recycling that should be conducive to stimulating periods of high primary productivity in the Southern Ocean.

Bacterial diversity and composition during rain events with and without Saharan dust influence reaching a high mountain lake in the Alps.

The diversity of airborne microorganisms that potentially reach aquatic ecosystems during rain events is poorly explored. Here, we used a culture-independent approach to characterize bacterial assemblages during rain events with and without Saharan dust influence arriving to a high mountain lake in the Austrian Alps. Bacterial assemblage composition differed significantly between samples with and without Saharan dust influence. Although alpha diversity indices were within the same range in both sample categories, rain events with Atlantic or continental origins were dominated by Betaproteobacteria, whereas those with Saharan dust intrusions were dominated by Gammaproteobacteria. The high diversity and evenness observed in all samples suggests that different sources of bacteria contributed to the airborne assemblage collected at the lake shore. During experiments with bacterial assemblages collected during rain events with Saharan dust influence, cell numbers rapidly increased in sterile lake water from initially ∼3 × 103 cell ml-1 to 3.6-11.1 x105 cells ml-1 within 4-5 days, and initially, rare taxa dominated at the end of the experiment. Our study documents the dispersal of viable bacteria associated to Saharan dust intrusions travelling northwards as far as 47° latitude.

Significance of bacterial activity for the distribution and dynamics of transparent exopolymer particles in the Mediterranean sea.

The study of transparent exopolymer particles (TEP) in the Mediterranean Sea is particularly relevant as they can be promoters of mucilage events, a frequent phenomenon there. We assessed the influence of bacterioplankton on TEP distribution and dynamics across the west-east axis of the Mediterranean Sea. We performed an extensive study of TEP, dissolved carbohydrates, and their relationships with bacterial abundance and bacterial production (BP). A significant and positive relationship was observed between BP and TEP in the study region (r (2) = 0.51, p < 0.001). The direct release of TEP by bacteria was experimentally corroborated using regrowth cultures where increases in TEP tracked bacterial growth in abundance and production. These TEP increases were positively related to the increases in BP (r (2) = 0.78, p < 0.05). The consistency (similar slopes) of the regression lines between BP and TEP in natural conditions and between the increases of BP and TEP in the experiments underlines the relevant role of bacteria in the formation of TEP in this area.

Viability and potential for immigration of airborne bacteria from Africa that reach high mountain lakes in Europe.

We have analysed the diversity of the bacteria, which grow after addition of concentrated airborne particles and desert dust in different microcosms combinations with water samples from oligotrophic alpine lakes. We used, on the one hand, airborne bacteria transported by an African dust plume and collected in a high mountain area in the central Pyrenees (Spain). On the other hand, we collected desert dust in Mauritania (c. 3000 km distance, and a few days estimated airborne journey), a known source region for dust storms in West Africa, which originates many of the dust plumes landing on Europe. In all the dust-amended treatments we consistently observed bacterial growth of common phyla usually found in freshwater ecosystems, i.e. Alpha-, Beta- and Gammaproteobacteria, Actinobacteria, and a few Bacteroidetes, but with different composition based on lake water pretreatment and dust type. Overall, we tentatively split the bacterial community in (i) typical freshwater non-airborne bacteria, (ii) cosmopolitan long-distance airborne bacteria, (iii) non-freshwater low-distance airborne bacteria, (iv) non-freshwater long-distance airborne soil bacteria and (v) freshwater non-soil airborne bacteria. We identified viable long-distance airborne bacteria as immigrants in alpine lakes (e.g. Sphingomonas-like) but also viable putative airborne pathogens with the potential to grow in remote alpine areas (Acinetobacter-like and Arthrobacter-like). Generation of atmospheric aerosols and remote dust deposition is a global process, largely enhanced by perturbations linked to the global change, and high mountain lakes are very convenient worldwide model systems for monitoring global-scale bacterial dispersion and pathogens entries in remote pristine environments.

Spatiotemporal drivers of dissolved organic matter in high alpine lakes: Role of Saharan dust inputs and bacterial activity.

The effects of many environmental stressors such as UV radiation are mediated by dissolved organic matter (DOM) properties. Therefore, determining the factors shaping spatial and temporal patterns is particularly essential in the most susceptible, low dissolved organic carbon (DOC) lakes. We analyzed spatiotemporal variations in dissolved organic carbon concentration and dissolved organic matter optical properties (absorption and fluorescence) in 11 transparent lakes located above tree line in the Sierra Nevada Mountains (Spain), and we assessed potential external (evaporation and atmospheric deposition) and internal (bacterial abundance, bacterial production, chlorophyll a, and catchment vegetation) drivers of DOM patterns. At spatial and temporal scales, bacteria were related to chromophoric DOM (CDOM). At the temporal scale, water soluble organic carbon (WSOC) in dust deposition and evaporation were found to have a significant influence on DOC and CDOM in two Sierra Nevada lakes studied during the ice-free periods of 2000-2002. DOC concentrations and absorption coefficients at 320 nm were strongly correlated over the spatial scale (n = 11, R(2) = 0.86; p < 0.01), but inconsistently correlated over time, indicating seasonal and interannual variability in external factors and a differential response of DOC concentration and CDOM to these factors. At the continental scale, higher mean DOC concentrations and more CDOM in lakes of the Sierra Nevada than in lakes of the Pyrenees and Alps may be due to a combination of more extreme evaporation, and greater atmospheric dust deposition.

Effects of dissolved organic matter photoproducts and mineral nutrient supply on bacterial growth in Mediterranean inland waters.

Sunlight reacts with dissolved organic matter (DOM) modifying its availability as bacterial substrate. We assessed the impact of DOM photoproducts and mineral nutrient supply on bacterial growth in seven inland waters from the South of Spain, where DOM is characterized by low chromophoric content and long residence time. Factorial experiments were performed with presence vs absence of DOM photoproducts and mineral nutrient supply. In six of the seven experiments, we found a significant and negative effect of DOM photoproducts on bacterial growth and a significant and positive effect of mineral nutrient supply. The interaction of these two factors leaded to a compensation of negative effects of photoproducts by availability of mineral nutrients. Dissolved organic matter diagenetic status and the ionic environment where organic carbon is dissolved can be influencing bacterial DOM processing.