Optical propertiers of the dissolved organic matter as tracers of microbiological and geochemical processes in marine ecosystems

Oceans store 685 Pg of organic carbon of which 662 Pg are in a dissolved form. The diversity of compounds that make up the dissolved organic matter (DOM) pool and the low concentration of each compound make the chemical characterization of this material a difficult task. For that reason, less than 11% of the oceanic DOM has been identified. A variable fraction of the DOM ¿between 20% in the open ocean and 70% in coastal areas- absorbs UV and visible radiation and it is known as coloured DOM (CDOM). A sub-fraction of the CDOM emits the absorbed radiation as fluorescence, although with a low quantum yield (around 1%), and this is called fluorescent DOM (FDOM). The study of the CDOM and FDOM pools, combining the spectroscopy of absorption and fluorescence, allows us to obtain knowledge about (i) the molecular structure of the DOM (i.e., aromaticity and average molecular weigh) and (ii) its biological and photochemical reactivity in a relatively simple, fast and economic way. This can be done through the study of the production, utilization and/or chemical alteration of the different chromophores and fluorophores in response to the activity of the microorganisms and the solar radiation in the ocean. The work that has resulted in this thesis has involved both laboratory experiments and field studies. Some experiments have deepened our knowledge of (or focused on??) the microbiological sources of the CDOM and FDOM. For example, our work has shown that marine phytoplankton produces a fluorophore at Ex/Em 320/410 nm which is consumed by marine bacteria which at the same time produce another absorbing fluorophore at Ex/Em 340/440. These ¿humic-like¿ fluorophores, known in the literature as ¿pico-M¿ and ¿pico-C¿, are considered to be characteristic of marine and continental ecosystems, respectively. This work suggests that differentiation is mostly due to the type of cells that produce them: eukaryotic and prokaryotic cells. Furthermore, DOM isolated by tangential ultrafiltration (> 1 KDa) from different aquatic environments has also been characterized. Significant changes were observed in the aromaticity and average molecular weigh of the samples depending on whether they were of continental or marine origin and also on the exposition to the sunlight before sampling. Moreover, controlled experiments were performed in order to study the response of these materials to natural radiation. These experiments showed degradation of the humic-like fluorophores ¿peak-M¿ and ¿peak-C¿ and the formation of another protein-like fluorophores, known in literature as ¿pico-T¿. When the marine bacteria were cultivated using the irradiated materials as substrate a rapid recovery of the humic-like fluorophores was observed. This recovery was proportional to the initial fluorescence of the materials before irradiation. Finally, we have also studied the relative importance of the processes that involve the mixing between water masses of continental and marine origin, microbial production and photochemical degradation on the CDOM and FDOM distribution of two distinct coastal ecosystem: the ¿Ría de Vigo¿ and the Blanes Bay. The Ría de Vigo, enclosed in the Iberian upwelling system, is periodically affected by downwelling and upwelling events. Microbial production was the dominant process during the donwelling period while the photochemical decomposition predominated during upwellings. On the other hand, Blanes Bay, in the oligotrophic Northwest Mediterranean Sea, possesses a seasonal cycle determined by natural radiation. This is characterized by the accumulation of chromophores and fluorophores absorbing at 300nm during the summer season.Los océanos albergan 685 Pg de carbono orgánico, de los que 662 Pg están en forma disuelta. La enorme diversidad de compuestos que constituyen la materia orgánica disuelta (DOM) y la baja concentración en que se encuentra cada uno de ellos, hace de la caracterización química y estructural de este material una ardua tarea. Es por eso que menos del < 11% de la DOM está identificado en la actualidad. Una fracción variable de la DOM –entre el 20% en océano abierto y el 70% en zonas costeras– absorbe luz UV y visible, por lo que se conoce como DOM coloreada (CDOM). Parte de la CDOM, emite la radiación absorbida en forma de fluorescencia, si bien con un rendimiento cuántico bajo (en torno al 1%) y es conocida como DOM fluorescente (FDOM). El estudio simultaneo de la CDOM y FDOM combinando espectroscopia de absorción y fluorescencia permite –de forma relativamente simple, rápida y barata– ahondar en el conocimiento de (i) la estructura molecular de la DOM, en aspectos tales como su aromaticidad y peso molecular medio; y (ii) su reactividad biológica y fotoquímica, a través del estudio de la producción, consumo y/o alteración química de diferentes grupos cromóforos y fluoróforos en respuesta a la actividad de los microorganismos y la radiación solar en los océanos. En esta Tesis se han realizado tanto experimentos de laboratorio como estudios de campo. En una serie de experimentos se ha profundizado en las fuentes microbiológicas de la CDOM y FDOM en condiciones controladas, demostrando que el fitoplancton marino produce un fluoróforo a Ex/Em = 320 nm/410 nm que es consumido por las bacterias marinas, que a su vez producen otro fluoróforo a Ex/Em = 340 nm/440 nm. Estos fluoróforos de naturaleza húmica, conocidos en la literatura especializada como “pico-M” y “pico-C”, se consideraban característicos de ecosistemas marinos y continentales, respectivamente. Este trabajo sugiere que la diferenciación tiene más que ver con el tipo de células que las producen: eucariotas o procariotas. Se ha caracterizado ópticamente DOM aislada por filtración tangencial (> 1 KDa) de diversas aguas naturales, observándose cambios significativos en la aromaticidad y peso molecular medio de las muestras en función de su origen continental o marino y de su exposición a la luz natural antes de ser colectadas. Igualmente, se realizaron experimentos controlados para estudiar la respuesta de estos materiales a la radiación natural, observándose degradación de los fluoróforos de naturaleza húmica “pico-M” y “pico-C” y generación de un fluoróforo de naturaleza protéica, conocido en la literatura como “pico-T”. Al cultivar bacterias marinas usando los materiales irradiados como substrato se observa una rápida recuperación de los fluoróforos de naturaleza húmica, proporcional a la fluorescencia inicial de los materiales antes de ser irradiados. Finalmente, se ha estudiado la importancia relativa de los procesos de mezcla de masas de agua de origen continental y marino, producción microbiana y degradación fotoquímica sobre la distribución de CDOM y FDOM en dos ecosistemas costeros con distintas condiciones: la Ría de Vigo y la Bahía de Blanes. La Ría de Vigo, sistema eutrófico enclavado en el afloramiento ibérico, se ve afectada periódicamente por episodios de afloramiento y hundimiento, resultando la producción microbiana el proceso dominante en condiciones de afloramiento y la descomposición fotoquímica en condiciones de hundimiento. Por otro lado, la Bahía de Blanes, en el oligotrófico Mediterráneo Nororiental, describe un marcado ciclo estacional dictado por la radiación natural incidente caracterizado por la acumulación estival de cromóforos y fluoróforos que absorben a 300 nm.Els oceans alberguen 685 Pg de carboni orgànic, dels quals 662 Pg estan en forma dissolta. L’enorme diversitat de compostos que constitueixen la matèria orgànica dissolta (DOM) i la baixa concentració en què es troba cadascun d’ells, fa de la caracterització química i estructural d’aquest material una àrdua tasca. És per això que menys del < 11% de la DOM està identificat a dia d’avui. Una fracció variable de la DOM –entre el 20% a l’oceà obert i el 70% a zones costaneres– absorbeix llum UV i visible, per la qual cosa es coneix com DOM acolorida (CDOM). Part de la CDOM, emet la radiació absorbida en forma de fluorescència, si bé amb un rendiment quàntic baix (entorn del 1%) i és coneguda com DOM fluorescent (FDOM). L’estudi simultanei de la CDOM i la FDOM combinant espectroscòpia d’absorció i fluorescència permet –de forma relativament simple, ràpida i barata– aprofundir en el coneixement de (i) l’estructura molecular de la DOM, en aspectes tals com la seva aromaticitat i el pes molecular mitjà; i (ii) la seva reactivitat biològica i fotoquímica, a través de l’estudi de la producció, consum i/o alteració química de diferents grups cromòfors i fluoròfors en resposta a l’activitat dels microorganismes i la radiació solar en els oceans. En aquesta Tesi s’han realitzat tant experiments de laboratori com estudis de camp. En una sèrie d’experiments s’ha aprofundit en les fonts microbiològiques de la CDOM i FDOM en condicions controlades, demostrant que el fitoplàncton marí produeix un fluoròfors a Ex/Em = 320 nm/410 nm que és consumit pels bacteris marins, que al seu torn produeixen un altre fluoròfors a Ex/Em = 340 nm/440 nm. Aquests fluoròfors de naturalesa húmica, coneguts en la literatura especialitzada com “pic-M” i “pic-C”, es consideraven característics d’ecosistemes marins i continentals, respectivament. Aquest treball suggereix que la diferenciació té més a veure amb el tipus de cèl·lules que les produeixen: eucariotes o procariotes. S’ha caracteritzat òpticament DOM aïllada per filtració tangencial (> 1 KDa) de diverses aigües naturals, observant-se canvis significatius en la aromaticitat i pes molecular mitjà de les mostres en funció del seu origen continental o marí i de la seva exposició a la llum natural abans de ser mostrejadas. Igualment, es van realitzar experiments controlats per estudiar la resposta d’aquests materials a la radiació natural, observant-se degradació dels fluoròfors de naturalesa húmica “pic-M” i “pic-C” i generació d’un fluoròfors de naturalesa protéica, conegut en la literatura com “pic-T”. En cultivar bacteris marins utilizant els materials irradiats com a substrat s’observa una ràpida recuperació dels fluoròfors de naturalesa húmica, proporcional a la fluorescència inicial dels materials abans de ser irradiats. Finalment, s’ha estudiat la importància relativa dels processos de barreja de masses d’aigua d’origen continental i marí, producció microbiana i degradació fotoquímica sobre la distribució de CDOM i FDOM en dos ecosistemes costaners diferents: la Ria de Vigo i la Badia de Blanes. La Ria de Vigo, sistema eutròfic enclavat en l’aflorament ibèric, es veu afectada periòdicament per episodis d’aflorament i enfonsament, resultant la producció microbiana el procés dominant en condicions d’aflorament i la descomposició fotoquímica en condicions d’enfonsament. D’altra banda, la Badia de Blanes, en el oligotròfic Mediterrani Nord-oriental, descriu un marcat cicle estacional dictat per la radiació natural incident caracteritzat per l’acumulació estival de cromòfors i fluoròfors que absorbeixen a 300 nm

Dissolved Organic Matter in the Gulf of Cadiz: Distribution and Drivers of Chromophoric and Fluorescent Properties

The Gulf of Cadiz (GoC) connects the Mediterranean Sea with the Atlantic Ocean through the Strait of Gibraltar. Particular hydrographic processes take place in the GoC, such as riverine discharges and surface circulation marked by wind-induced seasonal upwelling. Although physical processes have been widely studied, little is known about the biogeochemical processes that occur in the basin, especially those involving organic matter. Therefore, vertical and seasonal dynamics of dissolved organic carbon (DOC) and optical properties of dissolved organic matter (DOM, absorbance and fluorescence) were measured in 766 samples collected between 5 and 800 m depth during four oceanographic cruises to obtain quantitative and qualitative information about DOM in the GoC. We performed parallel factor analysis (PARAFAC) to identify the main fluorophores present in the GoC, and an optimum multiparameter water mass analysis to differentiate the effect of water mass mixing from the biogeochemical processes in deep waters. PARAFAC analysis validated six fluorescent components; three humic-like, two protein-like, and a possible mixture of polycyclic aromatic hydrocarbon-like with protein-like material. DOC average concentration was 77.0 +/- 12.7 mu M, with higher values in surface and coastal waters during summer, mainly related to primary production. Linear relationships between DOC and apparent oxygen utilization indicate differences in oxygen consumption within the deep waters, which could be related to upwelling zones. Seasonal and spatial differences were also observed in the distribution of fluorescent DOM. Protein-like components were the most abundant fraction, with an average contribution of 64.75% +/- 7.85%, being higher in summer and surface waters, associated with an increase in biological activity. Our results indicate that water mass mixing is the main driver of the major humic-like components, while biogeochemical processes at a local scale explain DOC and protein-like components distribution. Our findings suggest that modeling DOM dynamics in the GoC is complicated due to its complex hydrography and the presence of multiple sources and sinks of DOM

Submarine mud volcanoes as a source of chromophoric dissolved organic matter to the deep waters of the Gulf of Cadiz

Seafloor structures related to the emission of different fluids, such as submarine mud volcanoes (MVs), have been recently reported to largely contribute with dissolved organic matter (DOM) into the oceans. Submarine MVs are common structures in the Gulf of Cadiz. However, little is known about the biogeochemical processes that occur in these peculiar environments, especially those involving DOM. Here, we report DOM characterization in the sediment pore water of three MVs of the Gulf of Cadiz. Estimated benthic fluxes of dissolved organic carbon (DOC) and chromophoric DOM (CDOM) were higher than in other marine sediments with an average of 0.11 +/- 0.04 mmol m(-2) d(-1) for DOC and ranging between 0.11 and 2.86 m(-1) L m(-2) d(-1), for CDOM. Protein-like components represented similar to 70% of the total fluorescent DOM (FDOM). We found that deep fluids migration from MVs (cold seeps) and anaerobic production via sulfate-reducing bacteria represent a source of DOC and FDOM to the overlying water column. Our results also indicate that fluorescent components can have many diverse sources not captured by common classifications. Overall, MVs act as a source of DOC, CDOM, and FDOM to the deep waters of the Gulf of Cadiz, providing energy to the microbial communities living there

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

Research articleMarine dissolved organic matter (DOM) is one of the largest reservoirs of reduced carbon on Earth. In the dark ocean (4200 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 B350 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).Versión del editor10,015

Effect of marine heat waves on carbon metabolism, optical characterization, and bioavailability of dissolved organic carbon in coastal vegetated communities

Dissolved organic carbon (DOC) plays an essential role in the global marine carbon cycle, with coastal vegetated communities as important DOC producers. However, the ultimate fate of this DOC remains still largely unknown due to the lack of knowledge about its chemical composition and lability. Furthermore, global change could alter both DOC fluxes and its bioavailability, affecting the carbon sequestration capacity of coastal vegetated communities. This study explores, in two contrasting seasons (winter and summer), the effects of an in situ simulated marine heatwave on carbon metabolism and DOC fluxes produced by seagrass (Cymodocea nodosa) and macroalgae (Caulerpa prolifera) communities. In addition, the fluorescent characteristics and biodegradability of the dissolved organic matter released directly by the communities under such conditions are evaluated. Under marine heatwave conditions, a significant increase in net community production (NCP) in C. nodosa and a shift to negative DOC fluxes in C. prolifera were observed. In control treatments, the seagrass-dominated community produced a substantial amount of labile (between 44% and 58%) and recalcitrant DOC (between 42% and 56%), while C. prolifera community produced mainly recalcitrant DOC (between 64% and 87%). Therefore, this research revealed that temperature is an important factor determining the NCP in benthic communities and the chemical structure and bioavailability of DOC produced by these communities, since both communities tended to produce more humic-like and less bioavailable DOC with increasing temperature. © 2022 The Authors. Limnology and Oceanography published by Wiley Periodicals LLC on behalf of Association for the Sciences of Limnology and Oceanography

High amino acid osmotrophic incorporation by marine eukaryotic phytoplankton revealed by click-chemistry

pages, 7 figures.-- Data Availability Statement: Raw sequences are publicly available at the European Nucleotide Archive (https://www.ebi.ac.uk/ena) under the accession number PRJEB63614. The datasets generated for this study are available on request to the corresponding authors.The osmotrophic uptake of dissolved organic compounds in the ocean is considered to be dominated by heterotrophic prokaryotes, whereas the role of planktonic eukaryotes is still unclear. We explored the capacity of natural eukaryotic plankton communities to incorporate the synthetic amino acid L-homopropargylglycine (HPG, analogue of methionine) using biorthogonal noncanonical amino acid tagging (BONCAT), and compared it with prokaryotic HPG use throughout a 9-day survey in the NW Mediterranean. BONCAT allows to fluorescently identify translationally active cells, but it has never been applied to natural eukaryotic communities. We found a large diversity of photosynthetic and heterotrophic eukaryotes incorporating HPG into proteins, with dinoflagellates and diatoms showing the highest percentages of BONCAT-labelled cells (49 ± 25% and 52 ± 15%, respectively). Among them, pennate diatoms exhibited higher HPG incorporation in the afternoon than in the morning, whereas small (≤5 μm) photosynthetic eukaryotes and heterotrophic nanoeukaryotes showed the opposite pattern. Centric diatoms (e.g., Chaetoceros, Thalassiosira, Lauderia spp.) dominated the eukaryotic HPG incorporation due to their high abundances and large sizes, accounting for up to 86% of the eukaryotic BONCAT signal, and strongly correlating with bulk 3H-leucine uptake rates. When including prokaryotes, eukaryotes were estimated to account for 19–31% of the bulk BONCAT signal. Our results evidence a large complexity in the osmotrophic uptake of HPG, which varies over time within and across eukaryotic groups, and highlights the potential of BONCAT to quantify osmotrophy and protein synthesis in complex eukaryotic communitiesThis work was funded by the Spanish Ministry of Science, Innovation and Universities (MICINN) through the MIAU (RTI2018-101025-30-B-I00) and MICOLOR (PID2021-125469NB-C31) projects and the Ramon y Cajal contract to C.R.G. (RYC2019-026758-I), with funding from the Spanish Government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S). C.M. was supported by the Juan de la Cierva-formación fellowship (FJC2021-047745-34 I), funded by the Spanish Ministry of Science and Innovation (MCIN/AEI) and the EU (“NextGenerationEU”/PRTR)Peer reviewe

Occurrence and size distribution study of microplastics in household water from different cities in continental Spain and the Canary Islands

9 pages, 5 figures, 1 table, supplementary materials https://doi.org/10.1016/j.watres.2023.120044.-- Data availability: Data will be made available on requestThe purpose of this study was to investigate the occurrence of microplastics (MPs) in drinking water in Spain by comparing tap water from different locations using common sampling and identification procedures. We sampled tap water from 24 points in 8 different locations from continental Spain and the Canary Islands by means of 25 μm opening size steel filters coupled to household connections. All particles were measured and spectroscopically characterized including not only MPs but also particles consisting of natural materials with evidence of industrial processing, such as dyed natural fibres, referred insofar as artificial particles (APs). The average concentration of MPs was 12.5 ± 4.9 MPs/m3 and that of anthropogenic particles 32.2 ± 12.5 APs/m3. The main synthetic polymers detected were polyamide, polyester, and polypropylene, with lower counts of other polymers including the biopolymer poly(lactic acid). Particle size and mass distributions were parameterized by means of power law distributions, which allowed performing estimations of the concentration of smaller particles provided the same scaling parameter of the power law applies. The calculated total mass concentration of the identified MPs was 45.5 ng/L. The observed size distribution of MPs allowed an estimation for the concentration of nanoplastics (< 1 µm) well below the ng/L range; higher concentrations are not consistent with scale invariant fractal fragmentation. Our findings showed that MPs in the drinking water sampled in this work do not represent a significant way of exposure to MPs and would probably pose a negligible risk for human healthThe authors acknowledge the financial support provided by Plastics Europe and that of the Spanish Government, Ministerio de Ciencia e Innovación, grants PID2020-113769RB-C21/C22.With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)Peer reviewe

A nationwide monitoring of atmospheric microplastic deposition

8 Pág.Plastic production continues to increase every year, yet it is widely acknowledged that a significant portion of this material ends up in ecosystems as microplastics (MPs). Among all the environmental compartments affected by MPs, the atmosphere remains the least well-known. Here, we conducted a one-year simultaneous monitoring of atmospheric MPs deposition in ten urban areas, each with different population sizes, economic activities, and climates. The objective was to assess the role of the atmosphere in the fate of MPs by conducting a nationwide quantification of atmospheric MP deposition. To achieve this, we deployed collectors in ten different urban areas across continental Spain and the Canary Islands. We implemented a systematic sampling methodology with rigorous quality control/quality assurance, along with particle-oriented identification and quantification of anthropogenic particle deposition, which included MPs and industrially processed natural fibres. Among the sampled MPs, polyester fibres were the most abundant, followed by acrylic polymers, polypropylene, and alkyd resins. Their equivalent sizes ranged from 22 μm to 398 μm, with a median value of 71 μm. The particle size distribution of MPs showed fewer large particles than expected from a three-dimensional fractal fragmentation pattern, which was attributed to the higher mobility of small particles, especially fibres. The atmospheric deposition rate of MPs ranged from 5.6 to 78.6 MPs m-2 day-1, with the higher values observed in densely populated areas such as Barcelona and Madrid. Additionally, we detected natural polymers, mostly cellulosic fibres with evidence of industrial processing, with a deposition rate ranging from 6.4 to 58.6 particles m-2 day-1. There was a positive correlation was found between the population of the study area and the median of atmospheric MP deposition, supporting the hypothesis that urban areas act as sources of atmospheric MPs. Our study presents a systematic methodology for monitoring atmospheric MP deposition.The authors acknowledge the support provided by Spanish Network of Plastics in the Environment, EnviroPlaNet (www.enviroplanet.net) and the financial support provided by the Spanish Government, Ministerio de Ciencia e Innovación, grants PID2020-113769RB-C21/C22. The authors would like to thank the Interdepartmental Investigation Research Service of the Universidad Autónoma de Madrid (SIDI-UAM and Segainvex) for the use of their infrastructures and their technical support. J.G.S. thanks ACIISI for the contract from the Viera y Clavijo program at the University of La Laguna (85 % co-financed by the European Social Fund).Peer reviewe

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries